Neoantigen Peptide Mimics

Abstract

Disclosed herein are polypeptide fragments and polynucleotides based on mutant capicua transcriptional repressor (CIC), catenin beta 1 (CTNNB1), v-erb-b2 erythroblastic leukemia viral oncogene homolog B (ERBB2), kirsten rat sarcoma (KRAS), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), phosphatase and tensin homolog (PTEN), splicing factor 3b subunit 1 (SF3B1), SRY-box transcription factor 17 (SOX17), tumor protein 53 (TP53), and cytomegalovirus (CMV) sequences, vectors, host cells, viruses, methods for generating CD8+ T-cells, and methods of treatment. Also disclosed herein are T-cell receptors (TCRs), polynucleotides, vectors and cells comprising the TCRs, and methods of treatment.

Description

TECHNICAL FIELD

Provided are polypeptide fragments and polynucleotides based on mutant capicua transcriptional repressor (CIC), catenin beta 1 (CTNNB1), v-erb-b2 erythroblastic leukemia viral oncogene homolog B (ERBB2), kirsten rat sarcoma (KRAS), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), phosphatase and tensin homolog (PTEN), splicing factor 3b subunit 1 (SF3B1), SRY-box transcription factor 17 (SOX17), tumor protein 53 (TP53), and cytomegalovirus (CMV), as well as vectors, host cells, viruses, methods for generating CD8+ T-cells, and methods of treatment. Also provided are T-cell receptors (TCRs), polynucleotides and vectors that encode the TCRs, cells comprising the TCRs, and methods of treatment.

BACKGROUND

Clinical evidence demonstrates the central role for neoantigen-specific T cell responses in cancer. For example, neoantigen load is associated with better clinical outcomes, neoantigen-specific T cells have shown clinical evidence of anti-tumor activity, and neoantigen-specific T cells kill tumor cell in vitro and in vivo. However, recurrent oncogenic mutations are expected to be poor binders to class I HLA alleles.

SUMMARY

Described herein are capicua transcriptional repressor (CIC) polypeptide fragments comprising: an arginine to tryptophan amino acid substitution at a position corresponding to position 215 of SEQ ID NO: 102 (R215W), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02:01. In some embodiments, the CIC polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, the R215W substitution is at amino acid position 8 of the fragment. In further embodiments, the CIC polypeptide fragment is selected from the group consisting of SEQ ID NO: 25, SEQ ID NO: 26 and SEQ ID NO: 27.

Described herein are catenin beta 1 (CTNNB1) polypeptide fragments comprising: (i) a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C), or (i) a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02:01. In certain embodiments, the CTNNB1 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In some embodiments, the S33C substitution is at amino acid position 4 of the fragment. In further embodiments, the S37F substitution is at amino acid position 8 of the fragment. In still further embodiments, the CTNNB1 polypeptide fragment is selected from the group consisting of SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 80, and SEQ ID NO: 81.

Described herein are v-erb-b2 erythroblastic leukemia viral oncogene homolog B (ERBB2) polypeptide fragments comprising a valine to isoleucine amino acid substitution at a position corresponding to position 842 of SEQ ID NO: 104 (V842I), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02:01. In some embodiments, the ERBB2 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, the V8421 substitution at amino acid position 3 of the fragment. In still further embodiments, the ERBB2 polypeptide fragment is selected from the group consisting of SEQ ID NO: 84, SEQ ID NO: 85, and SEQ ID NO: 86.

Described herein are kirsten rat sarcoma (KRAS) polypeptide fragments comprising: (i) a glycine to alanine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12A), (ii) a glycine to cysteine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12C), or (iii) a glycine to valine amino acid substitution at a position corresponding to at position 12 of SEQ ID NO: 105 (G12V), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02:01. In some embodiments, the KRAS polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, the G12A substitution is at amino acid position 7 of the fragment. In further embodiments, the G12C substitution is at amino acid position 7 of the fragment. In still further embodiments, the G12V substitution is at amino acid position 7 of the fragment. In some embodiments, the KRAS polypeptide fragment is selected from the group consisting of SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, and SEQ ID NO: 42.

Described herein are phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) polypeptide fragments comprising: (i) a glutamic acid to lysine amino acid substitution at a position corresponding to position 453 of SEQ ID NO: 106 (E453K), or (ii) a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02:01. In some embodiments, the PIK3CA polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, the E453K substitution is at amino acid position 3 of the fragment. In further embodiments, the G118D substitution is at amino acid position 7 of the fragment. In still further embodiments, the PIK3CA polypeptide fragment is selected from the group consisting of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, and SEQ ID NO: 47.

Described herein are phosphatase and tensin homolog (PTEN) polypeptide fragments comprising: an arginine to cysteine amino acid substitution at a position corresponding to position 173 of SEQ ID NO: 107 (R173C), and an amino acid substitution at amino acid position 3 of the fragment, amino acid position 10 of the fragment, or both, wherein the fragment is ten amino acids in length, and wherein the fragment binds to HLA-A*02:01. In some embodiments, the PTEN polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, the R173C substitution is at amino acid position 1 of the fragment. In further embodiments, the PTEN polypeptide fragment is selected from the group consisting of SEQ ID NO: 48, SEQ ID NO: 49, and SEQ ID NO: 88.

Described herein are splicing factor 3b subunit 1 (SF3B1) polypeptide fragments comprising: an arginine to histidine amino acid substitution at a position corresponding to position 625 of SEQ ID NO: 108 (R625H), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02:01. In certain embodiments, the SF3B1 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native epitope. In some embodiments, the R625H substitution is at amino acid position 7 of the fragment. In further embodiments, the SF3B1 polypeptide fragment is selected from the group consisting of SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 90, SEQ ID NO: 91 and SEQ ID NO: 92.

Described herein are SRY-box transcription factor 17 (SOX17) polypeptide fragments comprising: a serine to isoleucine amino acid substitution at a position corresponding to position 403 of SEQ ID NO: 109 (S403I), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02:01. In certain embodiments, the SOX17 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In some embodiments, the S403I substitution is at amino acid position 6 of the fragment. In further embodiments, the SOX17 polypeptide fragment is selected from the group consisting of SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, and SEQ ID NO: 93.

Described herein are tumor protein 53 (TP53) polypeptide fragments comprising: (i) an arginine to leucine amino acid substitution at a position corresponding to position 110 of SEQ ID NO: 110 (R110L), (ii) a serine to phenylalanine amino acid substitution at a position corresponding to position 127 of SEQ ID NO: 110 (S127F), (iii) a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N), (iv) a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y), (v) a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L), (vi) a histidine to leucine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193L), (vii) a histidine to tyrosine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193Y), (viii) a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C), or (ix) a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02:01. In certain embodiments, the TP53 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In some embodiments, the R110L substitution is at amino acid position 8 of the fragment. In further embodiments, the S127F substitution is at amino acid position 7 of the fragment. In still further embodiments, the K132N substitution is at amino acid position 4 of the fragment. In certain embodiments, the C141Y substitution is at amino acid position 3 of the fragment. In further embodiments, the P152L substitution is at amino acid position 9 of the fragment. In still further embodiments, the H193L substitution is at amino acid position 7 of the fragment. In some embodiments, the Y220C substitution is at amino acid position 4 of the fragment. In further embodiments, the V272M substitution is at amino acid position 9 of the fragment. In still further embodiments, the TP53 polypeptide fragment is selected from the group consisting of SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 94, SEQ ID NO: 95, and SEQ ID NO: 96.

Described herein is a polypeptide fragment selected from the group consisting of SEQ ID NO: 25, SEQ ID NO: 26 and SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, and SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81; SEQ ID NO: 84, SEQ ID NO: 85, and SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 91,SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, and SEQ ID NO: 96.

Described herein is a polypeptide fragment selected from the group consisting of SEQ ID NO: 29, SEQ ID NO: 32, SEQ ID NO: 45, SEQ ID NO: 59, SEQ ID NO: 64, SEQ ID NO: 68, SEQ ID NO: 75 and SEQ ID NO: 78.

Described herein are polynucleotides encoding at least one or more polypeptide fragments provided herein. In certain embodiments, the polynucleotide is cDNA.

Described herein are vectors comprising one or more polynucleotides provided herein. In certain embodiments, the vector is selected from an adenovirus vector, an alphaviral vector, a poxvirus vector, an adeno-associated virus vector, a retrovirus vector, a self-replicating RNA molecule, and a combination thereof. In certain embodiments, the adenovirus vector is selected from hAd5, hAd7, hAd11, hAd26, hAd34, hAd35, hAd48, hAd49, hAd50, GAd20, Gad19, GAd21, GAd25, GAd26, GAd27, GAd28, GAd29, GAd30, GAd31, ChAd3, ChAd4, ChAd5, ChAd6, ChAd7, ChAd8, ChAd9, ChAd10, ChAd11, ChAd16, ChAdI7, ChAd19, ChAd20, ChAd22, ChAd24, ChAd26, ChAd30, ChAd31, ChAd37, ChAd38, ChAd44, ChAd55, ChAd63, ChAd73, ChAd82, ChAd83, ChAd146, ChAd147, PanAd1, PanAd2, and PanAd3. In some embodiments, the poxvirus vector is selected from smallpox virus vector, vaccinia virus vector, cowpox virus vector, monkeypox virus vector, Copenhagen vaccinia virus (W) vector, New York Attenuated Vaccinia Virus (NYVAC) vector, and Modified Vaccinia Ankara (MVA) vector.

Described herein are pharmaceutical compositions comprising at least one or more polypeptide fragments provided herein.

Described herein are pharmaceutical compositions comprising at least one or more polynucleotides provided herein.

Described herein are pharmaceutical compositions comprising at least one or more vectors provided herein.

Described herein are methods of treating cancer in a subject comprising administering to the subject in need thereof the polypeptide fragments, the polynucleotides encoding the polypeptide fragments, the vectors comprising the polynucleotides, or the pharmaceutical compositions described herein.

Described herein are methods of inducing an immune response in a subject comprising administering to the subject in need thereof the polypeptide fragments, the polynucleotides encoding the polypeptide fragments, the vectors comprising the polynucleotides, or the pharmaceutical compositions described herein.

Described herein are methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of CTNNB1 mutant comprising a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 2, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 29, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a catenin beta 1 (CTNNB1) mutant comprising a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 3, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 32, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) mutant comprising a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 9, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 45, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 13, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 59, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 16, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 64, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 18, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 68, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 22, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 75, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 23, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 78, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

In certain embodiments the methods of treatment comprise administering the polynucleotide in part a) prior to administering the polynucleotide in part b). In certain embodiments the methods of treatment comprise administering the polynucleotide in part b) prior to administering the polynucleotide in part a). In certain embodiments the methods of treatment comprise administering the polynucleotide in part a) concurrently with the polynucleotide in part b).

In certain embodiments the methods of treatment comprise administering a vector encoding the polynucleotide of part a) and a vector encoding the polynucleotide of part b). In some embodiments, the vectors are independently selected from an adenovirus vector, an alphaviral vector, a poxvirus vector, an adeno-associated virus vector, a retrovirus vector, and a self-replicating RNA molecule. In further embodiments, the adenovirus vector is selected from hAd5, hAd7, hAd11, hAd26, hAd34, hAd35, hAd48, hAd49, hAd50, GAd20, Gad19, GAd21, GAd25, GAd26, GAd27, GAd28, GAd29, GAd30, GAd31, ChAd3, ChAd4, ChAd5, ChAd6, ChAd7, ChAd8, ChAd9, ChAd10, ChAd11, ChAd16, ChAdI7, ChAd19, ChAd20, ChAd22, ChAd24, ChAd26, ChAd30, ChAd31, ChAd37, ChAd38, ChAd44, ChAd55, ChAd63, ChAd73, ChAd82, ChAd83, ChAd146, ChAd147, PanAd1, PanAd2, and PanAd3. In further embodiments, the poxvirus vector is selected from smallpox virus vector, vaccinia virus vector, cowpox virus vector, monkeypox virus vector, Copenhagen vaccinia virus (W) vector, New York Attenuated Vaccinia Virus (NYVAC) vector, and Modified Vaccinia Ankara (MVA) vector.

Described herein are kits of parts comprising a pair of polypeptide fragments selected from the group consisting of: (a) SEQ ID NO: 2 and SEQ ID NO: 29; (b) SEQ ID NO: 3 and SEQ ID NO: 32; (c) SEQ ID NO: 9 and SEQ ID NO: 45; (d) SEQ ID NO: 13 and SEQ ID NO: 59; (e) SEQ ID NO: 16 and SEQ ID NO: 64; (f) SEQ ID NO: 18 and SEQ ID NO 68; (g) SEQ ID NO: 22 and SEQ ID NO: 75; and (h) SEQ ID NO: 23 and SEQ ID NO: 78.

Described herein are kits of parts comprising a pair of polypeptide fragments selected from the group consisting of: (a) SEQ ID NO: 9 and SEQ ID NO: 45; (b) SEQ ID NO: 13 and SEQ ID NO: 59; and (c) SEQ ID NO: 18 and SEQ ID NO 68.

Described herein are methods for generating CD8+ T-cells that are positive for an HLA-A*02:01-restricted polypeptide fragment and a cognate native polypeptide fragment, comprising exposing CD8+ T-cells to the HLA-A*02:01-restricted polypeptide fragment and cognate native polypeptide fragment selected from the group consisting of: (a) SEQ ID NO: 2 and SEQ ID NO: 29; (b) SEQ ID NO: 3 and SEQ ID NO: 32; (c) SEQ ID NO: 9 and SEQ ID NO: 45; (d) SEQ ID NO: 13 and SEQ ID NO: 59; (e) SEQ ID NO: 16 and SEQ ID NO: 64; (f) SEQ ID NO: 18 and SEQ ID NO 68; (g) SEQ ID NO: 22 and SEQ ID NO: 75; and (h) SEQ ID NO: 23 and SEQ ID NO: 78; and selecting CD8+ T cells that are positive to both the HLA-A*02:01-restricted polypeptide fragment and a cognate neoantigen polypeptide fragment.

Described herein are methods for generating CD8+ T-cells that are positive for an HLA-A*02:01-restricted polypeptide fragment and a cognate native polypeptide fragment, comprising exposing CD8+ T-cells to the HLA-A*02:01-restricted polypeptide fragment and cognate native polypeptide fragment selected from the group consisting of: (a) SEQ ID NO: 9 and SEQ ID NO: 45; (b) SEQ ID NO: 13 and SEQ ID NO: 59; and (c) SEQ ID NO: 18 and SEQ ID NO 68; and selecting CD8+ T cells that are positive to both the HLA-A*02:01-restricted polypeptide fragment and a cognate neoantigen polypeptide fragment.

Described herein are T-cell receptors (TCRs) comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a complementarity determining region 3 (CDR3) comprising an amino acid sequence provided in Table 14, Table 15, Table 16, Table 17, or Table 18, and (b) the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18. An alpha chain CDR3 corresponds to a beta chain CDR3 if they appear in the same row in Table 14, Table 15, Table 16, Table 17, or Table 18.

Described herein are T-cell receptors (TCRs) comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a complementarity determining region 1 (CDR1) comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, a complementarity determining region 2 (CDR2) comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, and a complementarity determining region 3 (CDR3) comprising an amino acid sequence provided in Table 14, Table 15, Table 16, Table 17, or Table 18, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, a CDR2 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, and a CDR3 comprising a corresponding amino acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18. An alpha chain CDR1 or CDR2 corresponds to a beta chain CDR1 or CDR2 if they appear in the same row in Table 19, Table 20, Table 21, Table 22, or Table 23. An alpha chain CDR3 corresponds to a beta chain CDR3 if they appear in the same row in Table 14, Table 15, Table 16, Table 17, or Table 18. An alpha and beta chain CDR1 and CDR2 provided in Table 19, Table 20, Table 21, Table 22, or Table 23 correspond to an alpha and beta chain CDR3 provided in the same row in Table 14, Table 15, Table 16, Table 17 or Table 18.

Described herein are polynucleotides encoding the TCRs provided herein.

Described herein are vectors comprising the polynucleotides provided herein.

Described herein are cells transformed to express the polynucleotides provided herein.

Described herein are cells comprising the vectors provided herein. In certain embodiments, the cell is a CD8+ T cell.

Described herein are pharmaceutical compositions comprising the TCRs, polynucleotides, the vectors, or the cells provided herein.

Described herein are methods of treating cancer in a subject comprising administering to the subject in need thereof a pharmaceutical composition comprising a TCR described herein.

Described herein are methods of inducing an immune response in a subject comprising administering to the subject in need thereof a pharmaceutical composition comprising a TCR described herein.

Described herein are methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) mutant comprising a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D) in a subject comprising administering to the subject in need thereof a TCRs described herein.

Described herein are methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y) in a subject comprising administering to the subject in need thereof a TCR described herein.

Described herein are methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L) in a subject comprising administering to the subject in need thereof a TCR described herein.

BRIEF DESCRIPTION OF THE DRAWINGS

The foregoing summary, as well as the following detailed description of preferred embodiments of the present application, will be better understood when read in conjunction with the appended drawings. It should be understood, however, that the application is not limited to the precise embodiments shown in the drawings.

FIG. 1 illustrates exemplary FACS plots used to determine the frequency of dual positive T cells for tetramers specific for mutant and mimic peptides represented by SEQ ID NO: 9 and SEQ ID NO: 45. The donor used was Lot #19054445 from Hemacare. The gating strategy is mentioned in the figure. Lymphocytes derived from the donor were gated to remove dead cells. Live cells were further gated to remove doublets and other cell aggregates. Finally, CD8+ T cells were gated and the frequency of cells staining positive for both mutant and mimic peptide loaded tetramers (PE+/APC+) was determined. mi_mu refers to a sample in which both mimic and mutant tetramers specific to the peptide were used for staining.

FIG. 2 illustrates exemplary FACS plots used to determine the frequency of dual positive T cells for tetramers specific for mutant and mimic peptides represented by SEQ ID NO: 13 and SEQ ID NO: 59. The donor used was Lot #19054445 from Hemacare. The gating strategy is mentioned in the figure. Lymphocytes derived from the donor were gated to remove dead cells. Live cells were further gated to remove doublets and other cell aggregates. Finally, CD8+ T cells were gated and the frequency of cells staining positive for both mutant and mimic peptide loaded tetramers (PE+/APC+) was determined. mi mu refers to a sample in which both mimic and mutant tetramers specific to the peptide were used for staining.

FIG. 3 illustrates exemplary FACS plots used to determine the frequency of dual positive T cells for tetramers specific for mutant and mimic peptides represented by SEQ ID NO: 18 and SEQ ID NO 68. The donor used was Lot #19054445 from Hemacare. The gating strategy is mentioned in the figure. Lymphocytes derived from the donor were gated to remove dead cells. Live cells were further gated to remove doublets and other cell aggregates. Finally, CD8+ T cells were gated and the frequency of cells staining positive for both mutant and mimic peptide loaded tetramers (PE+/APC+) was determined. mi mu refers to a sample in which both mimic and mutant tetramers specific to the peptide were used for staining.

FIG. 4 illustrates exemplary FACS plots used to determine the frequency of T cells staining positive for negative tetramer on APC fluorescence channel. The negative tetramer is loaded with a non-specific peptide with no known reactivity. The negative tetramer was used as a control to gate on the cells. The donor used was Lot #19054445 from Hemacare. The gating strategy is mentioned in the figure. Lymphocytes derived from the donor were gated to remove dead cells. Live cells were further gated to remove doublets and other cell aggregates. Finally, CD8+ T cells were gated to exclude background signal arising from the negative tetramer. Neg APC refers to a sample in which negative tetramer for APC was used for staining.

FIG. 5 illustrates exemplary FACS plots used to determine the frequency of T cells staining positive for negative tetramer on PE fluorescence channel. The negative tetramer is loaded with a non-specific peptide with no known reactivity. The negative tetramer was used as a control to gate on the cells. The donor used was Lot #19054445 from Hemacare. The gating strategy is mentioned in the figure. Lymphocytes derived from the donor were gated to remove dead cells. Live cells were further gated to remove doublets and other cell aggregates. Finally, CD8+ T cells were gated to exclude background signal arising from the negative tetramer.Neg PE refers to a sample in which negative tetramer for PE was used for staining.

FIG. 6 illustrates exemplary FACS plots used to determine the frequency of dual positive T cells for tetramers specific for mutant and mimic peptides represented by SEQ ID NO: 2 and SEQ ID NO: 29. The donor used was Lot #20061357 from Hemacare. The gating strategy is mentioned in the figure. Lymphocytes derived from the donor were gated to remove dead cells. Live cells were further gated to remove doublets and other cell aggregates. Finally, CD8+ T cells were gated and the frequency of cells staining positive for both mutant and mimic peptide loaded tetramers (PE+/APC+) was determined. mi mu refers to a sample in which both mimic and mutant tetramers specific to the peptide were used for staining.

FIG. 7 illustrates exemplary FACS plots used to determine the frequency of dual positive T cells for tetramers specific for mutant and mimic peptides represented by SEQ ID NO: 3 and SEQ ID NO: 32. The donor used was Lot #20001476 from Hemacare. The gating strategy is mentioned in the figure. Lymphocytes derived from the donor were gated to remove dead cells. Live cells were further gated to remove doublets and other cell aggregates. Finally, CD8+ T cells were gated and the frequency of cells staining positive for both mutant and mimic peptide loaded tetramers (PE+/APC+) was determined. mi mu refers to a sample in which both mimic and mutant tetramers specific to the peptide were used for staining.

FIG. 8 illustrates exemplary FACS plots used to determine the frequency of dual positive T cells for tetramers specific for mutant and mimic peptides represented by SEQ ID NO: 16 and SEQ ID NO: 64. The donor used was Lot #20001476 from Hemacare. The gating strategy is mentioned in the figure. Lymphocytes derived from the donor were gated to remove dead cells. Live cells were further gated to remove doublets and other cell aggregates. Finally, CD8+ T cells were gated and the frequency of cells staining positive for both mutant and mimic peptide loaded tetramers (PE+/APC+) was determined. mi mu refers to a sample in which both mimic and mutant tetramers specific to the peptide were used for staining.

FIG. 9 illustrates exemplary FACS plots used to determine the frequency of dual positive T cells for tetramers specific for mutant and mimic peptides represented by SEQ ID NO: 23 and SEQ ID NO: 78. The donor used was Lot #20001476 from Hemacare. The gating strategy is mentioned in the figure. Lymphocytes derived from the donor were gated to remove dead cells. Live cells were further gated to remove doublets and other cell aggregates. Finally, CD8+ T cells were gated and the frequency of cells staining positive for both mutant and mimic peptide loaded tetramers (PE+/APC+) was determined. mi mu refers to a sample in which both mimic and mutant tetramers specific to the peptide were used for staining.

FIG. 10 illustrates exemplary FACS plots used to determine the frequency of dual positive T cells for tetramers specific for mutant and mimic peptides represented by SEQ ID NO: 22 and SEQ ID NO: 75. The donor used was Lot #20062224 from Hemacare. The gating strategy is mentioned in the figure. Lymphocytes derived from the donor were gated to remove dead cells. Live cells were further gated to remove doublets and other cell aggregates. Finally, CD8+ T cells were gated and the frequency of cells staining positive for both mutant and mimic peptide loaded tetramers (PE+/APC+) was determined. mi mu refers to a sample in which both mimic and mutant tetramers specific to the peptide were used for staining.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

The disclosed polypeptide fragments, polynucleotides, vectors, compositions, kits, methods, T-cell receptors (TCRs), and cells may be understood more readily by reference to the following detailed description taken in connection with the accompanying figures, which form a part of this disclosure. It is to be understood that the disclosed polypeptide fragments, polynucleotides, vectors, compositions, kits, methods, T-cell receptors (TCRs), and cells are not limited to those specifically described and/or shown herein, and that the terminology used herein is for the purpose of describing particular embodiments by way of example only and is not intended to be limiting of the claimed polypeptide fragments, polynucleotides, vectors, compositions, kits, methods, T-cell receptors (TCRs), and cells.

Unless specifically stated otherwise, any description as to a possible mechanism or mode of action or reason for improvement is meant to be illustrative only, and the disclosed polypeptide fragments, polynucleotides, vectors, compositions, kits, methods, T-cell receptors (TCRs), and cells are not to be constrained by the correctness or incorrectness of any such suggested mechanism or mode of action or reason for improvement.

Throughout this text, the descriptions refer to polypeptide fragments and methods of using said polypeptide fragments. Where the disclosure describes or claims a feature or embodiment associated with a polypeptide fragment, such a feature or embodiment is equally applicable to the methods of using said polypeptide fragment. Likewise, where the disclosure describes or claims a feature or embodiment associated with a method of using a polypeptide fragment, such a feature or embodiment is equally applicable to the polypeptide fragment.

Where a range of numerical values is recited or established herein, the range includes the endpoints thereof and all the individual integers and fractions within the range, and also includes each of the narrower ranges therein formed by all the various possible combinations of those endpoints and internal integers and fractions to form subgroups of the larger group of values within the stated range to the same extent as if each of those narrower ranges was explicitly recited. Where a range of numerical values is stated herein as being greater than a stated value, the range is nevertheless finite and is bounded on its upper end by a value that is operable within the context of the invention as described herein. Where a range of numerical values is stated herein as being less than a stated value, the range is nevertheless bounded on its lower end by a non-zero value. It is not intended that the scope of the invention be limited to the specific values recited when defining a range. All ranges are inclusive and combinable.

When values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. Reference to a particular numerical value includes at least that particular value, unless the context clearly dictates otherwise.

It is to be appreciated that certain features of the invention which are, for clarity, described herein in the context of separate embodiments may also be provided in combination in a single embodiment. That is, unless obviously incompatible or specifically excluded, each individual embodiment is deemed to be combinable with any other embodiment(s) and such a combination is considered to be another embodiment. Conversely, various features of the invention that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any sub-combination. Finally, although an embodiment may be described as part of a series of steps or part of a more general structure, each said step may also be considered an independent embodiment in itself, combinable with others.

Various terms relating to aspects of the description are used throughout the specification and claims. Such terms are to be given their ordinary meaning in the art unless otherwise indicated. Other specifically defined terms are to be construed in a manner consistent with the definitions provided herein.

The term “comprising” is intended to include examples encompassed by the terms “consisting essentially of” and “consisting of”; similarly, the term “consisting essentially of” is intended to include examples encompassed by the term “consisting of.”

When a value is expressed as an approximation by use of the descriptor “about,” it will be understood that the particular value forms another embodiment. In general, use of the term “about” indicates approximations that can vary depending on the desired properties sought to be obtained by the disclosed subject matter and is to be interpreted in the specific context in which it is used, based on its function. The person skilled in the art will be able to interpret this as a matter of routine. In some cases, the number of significant figures used for a particular value may be one non-limiting method of determining the extent of the word “about”. In other cases, the gradations used in a series of values may be used to determine the intended range available to the term “about” for each value. Where present, all ranges are inclusive and combinable. That is, references to values stated in ranges include every value within that range.

If not otherwise specified, the term “about” signifies a variance of ±10% of the associated value. Thus, the term “about” is used to encompass variations of ±10% or less, variations of ±5% or less, variations of ±1% or less, variations of ±0.5% or less, or variations of ±0.1% or less from the specified value.

When a list is presented, unless stated otherwise, it is to be understood that each individual element of that list, and every combination of that list, is a separate embodiment. For example, a list of embodiments presented as “A, B, or C” is to be interpreted as including the embodiments, “A”, “B”, “C”, “A or B”, “A or C”, “B or C”, or “A, B, or C”.

As used herein, the singular forms “a”, “an”, and “the” include the plural.

As used herein, the term “at least one” means “one or more.”

The terms “kit” and “article of manufacture” are used as synonyms.

“Neoantigen” refers to a mutated antigen which is expressed in tumor cells but not in normal cells. Neoantigens include antigens which arise from, for example, amino acid substitutions, frame shift mutation, fusion polypeptides, in-frame deletion, insertion, expression of endogenous retroviral polypeptides, and tumor-specific overexpression of polypeptides.

“9-mer” or “9mer” refers to a polypeptide that is nine amino acids in length.

“10-mer” or “10mer” refers to a polypeptide that is ten amino acids in length.

“Corresponding” refers to residues that occur at aligned loci. Related or variant polypeptides are aligned by any method known to those of skill in the art. Such methods typically maximize matches, and include methods such as using manual alignments and by using the numerous alignment programs available (for example, BLASTP) and others known to those of skill in the art. By aligning the sequences of polypeptides, one skilled in the art can identify corresponding residues, using conserved and identical amino acid residues as guides. Corresponding positions also can be based on structural alignments, for example by using computer simulated alignments of protein structure. In other instances, corresponding regions can be identified.

“Immunogenic fragment” refers to a polypeptide that is recognized by cytotoxic T lymphocytes, helper T lymphocytes or B cells when the fragment is in complex with MHC class I or MHC class II molecules.

“Subject” includes any human or nonhuman animal. “Nonhuman animal” includes all vertebrates, e.g., mammals and non-mammals, such as nonhuman primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, etc. The terms “subject” and “patient” can be used interchangeably herein.

“CIC” refers to human capicua transcriptional repressor. Human CIC protein comprises an amino acid sequence as shown for example in UniProt accession number Q96RK0.

“CTNNB1” refers to human catenin beta 1. Human CTTNB1 protein comprises an amino acid sequence as shown for example in UniProt accession number P35222.

“ERBB2” refers to human v-erb-b2 erythroblastic leukemia viral oncogene homolog B. Human ERBB2 protein comprises an amino acid sequence as shown for example in UniProt accession number P04626.

“KRAS” refers to human kirsten rat sarcoma. Human KRAS protein comprises an amino acid sequence as shown for example in UniProt accession number P01116.

“PIK3CA” refers to human phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha. Human PIK3CA protein comprises an amino acid sequence as shown for example in UniProt accession number P42336.

“PTEN” refers to human phosphatase and tensin homolog. Human PTEN comprises an amino acid sequence as shown for example in UniProt accession number P60484.

“SF3B1” refers to human splicing factor 3b subunit 1. Human SF3B1 comprises an amino acid sequence as shown for example in UniProt accession number 075533.

“SOX17” refers to human SRY-box transcription factor 17. Human SOX17 comprises an amino acid sequence as shown for example in UniProt accession number Q9H6I2.

“TP53” refers to human tumor protein 53. Human TP53 comprises an amino acid sequence as shown for example in UniProt accession number P04637.

“CMV” refers to human cytomegalovirus. Human CMV pp65 protein comprises an amino acid sequence as shown for example in UniProt accession number P18139.

Polypeptides

Provided herein are optimized MHC-binding polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to a cognate native MHC-binding polypeptide fragment. Exemplary modifications include, but are not limited to, substitutions, deletions or additions of amino acids. In certain embodiments, the MHC-binding polypeptides are neoantigens. In certain embodiments, the optimized immunogenic MHC-binding epitope has greater affinity for MHC than a cognate native MHC-binding polypeptide fragment. Peptide to MHC affinity (pMHC affinity) may range, for example, from >1 nM to <20,00 nM, with the strength of binding characterized as equilibrium dissociation constant, Kd (low dissociation constant represents high binding affinity)

Cognate native MHC-binding polypeptide fragments may be characterized by an absence of certain residues at critical anchor positions involved in MCH binding. Optimized MHC-binding polypeptide fragments may be generated by modifying amino acids at certain positions to improve MHC binding, for example as described in Slansky et al, Immunity, 2000 October; 13(4):529-38. In certain embodiments, the optimized MHC-binding polypeptide fragments is nine amino acids in length and comprises an amino acid substitution at amino acid position 2, amino acid position 9, or both. In certain embodiments, the optimized MHC-binding polypeptide fragments is ten amino acids in length and comprises an amino acid substitution at amino acid position 3, amino acid position 10, or both.

Positional numbering used herein (e.g. amino acid position 9) refers to the amino acid position starting at the N-terminus and moving toward the C-terminus. For purposes of illustration, taking the hypothetical peptide ABCDEFG, letter “A” is in amino acid position 1, letter “B” is in amino acid position 2, and so forth.

In some embodiments, the MHC-binding epitope binds to an MHC Class I or Class II molecule.

Preferred MHC Class I molecules include a heavy chain (e.g., an α chain) and a β2-microglobin. Such an MHC Class I molecule may be either a full-length molecule or an extracellular portion of a full-length molecule, such extracellular portion lacking complete transmembrane or cytoplasmic domains, or lacking both complete transmembrane and cytoplasmic domains. The MHC Class I molecule is preferably capable of binding a selected peptide. Exemplary MHC Class I molecules that may be employed in the present invention include, for example, molecules that are encoded by human leukocyte antigen (HLA)-A, HLA-B, HLA-C, HLA-E, HLA-F, or HLA-G loci. Preferably, the MHC Class I molecule is selected from molecules encoded by HLA-A, HLA-B, and HLA-C loci. Techniques, methods, and reagents useful for selection, cloning, preparation, and expression of β2-microglobin molecules, MHC Class I molecules such as HLA molecules, and portions thereof, are exemplified in U.S. Pat. Nos. 6,225,042, 6,355,479, and 6,362,001.

In certain embodiments, MHC Class I molecules include, but are not limited to, HLA 2.1 (HLA-A*02:01), HLA-A*01:01, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*33:03, HLA-C*07:01, HLA-C*07:02, HLA-C*04:01, HLA-B*07:02, HLA-B*44:02, and HLA-B*35:01. In preferred embodiments, MHC Class I molecule is HLA-A*02:01.

Preferred MHC Class II molecules include an alpha (α) chain and a beta (β) chain which associate together to form an MHC class II heterodimer. Such an MHC Class II heterodimer may be either a full-length molecule or an extracellular portion of a full-length α chain, an extracellular portion of a full-length β chain, or extracellular portions of both α and β chains, such extracellular portion or portions lacking complete transmembrane or cytoplasmic domains. Exemplary MHC Class II molecules that may be employed in the present invention include molecules that are encoded by HLA-DP, HLA-DQ HLA-DR, HLA-DO, HLA-DN, or HLA-DZ loci. Techniques, methods, and reagents useful for selection, cloning, preparation, and expression of MHC Class II α chains, β chains, and αβ heterodimers, and extracellular portions thereof, are exemplified in U.S. Pat. Nos. 5,583,031, and 6,355,479.

In certain embodiments, the neoantigen is encoded by a mutant variant of a gene selected from the group consisting of CIC, CTNNB1, ERBB2, KRAS, PIK3CA, PTEN, SF3B1, SOX17, TP53, and CMV.

In certain embodiments, the polypeptide fragment is selected from the group consisting of SEQ ID NO: 25, SEQ ID NO: 26 and SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, and SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81; SEQ ID NO: 84, SEQ ID NO: 85, and SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 91,SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, and any combination thereof.

In certain embodiments, the polypeptide fragment is selected from the group consisting of SEQ ID NO: 29, SEQ ID NO: 32, SEQ ID NO: 45, SEQ ID NO: 59, SEQ ID NO: 64, SEQ ID NO: 68, SEQ ID NO: 75, SEQ ID NO: 78, and any combination thereof.

CIC Polypeptides

Described herein are CIC polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 102, and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length. Described herein are CIC polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 102, and an amino acid substitution at amino acid position 3 of the fragment, amino acid position 10 of the fragment, or both, wherein the fragment is at least ten amino acids in length.

In certain embodiments, the modification comprises a deletion, insertion, and/or substitution. In preferred embodiments, the modification comprises a substitution. In more preferred embodiments, the modification comprises an arginine to tryptophan amino acid substitution at a position corresponding to position 215 of SEQ ID NO: 102 (R215W).

Described herein are CIC polypeptide fragments comprising, consisting of, or consisting essentially of an arginine to tryptophan amino acid substitution at a position corresponding to position 215 of SEQ ID NO: 102 (R215W), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the CIC polypeptide fragments comprise, consist of, or consist essentially of an arginine to tryptophan amino acid substitution at a position corresponding to position 215 of SEQ ID NO: 102 (R215W), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the CIC polypeptide fragments comprise, consist of, or consist essentially of an arginine to tryptophan amino acid substitution at a position corresponding to position 215 of SEQ ID NO: 102 (R215W), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the CIC polypeptide fragments comprise, consist of, or consist essentially of an arginine to tryptophan amino acid substitution at a position corresponding to position 215 of SEQ ID NO: 102 (R215W), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

In certain embodiments, the CIC polypeptide fragment binds to HLA 2.1 (HLA-A*02:01), HLA-A*01:01, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*33:03, HLA-C*07:01, HLA-C*07:02, HLA-C*04:01, HLA-B*07:02, HLA-B*44:02, or HLA-B*35:01. In preferred embodiments, the CIC polypeptide fragment binds to HLA-A*02:01.

In some embodiments, the CIC polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control, expressed as a percent (%) binding affinity. In certain embodiments, the CIC polypeptide fragment has a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment, wherein binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control.

In certain embodiments, the R215W substitution is at amino acid position 1 of the fragment. In certain embodiments, the R215W substitution is at amino acid position 2 of the fragment. In certain embodiments, the R215W substitution is at amino acid position 3 of the fragment. In certain embodiments, the R215W substitution is at amino acid position 4 of the fragment. In certain embodiments, the R215W substitution is at amino acid position 5 of the fragment. In certain embodiments, the R215W substitution is at amino acid position 6 of the fragment. In certain embodiments, the R215W substitution is at amino acid position 7 of the fragment. In certain embodiments, the R215W substitution is at amino acid position 8 of the fragment. In certain embodiments, the R215W substitution is at amino acid position 9 of the fragment. In certain embodiments, the R215W substitution is at amino acid position 10 of the fragment. In preferred embodiments, the R215W substitution is at amino acid position 8 of the fragment.

In further embodiments, the CIC polypeptide fragment is selected from the group consisting of SEQ ID NO: 25, SEQ ID NO: 26 and SEQ ID NO: 27. In certain embodiments, the CIC polypeptide fragment is SEQ ID NO: 25 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 25. In certain embodiments, the CIC polypeptide fragment is SEQ ID NO: 26 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 26. In certain embodiments, the CIC polypeptide fragment is SEQ ID NO: 27 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 27.

In some embodiments, the CIC polypeptide fragment has the sequence MX₂FSKRHWX₉ (SEQ ID NO: 225), wherein X₂ is any amino acid other than isoleucine, and preferably methionine or leucine, and X₉ is any amino acid other than alanine, and preferably isoleucine or valine.

CTNNB1 Polypeptides

Described herein are CTNNB1 polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 103, and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length. Described herein are CTNNB1 polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 103, and an amino acid substitution at amino acid position 3 of the fragment, amino acid position 10 of the fragment, or both, wherein the fragment is at least ten amino acids in length.

In certain embodiments, the modification comprises a deletion, insertion, and/or substitution. In preferred embodiments, the modification comprises a substitution. In further preferred embodiments, the modification comprises a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C). In further preferred embodiments, the modification comprises a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F).

Described herein are CTNNB1 polypeptide fragments comprising, consisting of, or consisting essentially of a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the CTNNB1 polypeptide fragments comprise, consist of, or consist essentially of a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the CTNNB1 polypeptide fragments comprise, consist of, or consist essentially of a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the CTNNB1 polypeptide fragments comprise, consist of, or consist essentially of a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

Described herein are CTNNB1 polypeptide fragments comprising, consisting of, or consisting essentially of a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the CTNNB1 polypeptide fragments comprise, consist of, or consist essentially of a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the CTNNB1 polypeptide fragments comprise, consist of, or consist essentially of a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the CTNNB1 polypeptide fragments comprise, consist of, or consist essentially of a serine to phenylalanine acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

In certain embodiments, the CTNNB1 polypeptide fragment binds to HLA 2.1 (HLA-A*02:01), HLA-A*01:01, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*33:03, HLA-C*07:01, HLA-C*07:02, HLA-C*04:01, HLA-B*07:02, HLA-B*44:02, or HLA-B*35:01. In preferred embodiments, the CTNNB1 polypeptide fragment binds to HLA-A*02:01.

In some embodiments, the CTNNB1 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control, expressed as a percent (%) binding affinity. In certain embodiments, the CTNNB1 polypeptide fragment has a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment, wherein binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control.

In certain embodiments, the S33C substitution is at amino acid position 1 of the fragment. In certain embodiments, the S33C substitution is at amino acid position 2 of the fragment. In certain embodiments, the S33C substitution is at amino acid position 3 of the fragment. In certain embodiments, the S33C substitution is at amino acid position 4 of the fragment. In certain embodiments, the S33C substitution is at amino acid position 5 of the fragment. In certain embodiments, the S33C substitution is at amino acid position 6 of the fragment. In certain embodiments, the S33C substitution is at amino acid position 7 of the fragment. In certain embodiments, the S33C substitution is at amino acid position 8 of the fragment. In certain embodiments, the S33C substitution is at amino acid position 9 of the fragment. In certain embodiments, the S33C substitution is at amino acid position 10 of the fragment. In preferred embodiments, the S33C substitution is at amino acid position 4 of the fragment.

In certain embodiments, the S37F substitution is at amino acid position 1 of the fragment. In certain embodiments, the S37F substitution is at amino acid position 2 of the fragment. In certain embodiments, the S37F substitution is at amino acid position 3 of the fragment. In certain embodiments, the S37F substitution is at amino acid position 4 of the fragment. In certain embodiments, the S37F substitution is at amino acid position 5 of the fragment. In certain embodiments, the S37F substitution is at amino acid position 6 of the fragment. In certain embodiments, the S37F substitution is at amino acid position 7 of the fragment. In certain embodiments, the S37F substitution is at amino acid position 8 of the fragment. In certain embodiments, the S37F substitution is at amino acid position 9 of the fragment. In certain embodiments, the S37F substitution is at amino acid position 10 of the fragment. In preferred embodiments, the S37F substitution is at amino acid position 8 of the fragment.

In still further embodiments, the CTNNB1 polypeptide fragment is selected from the group consisting of SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 80, and SEQ ID NO: 81. In still further embodiments, the CTNNB1 polypeptide fragment is SEQ ID NO: 28 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 28. In still further embodiments, the CTNNB1 polypeptide fragment is SEQ ID NO: 29 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 29. In still further embodiments, the CTNNB1 polypeptide fragment is SEQ ID NO: 30 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 30. In still further embodiments, the CTNNB1 polypeptide fragment is SEQ ID NO: 31 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 31. In still further embodiments, the CTNNB1 polypeptide fragment is SEQ ID NO: 32 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 32. In still further embodiments, the CTNNB1 polypeptide fragment is SEQ ID NO: 33 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 33. In still further embodiments, the CTNNB1 polypeptide fragment is SEQ ID NO: 80 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 80. In still further embodiments, the CTNNB1 polypeptide fragment is SEQ ID NO: 81 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 81.

In some embodiments, the CTNNB1 polypeptide fragment has the sequence YX₂DCGIHSX₉ (SEQ ID NO: 226), wherein X₂ is any amino acid other than leucine, and preferably methionine, and X₉ is any amino acid other than glycine, and preferably leucine or valine.

In some embodiments, the CTNNB1 polypeptide fragment has the sequence YX₂DSGIHFX₉ (SEQ ID NO: 227), wherein X₂ is any amino acid other than leucine, and preferably methionine, and X₉ is any amino acid other than glycine, and preferably isoleucine, leucine or valine.

KRAS Polypeptides

Described herein are KRAS polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 105, and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length. Described herein are KRAS polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 105, and an amino acid substitution at amino acid position 3 of the fragment, amino acid position 10 of the fragment, or both, wherein the fragment is at least ten amino acids in length.

In certain embodiments, the modification comprises a deletion, insertion, and/or substitution. In preferred embodiments, the modification comprises a substitution. In further preferred embodiments, the modification comprises a glycine to alanine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12A). In further preferred embodiments, the modification comprises a glycine to cysteine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12C). In further preferred embodiments, the modification comprises a glycine to valine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12V).

Described herein are KRAS polypeptide fragments comprising, consisting of, or consisting essentially of a glycine to alanine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12A), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the KRAS polypeptide fragments comprise, consist of, or consist essentially of a glycine to alanine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12A), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the KRAS polypeptide fragments comprise, consist of, or consist essentially of a glycine to alanine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12A), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the KRAS polypeptide fragments comprise, consist of, or consist essentially of a glycine to alanine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12A), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

Described herein are KRAS polypeptide fragments comprising, consisting of, or consisting essentially of a glycine to cysteine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12C), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the KRAS polypeptide fragments comprise, consist of, or consist essentially of a glycine to cysteine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12C), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the KRAS polypeptide fragments comprise, consist of, or consist essentially of a glycine to cysteine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12C), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the KRAS polypeptide fragments comprise, consist of, or consist essentially of a glycine to cysteine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12C), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

Described herein are KRAS polypeptide fragments comprising, consisting of, or consisting essentially of a glycine to valine amino acid substitution at a position corresponding to at position 12 of SEQ ID NO: 105 (G12V), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the KRAS polypeptide fragments comprise, consist of, or consist essentially of a glycine to valine amino acid substitution at a position corresponding to at position 12 of SEQ ID NO: 105 (G12V), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the KRAS polypeptide fragments comprise, consist of, or consist essentially of a glycine to valine amino acid substitution at a position corresponding to at position 12 of SEQ ID NO: 105 (G12V), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the KRAS polypeptide fragments comprise, consist of, or consist essentially of a glycine to valine amino acid substitution at a position corresponding to at position 12 of SEQ ID NO: 105 (G12V), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

In certain embodiments, the KRAS polypeptide fragment binds to HLA 2.1 (HLA-A*02:01), HLA-A*01:01, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*33:03, HLA-C*07:01, HLA-C*07:02, HLA-C*04:01, HLA-B*07:02, HLA-B*44:02, or HLA-B*35:01. In preferred embodiments, the KRAS polypeptide fragment binds to HLA-A*02:01.

In some embodiments, the KRAS polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control, expressed as a percent (%) binding affinity. In certain embodiments, the KRAS polypeptide fragment has a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment, wherein binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control.

In certain embodiments, the G12A substitution is at amino acid position 1 of the fragment. In certain embodiments, the G12A substitution is at amino acid position 2 of the fragment. In certain embodiments, the G12A substitution is at amino acid position 3 of the fragment. In certain embodiments, the G12A substitution is at amino acid position 4 of the fragment. In certain embodiments, the G12A substitution is at amino acid position 5 of the fragment. In certain embodiments, the G12A substitution is at amino acid position 6 of the fragment. In certain embodiments, the G12A substitution is at amino acid position 7 of the fragment. In certain embodiments, the G12A substitution is at amino acid position 8 of the fragment. In certain embodiments, the G12A substitution is at amino acid position 9 of the fragment. In certain embodiments, the G12A substitution is at amino acid position 10 of the fragment. In preferred embodiments, the G12A substitution is at amino acid position 7 of the fragment.

In certain embodiments, the G12C substitution is at amino acid position 1 of the fragment. In certain embodiments, the G12C substitution is at amino acid position 2 of the fragment. In certain embodiments, the G12C substitution is at amino acid position 3 of the fragment. In certain embodiments, the G12C substitution is at amino acid position 4 of the fragment. In certain embodiments, the G12C substitution is at amino acid position 5 of the fragment. In certain embodiments, the G12C substitution is at amino acid position 6 of the fragment. In certain embodiments, the G12C substitution is at amino acid position 7 of the fragment. In certain embodiments, the G12C substitution is at amino acid position 8 of the fragment. In certain embodiments, the G12C substitution is at amino acid position 9 of the fragment. In certain embodiments, the G12C substitution is at amino acid position 10 of the fragment. In preferred embodiments, the G12C substitution is at amino acid position 7 of the fragment.

In certain embodiments, the G12V substitution is at amino acid position 1 of the fragment. In certain embodiments, the G12V substitution is at amino acid position 2 of the fragment. In certain embodiments, the G12V substitution is at amino acid position 3 of the fragment. In certain embodiments, the G12V substitution is at amino acid position 4 of the fragment. In certain embodiments, the G12V substitution is at amino acid position 5 of the fragment. In certain embodiments, the G12V substitution is at amino acid position 6 of the fragment. In certain embodiments, the G12V substitution is at amino acid position 7 of the fragment. In certain embodiments, the G12V substitution is at amino acid position 8 of the fragment. In certain embodiments, the G12V substitution is at amino acid position 9 of the fragment. In certain embodiments, the G12V substitution is at amino acid position 10 of the fragment. In preferred embodiments, the G12V substitution is at amino acid position 7 of the fragment.

In some embodiments, the KRAS polypeptide fragment is selected from the group consisting of SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, and SEQ ID NO: 42. In certain embodiments, the KRAS polypeptide fragment is SEQ ID NO: 37 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 37. In certain embodiments, the KRAS polypeptide fragment is SEQ ID NO: 38 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 38. In certain embodiments, the KRAS polypeptide fragment is SEQ ID NO: 39 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 39. In certain embodiments, the KRAS polypeptide fragment is SEQ ID NO: 40 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 40. In certain embodiments, the KRAS polypeptide fragment is SEQ ID NO: 41 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 41. In certain embodiments, the KRAS polypeptide fragment is SEQ ID NO: 42 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 42.

In some embodiments, the KRAS polypeptide fragment has the sequence LX₂VVGAAGV (SEQ ID NO: 228), wherein X₂ is any amino acid other than valine, and preferably methionine or leucine.

In some embodiments, the KRAS polypeptide fragment has the sequence LX₂VVGACGV (SEQ ID NO: 229), wherein X₂ is any amino acid other than valine, and preferably methionine or leucine.

In some embodiments, the KRAS polypeptide fragment has the sequence LX₂VVGAVGV (SEQ ID NO: 230), wherein X₂ is any amino acid other than valine, and preferably methionine or leucine.

PIK3CA Polypeptides

Described herein are PIK3CA polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 106, and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length. Described herein are PIK3CA polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 106, and an amino acid substitution at amino acid position 3 of the fragment, amino acid position 10 of the fragment, or both, wherein the fragment is at least ten amino acids in length.

In certain embodiments, the modification comprises a deletion, insertion, and/or substitution. In preferred embodiments, the modification comprises a substitution. In further preferred embodiments, the modification comprises a glutamic acid to lysine amino acid substitution at a position corresponding to position 453 of SEQ ID NO: 106 (E453K). In further preferred embodiments, the modification comprises a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D).

Described herein are PIK3CA polypeptide fragments comprising, consisting of, or consisting essentially of a glutamic acid to lysine amino acid substitution at a position corresponding to position 453 of SEQ ID NO: 106 (E453K), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the PIK3CA polypeptide fragments comprise, consist of, or consist essentially of a glutamic acid to lysine amino acid substitution at a position corresponding to position 453 of SEQ ID NO: 106 (E453K), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the PIK3CA polypeptide fragments comprise, consist of, or consist essentially of a glutamic acid to lysine amino acid substitution at a position corresponding to position 453 of SEQ ID NO: 106 (E453K), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the PIK3CA polypeptide fragments comprise, consist of, or consist essentially of a glutamic acid to lysine amino acid substitution at a position corresponding to position 453 of SEQ ID NO: 106 (E453K), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

Described herein are PIK3CA polypeptide fragments comprising, consisting of, or consisting essentially of a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the PIK3CA polypeptide fragments comprise, consist of, or consist essentially of a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the PIK3CA polypeptide fragments comprise, consist of, or consist essentially of a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the PIK3CA polypeptide fragments comprise, consist of, or consist essentially of a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

In certain embodiments, the PIK3CA polypeptide fragment binds to HLA 2.1 (HLA-A*02:01), HLA-A*01:01, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*33:03, HLA-C*07:01, HLA-C*07:02, HLA-C*04:01, HLA-B*07:02, HLA-B*44:02, or HLA-B*35:01. In preferred embodiments, the PIK3CA polypeptide fragment binds to HLA-A*02:01.

In some embodiments, the PIK3CA polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control, expressed as a percent (%) binding affinity. In certain embodiments, the PIK3CA polypeptide fragment has a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment, wherein binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control.

In certain embodiments, the E453K substitution is at amino acid position 1 of the fragment. In certain embodiments, the E453K substitution is at amino acid position 2 of the fragment. In certain embodiments, the E453K substitution is at amino acid position 3 of the fragment. In certain embodiments, the E453K substitution is at amino acid position 4 of the fragment. In certain embodiments, the E453K substitution is at amino acid position 5 of the fragment. In certain embodiments, the E453K substitution is at amino acid position 6 of the fragment. In certain embodiments, the E453K substitution is at amino acid position 7 of the fragment. In certain embodiments, the E453K substitution is at amino acid position 8 of the fragment. In certain embodiments, the E453K substitution is at amino acid position 9 of the fragment. In certain embodiments, the E453K substitution is at amino acid position 10 of the fragment. In preferred embodiments, the E453K substitution is at amino acid position 3 of the fragment.

In certain embodiments, the G118D substitution is at amino acid position 1 of the fragment. In certain embodiments, the G118D substitution is at amino acid position 2 of the fragment. In certain embodiments, the G118D substitution is at amino acid position 3 of the fragment. In certain embodiments, the G118D substitution is at amino acid position 4 of the fragment. In certain embodiments, the G118D substitution is at amino acid position 5 of the fragment. In certain embodiments, the G118D substitution is at amino acid position 6 of the fragment. In certain embodiments, the G118D substitution is at amino acid position 7 of the fragment. In certain embodiments, the G118D substitution is at amino acid position 8 of the fragment. In certain embodiments, the G118D substitution is at amino acid position 9 of the fragment. In certain embodiments, the G118D substitution is at amino acid position 10 of the fragment. In preferred embodiments, the G118D substitution is at amino acid position 7 of the fragment.

In still further embodiments, the PIK3CA polypeptide fragment is selected from the group consisting of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, and SEQ ID NO: 47. In certain embodiments, the PIK3CA polypeptide fragment is SEQ ID NO: 43 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 43. In certain embodiments, the PIK3CA polypeptide fragment is SEQ ID NO: 44 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 44. In certain embodiments, the PIK3CA polypeptide fragment is SEQ ID NO: 45 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 45. In certain embodiments, the PIK3CA polypeptide fragment is SEQ ID NO: 46 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 46. In certain embodiments, the PIK3CA polypeptide fragment is SEQ ID NO: 47 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 47.

In some embodiments, the PIK3CA polypeptide fragment has the sequence GX₂KDLLNPX₉ (SEQ ID NO: 231), wherein X₂ is any amino acid other than leucine, and preferably methionine, and X9 is any amino acid other than isoleucine, and preferably valine.

In some embodiments, the PIK3CA polypeptide fragment has the sequence IX₂NREIDFX₉ (SEQ ID NO: 232), wherein X₂ is any amino acid other than leucine, and preferably methionine, and X₉ is any amino acid other than alanine, and preferably valine or leucine.

PTEN Polypeptides

Described herein are PTEN polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 107, and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length. Described herein are PTEN polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 107, and an amino acid substitution at amino acid position 3 of the fragment, amino acid position 10 of the fragment, or both, wherein the fragment is at least ten amino acids in length.

In certain embodiments, the modification comprises a deletion, insertion, and/or substitution. In preferred embodiments, the modification comprises a substitution. In further preferred embodiments, the modification comprises an arginine to cysteine amino acid substitution at a position corresponding to position 173 of SEQ ID NO: 107 (R173C).

Described herein are PTEN polypeptide fragments comprising, consisting of, or consisting essentially of an arginine to cysteine amino acid substitution at a position corresponding to position 173 of SEQ ID NO: 107 (R173C), and an amino acid substitution at amino acid position 3 of the fragment, amino acid position 10 of the fragment, or both, wherein the fragment is ten amino acids in length. In certain embodiments, the PTEN polypeptide fragments comprise, consist of, or consist essentially of an arginine to cysteine amino acid substitution at a position corresponding to position 173 of SEQ ID NO: 107 (R173C), and an amino acid substitution at amino acid position 3 of the fragment. In certain embodiments, the PTEN polypeptide fragments comprise, consist of, or consist essentially of an arginine to cysteine amino acid substitution at a position corresponding to position 173 of SEQ ID NO: 107 (R173C), and an amino acid substitution at amino acid position 10 of the fragment. In certain embodiments, the PTEN polypeptide fragments comprise, consist of, or consist essentially of an arginine to cysteine amino acid substitution at a position corresponding to position 173 of SEQ ID NO: 107 (R173C), and an amino acid substitution at amino acid position 3 of the fragment and at position 10 of the fragment.

In certain embodiments, the PTEN polypeptide fragment binds to HLA 2.1 (HLA-A*02:01), HLA-A*01:01, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*33:03, HLA-C*07:01, HLA-C*07:02, HLA-C*04:01, HLA-B*07:02, HLA-B*44:02, or HLA-B*35:01. In preferred embodiments, the PTEN polypeptide fragment binds to HLA-A*02:01.

In some embodiments, the PTEN polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control, expressed as a percent (%) binding affinity. In certain embodiments, the PTEN polypeptide fragment has a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment, wherein binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control.

In certain embodiments, the R173C substitution is at amino acid position 1 of the fragment. In certain embodiments, the R173C substitution is at amino acid position 2 of the fragment. In certain embodiments, the R173C substitution is at amino acid position 3 of the fragment. In certain embodiments, the R173C substitution is at amino acid position 4 of the fragment. In certain embodiments, the R173C substitution is at amino acid position 5 of the fragment. In certain embodiments, the R173C substitution is at amino acid position 6 of the fragment. In certain embodiments, the R173C substitution is at amino acid position 7 of the fragment. In certain embodiments, the R173C substitution is at amino acid position 8 of the fragment. In certain embodiments, the R173C substitution is at amino acid position 9 of the fragment. In certain embodiments, the R173C substitution is at amino acid position 10 of the fragment. In preferred embodiments, the R173C substitution is at amino acid position 1 of the fragment.

In further embodiments, the PTEN polypeptide fragment is selected from the group consisting of SEQ ID NO: 48, SEQ ID NO: 49, and SEQ ID NO: 88. In certain embodiments, the PTEN polypeptide fragment is SEQ ID NO: 48 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 48. In certain embodiments, the PTEN polypeptide fragment is SEQ ID NO: 49 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 49. In certain embodiments, the PTEN polypeptide fragment is SEQ ID NO: 88 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 88.

In some embodiments, the PTEN polypeptide fragment has the sequence CYX₃YYYSYLX₁₀ (SEQ ID NO: 233), wherein X₃ is any amino acid other than valine, and preferably methionine or leucine, and X₁₀ is any amino acid other than leucine, and preferably valine or isoleucine.

SF3B1 Polypeptides

Described herein are SF3B1 polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 108, and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length. Described herein are SF3B1 polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 108, and an amino acid substitution at amino acid position 3 of the fragment, amino acid position 10 of the fragment, or both, wherein the fragment is at least ten amino acids in length.

In certain embodiments, the modification comprises a deletion, insertion, and/or substitution. In preferred embodiments, the modification comprises a substitution. In further preferred embodiments, the modification comprises an arginine to histidine amino acid substitution at a position corresponding to position 625 of SEQ ID NO: 108 (R625H).

Described herein are SF3B1 polypeptide fragments comprising, consisting of, or consisting essentially of an arginine to histidine amino acid substitution at a position corresponding to position 625 of SEQ ID NO: 108 (R625H), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the SF3B1 polypeptide fragments comprise, consist of, or consist essentially of an arginine to histidine amino acid substitution at a position corresponding to position 625 of SEQ ID NO: 108 (R625H), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the SF3B1 polypeptide fragments comprise, consist of, or consist essentially of an arginine to histidine amino acid substitution at a position corresponding to position 625 of SEQ ID NO: 108 (R625H), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the SF3B1 polypeptide fragments comprise, consist of, or consist essentially of an arginine to histidine amino acid substitution at a position corresponding to position 625 of SEQ ID NO: 108 (R625H), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

In certain embodiments, the SF3B1 polypeptide fragment binds to HLA 2.1 (HLA-A*02:01), HLA-A*01:01, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*33:03, HLA-C*07:01, HLA-C*07:02, HLA-C*04:01, HLA-B*07:02, HLA-B*44:02, or HLA-B*35:01. In preferred embodiments, the SF3B1 polypeptide fragment binds to HLA-A*02:01.

In some embodiments, the SF3B1 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control, expressed as a percent (%) binding affinity. In certain embodiments, the SF3B1 polypeptide fragment has a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment, wherein binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control.

In certain embodiments, the R625H substitution is at amino acid position 1 of the fragment. In certain embodiments, the R625H substitution is at amino acid position 2 of the fragment. In certain embodiments, the R625H substitution is at amino acid position 3 of the fragment. In certain embodiments, the R625H substitution is at amino acid position 4 of the fragment. In certain embodiments, the R625H substitution is at amino acid position 5 of the fragment. In certain embodiments, the R625H substitution is at amino acid position 6 of the fragment. In certain embodiments, the R625H substitution is at amino acid position 7 of the fragment. In certain embodiments, the R625H substitution is at amino acid position 8 of the fragment. In certain embodiments, the R625H substitution is at amino acid position 9 of the fragment. In certain embodiments, the R625H substitution is at amino acid position 10 of the fragment. In preferred embodiments, the R625H substitution is at amino acid position 7 of the fragment.

In further embodiments, the SF3B1 polypeptide fragment is selected from the group consisting of SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 90, SEQ ID NO: 91 and SEQ ID NO: 92. In certain embodiments, the SF3B1 polypeptide fragment is SEQ ID NO: 51 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 51. In certain embodiments, the SF3B1 polypeptide fragment is SEQ ID NO: 52 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 52. In certain embodiments, the SF3B1 polypeptide fragment is SEQ ID NO: 53 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 53. In certain embodiments, the SF3B1 polypeptide fragment is SEQ ID NO: 90 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 90. In certain embodiments, the SF3B1 polypeptide fragment is SEQ ID NO: 91 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 91. In certain embodiments, the SF3B1 polypeptide fragment is SEQ ID NO: 92 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 92.

In some embodiments, the SF3B1 polypeptide fragment has the sequence NX₂DEYVHNX₉ (SEQ ID NO: 234), wherein X₂ is any amino acid other than methionine, and preferably leucine, and X₉ is any amino acid other than threonine, and preferably valine, leucine, or isoleucine.

SOX17 Polypeptides

Described herein are SOX17 polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 109, and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length. Described herein are SOX17 polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 109, and an amino acid substitution at amino acid position 3 of the fragment, amino acid position 10 of the fragment, or both, wherein the fragment is at least ten amino acids in length.

In certain embodiments, the modification comprises a deletion, insertion, and/or substitution. In preferred embodiments, the modification comprises a substitution. In further preferred embodiments, the modification comprises a serine to isoleucine amino acid substitution at a position corresponding to position 403 of SEQ ID NO: 109 (S403I).

Described herein are SOX17 polypeptide fragments comprising, consisting of, or consisting essentially of a serine to isoleucine amino acid substitution at a position corresponding to position 403 of SEQ ID NO: 109 (S403I), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the SOX17 polypeptide fragments comprise, consist of, or consist essentially of a serine to isoleucine amino acid substitution at a position corresponding to position 403 of SEQ ID NO: 109 (S403I), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the SOX17 polypeptide fragments comprise, consist of, or consist essentially of a serine to isoleucine amino acid substitution at a position corresponding to position 403 of SEQ ID NO: 109 (S403I), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the SOX17 polypeptide fragments comprise, consist of, or consist essentially of a serine to isoleucine amino acid substitution at a position corresponding to position 403 of SEQ ID NO: 109 (S403I), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

In certain embodiments, the SOX17 polypeptide fragment binds to HLA 2.1 (HLA-A*02:01), HLA-A*01:01, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*33:03, HLA-C*07:01, HLA-C*07:02, HLA-C*04:01, HLA-B*07:02, HLA-B*44:02, or HLA-B*35:01. In preferred embodiments, the SOX17 polypeptide fragment binds to HLA-A*02:01.

In some embodiments, the SOX17 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control, expressed as a percent (%) binding affinity. In certain embodiments, the SOX17 polypeptide fragment has a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment, wherein binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control.

In certain embodiments, the S403I substitution is at amino acid position 1 of the fragment. In certain embodiments, the S403I substitution is at amino acid position 2 of the fragment. In certain embodiments, the S403I substitution is at amino acid position 3 of the fragment. In certain embodiments, the S403I substitution is at amino acid position 4 of the fragment. In certain embodiments, the S403I substitution is at amino acid position 5 of the fragment. In certain embodiments, the S403I substitution is at amino acid position 6 of the fragment. In certain embodiments, the S403I substitution is at amino acid position 7 of the fragment. In certain embodiments, the S403I substitution is at amino acid position 8 of the fragment. In certain embodiments, the S403I substitution is at amino acid position 9 of the fragment. In certain embodiments, the S403I substitution is at amino acid position 10 of the fragment. In preferred embodiments, the S403I substitution is at amino acid position 6 of the fragment.

In further embodiments, the SOX17 polypeptide fragment is selected from the group consisting of SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, and SEQ ID NO: 93. In certain embodiments, the SOX17 polypeptide fragment is SEQ ID NO: 54 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 54. In certain embodiments, the SOX17 polypeptide fragment is SEQ ID NO: 55 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 55. In certain embodiments, the SOX17 polypeptide fragment is SEQ ID NO: 56 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 56. In certain embodiments, the SOX17 polypeptide fragment is SEQ ID NO: 93 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 93.

In some embodiments, the SOX17 polypeptide fragment has the sequence VX₂SDAISAX₉ (SEQ ID NO: 235), wherein X₂ is any amino acid other than valine, and preferably leucine or methionine, and X₉ is any amino acid other than valine, and preferably leucine.

TP53 Polypeptides

Described herein are TP53 polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 110, and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length. Described herein are TP53 polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 110, and an amino acid substitution at amino acid position 3 of the fragment, amino acid position 10 of the fragment, or both, wherein the fragment is at least ten amino acids in length.

In certain embodiments, the modification comprises a deletion, insertion, and/or substitution. In preferred embodiments, the modification comprises a substitution. In further preferred embodiments, the modification comprises an arginine to leucine amino acid substitution at a position corresponding to position 110 of SEQ ID NO: 110 (R110L). In further preferred embodiments, the modification comprises a serine to phenylalanine amino acid substitution at a position corresponding to position 127 of SEQ ID NO: 110 (S127F). In further preferred embodiments, the modification comprises a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N). In further preferred embodiments, the modification comprises a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y). In further preferred embodiments, the modification comprises a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L). In further preferred embodiments, the modification comprises a histidine to leucine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193L). In further preferred embodiments, the modification comprises a histidine to tyrosine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193Y). In further preferred embodiments, the modification comprises a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C). In further preferred embodiments, the modification comprises a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M).

Described herein are TP53 polypeptide fragments comprising, consisting of, or consisting essentially of an arginine to leucine amino acid substitution at a position corresponding to position 110 of SEQ ID NO: 110 (R110L), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of an arginine to leucine amino acid substitution at a position corresponding to position 110 of SEQ ID NO: 110 (R110L), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of an arginine to leucine amino acid substitution at a position corresponding to position 110 of SEQ ID NO: 110 (R110L), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of an arginine to leucine amino acid substitution at a position corresponding to position 110 of SEQ ID NO: 110 (R110L), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

Described herein are TP53 polypeptide fragments comprising, consisting of, or consisting essentially of a serine to phenylalanine amino acid substitution at a position corresponding to position 127 of SEQ ID NO: 110 (S127F), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a serine to phenylalanine amino acid substitution at a position corresponding to position 127 of SEQ ID NO: 110 (S127F), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a serine to phenylalanine amino acid substitution at a position corresponding to position 127 of SEQ ID NO: 110 (S127F), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a serine to phenylalanine amino acid substitution at a position corresponding to position 127 of SEQ ID NO: 110 (S127F), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

Described herein are TP53 polypeptide fragments comprising, consisting of, or consisting essentially of a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

Described herein are TP53 polypeptide fragments comprising, consisting of, or consisting essentially of a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

Described herein are TP53 polypeptide fragments comprising, consisting of, or consisting essentially of a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

Described herein are TP53 polypeptide fragments comprising, consisting of, or consisting essentially of a histidine to leucine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193L), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a histidine to leucine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193L), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a histidine to leucine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193L), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a histidine to leucine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193L), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

Described herein are TP53 polypeptide fragments comprising, consisting of, or consisting essentially of a histidine to tyrosine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193Y), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a histidine to tyrosine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193Y), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a histidine to tyrosine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193Y), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a histidine to tyrosine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193Y), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

Described herein are TP53 polypeptide fragments comprising, consisting of, or consisting essentially of a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

Described herein are TP53 polypeptide fragments comprising, consisting of, or consisting essentially of a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the TP53 polypeptide fragments comprise, consist of, or consist essentially of a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

In certain embodiments, the TP53 polypeptide fragment binds to HLA 2.1 (HLA-A*02:01), HLA-A*01:01, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*33:03, HLA-C*07:01, HLA-C*07:02, HLA-C*04:01, HLA-B*07:02, HLA-B*44:02, or HLA-B*35:01. In preferred embodiments, the SOX17 polypeptide fragment binds to HLA-A*02:01.

In some embodiments, the TP53 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control, expressed as a percent (%) binding affinity. In certain embodiments, the TP53 polypeptide fragment has a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment, wherein binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control.

In certain embodiments, the R110L substitution is at amino acid position 1 of the fragment. In certain embodiments, the R110L substitution is at amino acid position 2 of the fragment. In certain embodiments, the R110L substitution is at amino acid position 3 of the fragment. In certain embodiments, the R110L substitution is at amino acid position 4 of the fragment. In certain embodiments, the R110L substitution is at amino acid position 5 of the fragment. In certain embodiments, the R110L substitution is at amino acid position 6 of the fragment. In certain embodiments, the R110L substitution is at amino acid position 7 of the fragment. In certain embodiments, the R110L substitution is at amino acid position 8 of the fragment. In certain embodiments, the R110L substitution is at amino acid position 9 of the fragment. In certain embodiments, the R110L substitution is at amino acid position 10 of the fragment. In preferred embodiments, the R110L substitution is at amino acid position 8 of the fragment.

In certain embodiments, the S127F substitution is at amino acid position 1 of the fragment. In certain embodiments, the S127F substitution is at amino acid position 2 of the fragment. In certain embodiments, the S127F substitution is at amino acid position 3 of the fragment. In certain embodiments, the S127F substitution is at amino acid position 4 of the fragment. In certain embodiments, the S127F substitution is at amino acid position 5 of the fragment. In certain embodiments, the S127F substitution is at amino acid position 6 of the fragment. In certain embodiments, the S127F substitution is at amino acid position 7 of the fragment. In certain embodiments, the S127F substitution is at amino acid position 8 of the fragment. In certain embodiments, the S127F substitution is at amino acid position 9 of the fragment. In certain embodiments, the S127F substitution is at amino acid position 10 of the fragment. In preferred embodiments, the S127F substitution is at amino acid position 7 of the fragment.

In certain embodiments, the K132N substitution is at amino acid position 1 of the fragment. In certain embodiments, the K132N substitution is at amino acid position 2 of the fragment. In certain embodiments, the K132N substitution is at amino acid position 3 of the fragment. In certain embodiments, the K132N substitution is at amino acid position 4 of the fragment. In certain embodiments, the K132N substitution is at amino acid position 5 of the fragment. In certain embodiments, the K132N substitution is at amino acid position 6 of the fragment. In certain embodiments, the K132N substitution is at amino acid position 7 of the fragment. In certain embodiments, the K132N substitution is at amino acid position 8 of the fragment. In certain embodiments, the K132N substitution is at amino acid position 9 of the fragment. In certain embodiments, the K132N substitution is at amino acid position 10 of the fragment. In preferred embodiments, the K132N substitution is at amino acid position 4 of the fragment. In preferred embodiments, the K132N substitution is at amino acid position 1 of the fragment

In certain embodiments, the C141Y substitution is at amino acid position 1 of the fragment. In certain embodiments, the C141Y substitution is at amino acid position 2 of the fragment. In certain embodiments, the C141Y substitution is at amino acid position 3 of the fragment. In certain embodiments, the C141Y substitution is at amino acid position 4 of the fragment. In certain embodiments, the C141Y substitution is at amino acid position 5 of the fragment. In certain embodiments, the C141Y substitution is at amino acid position 6 of the fragment. In certain embodiments, the C141Y substitution is at amino acid position 7 of the fragment. In certain embodiments, the C141Y substitution is at amino acid position 8 of the fragment. In certain embodiments, the C141Y substitution is at amino acid position 9 of the fragment. In certain embodiments, the C141Y substitution is at amino acid position 10 of the fragment. In preferred embodiments, the C141Y substitution is at amino acid position 3 of the fragment.

In certain embodiments, the P152L substitution is at amino acid position 1 of the fragment. In certain embodiments, the P152L substitution is at amino acid position 2 of the fragment. In certain embodiments, the P152L substitution is at amino acid position 3 of the fragment. In certain embodiments, the P152L substitution is at amino acid position 4 of the fragment. In certain embodiments, the P152L substitution is at amino acid position 5 of the fragment. In certain embodiments, the P152L substitution is at amino acid position 6 of the fragment. In certain embodiments, the P152L substitution is at amino acid position 7 of the fragment. In certain embodiments, the P152L substitution is at amino acid position 8 of the fragment. In certain embodiments, the P152L substitution is at amino acid position 9 of the fragment. In certain embodiments, the P152L substitution is at amino acid position 10 of the fragment. In preferred embodiments, the P152L substitution is at amino acid position 9 of the fragment.

In certain embodiments, the H193L substitution is at amino acid position 1 of the fragment. In certain embodiments, the H193L substitution is at amino acid position 2 of the fragment. In certain embodiments, the H193L substitution is at amino acid position 3 of the fragment. In certain embodiments, the H193L substitution is at amino acid position 4 of the fragment. In certain embodiments, the H193L substitution is at amino acid position 5 of the fragment. In certain embodiments, the H193L substitution is at amino acid position 6 of the fragment. In certain embodiments, the H193L substitution is at amino acid position 7 of the fragment. In certain embodiments, the H193L substitution is at amino acid position 8 of the fragment. In certain embodiments, the H193L substitution is at amino acid position 9 of the fragment. In certain embodiments, the H193L substitution is at amino acid position 10 of the fragment. In preferred embodiments, the H193L substitution is at amino acid position 7 of the fragment.

In certain embodiments, the H193Y substitution is at amino acid position 1 of the fragment. In certain embodiments, the H193Y substitution is at amino acid position 2 of the fragment. In certain embodiments, the H193Y substitution is at amino acid position 3 of the fragment. In certain embodiments, the H193Y substitution is at amino acid position 4 of the fragment. In certain embodiments, the H193Y substitution is at amino acid position 5 of the fragment. In certain embodiments, the H193Y substitution is at amino acid position 6 of the fragment. In certain embodiments, the H193Y substitution is at amino acid position 7 of the fragment. In certain embodiments, the H193Y substitution is at amino acid position 8 of the fragment. In certain embodiments, the H193Y substitution is at amino acid position 9 of the fragment. In certain embodiments, the H193Y substitution is at amino acid position 10 of the fragment. In preferred embodiments, the H193Y substitution is at amino acid position 7 of the fragment.

In certain embodiments, the Y220C substitution is at amino acid position 1 of the fragment. In certain embodiments, the Y220C substitution is at amino acid position 2 of the fragment. In certain embodiments, the Y220C substitution is at amino acid position 3 of the fragment. In certain embodiments, the Y220C substitution is at amino acid position 4 of the fragment. In certain embodiments, the Y220C substitution is at amino acid position 5 of the fragment. In certain embodiments, the Y220C substitution is at amino acid position 6 of the fragment. In certain embodiments, the Y220C substitution is at amino acid position 7 of the fragment. In certain embodiments, the Y220C substitution is at amino acid position 8 of the fragment. In certain embodiments, the Y220C substitution is at amino acid position 9 of the fragment. In certain embodiments, the Y220C substitution is at amino acid position 10 of the fragment. In preferred embodiments, the Y220C substitution is at amino acid position 4 of the fragment.

In certain embodiments, the V272M substitution is at amino acid position 1 of the fragment. In certain embodiments, the V272M substitution is at amino acid position 2 of the fragment. In certain embodiments, the V272M substitution is at amino acid position 3 of the fragment. In certain embodiments, the V272M substitution is at amino acid position 4 of the fragment. In certain embodiments, the V272M substitution is at amino acid position 5 of the fragment. In certain embodiments, the V272M substitution is at amino acid position 6 of the fragment. In certain embodiments, the V272M substitution is at amino acid position 7 of the fragment. In certain embodiments, the V272M substitution is at amino acid position 8 of the fragment. In certain embodiments, the V272M substitution is at amino acid position 9 of the fragment. In certain embodiments, the V272M substitution is at amino acid position 10 of the fragment. In preferred embodiments, the V272M substitution is at amino acid position 9 of the fragment.

In still further embodiments, the TP53 polypeptide fragment is selected from the group consisting of SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 94, SEQ ID NO: 95, and SEQ ID NO: 96. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 57 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 57. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 58 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 58. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 59 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 59. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 60 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 60. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 61 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 61. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 62 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 62. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 63 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 63. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 64 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 64. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 66 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 66. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 67 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 67. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 68 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 68. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 70 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 70. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 71 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 71. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 72 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 72. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 73 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 73. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 74 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 74. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 75 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 75. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 76 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 76. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 78 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 78. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 79 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 79. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 94 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 94. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 95 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 95. In certain embodiments, the TP53 polypeptide fragment is SEQ ID NO: 96 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 96.

In some embodiments, the TP53 polypeptide fragment has the sequence GX₂APPQYLX₉ (SEQ ID NO: 236), wherein X₂ is any amino acid other than leucine, and preferably methionine, and X₉ is any amino acid other than isoleucine, and preferably valine.

In some embodiments, the TP53 polypeptide fragment has the sequence AX₂NNMFCQX₉ (SEQ ID NO: 237), wherein X₂ is any amino acid other than leucine, and preferably methionine, and X₉ is any amino acid other than leucine, and preferably valine.

In some embodiments, the TP53 polypeptide fragment has the sequence NX₂FCQLAKX₉ (SEQ ID NO: 238), wherein X₂ is any amino acid other than methionine, and preferably leucine, and X₉ is any amino acid other than threonine, and preferably valine.

In some embodiments, the TP53 polypeptide fragment has the sequence QLWVDSTPX₉ (SEQ ID NO: 239), wherein X₉ is any amino acid other than leucine, and preferably isoleucine or valine.

In some embodiments, the TP53 polypeptide fragment has the sequence RLILTIITX₉ (SEQ ID NO: 240), wherein X₉ is any amino acid other than leucine, and preferably valine.

In some embodiments, the TP53 polypeptide fragment has the sequence YQGSYGFLX₉ (SEQ ID NO: 241), wherein X₉ is any amino acid other than leucine, and preferably isoleucine or valine.

In some embodiments, the TP53 polypeptide fragment has the sequence SX₂TCTYFPX₉ (SEQ ID NO: 242), wherein X₂ is any amino acid other than valine, and preferably leucine or methionine, and X₉ is any amino acid other than alanine, and preferably leucine, isoleucine, or valine.

In some embodiments, the TP53 polypeptide fragment has the sequence VX₂PCEPPEV (SEQ ID NO: 243), wherein X₂ is any amino acid other than valine, and preferably leucine or methionine.

In some embodiments, the TP53 polypeptide fragment has the sequence KX₂YPVQLWX₉ (SEQ ID NO: 244); wherein X₂ is any amino acid other than threonine, and preferably leucine or methionine, and X₉ is any amino acid other than valine, and preferably leucine or isoleucine.

In some embodiments, the TP53 polypeptide fragment has the sequence GX₂APPQLLX₉ (SEQ ID NO: 245), wherein X₂ is any amino acid other than leucine, and preferably methionine, and X₉ is any amino acid other than isoleucine, and preferably valine.

In some embodiments, the TP53 polypeptide fragment has the sequence LLGRNSFEX₉ (SEQ ID NO: 246), wherein X₉ is any amino acid other than methionine, and preferably leucine or isoleucine.

ERBB2 Polypeptides

Described herein are ERBB2 polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 104, and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length. Described herein are ERBB2 polypeptide fragments comprising, consisting of, or consisting essentially of an amino acid modification relative to SEQ ID NO: 104, and an amino acid substitution at amino acid position 3 of the fragment, amino acid position 10 of the fragment, or both, wherein the fragment is at least ten amino acids in length.

In certain embodiments, the modification comprises a deletion, insertion, and/or substitution. In preferred embodiments, the modification comprises a substitution. In further preferred embodiments, the modification comprises valine to isoleucine amino acid substitution at a position corresponding to position 842 of SEQ ID NO: 104 (V842I).

Described herein are ERBB2 polypeptide fragments comprising, consisting of, or consisting essentially of a valine to isoleucine amino acid substitution at a position corresponding to position 842 of SEQ ID NO: 104 (V842I), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is nine amino acids in length. In certain embodiments, the ERBB2 polypeptide fragments comprise, consist of, or consist essentially of a valine to isoleucine amino acid substitution at a position corresponding to position 842 of SEQ ID NO: 104 (V842I), and an amino acid substitution at amino acid position 2 of the fragment. In certain embodiments, the ERBB2 polypeptide fragments comprise, consist of, or consist essentially of a valine to isoleucine amino acid substitution at a position corresponding to position 842 of SEQ ID NO: 104 (V842I), and an amino acid substitution at amino acid position 9 of the fragment. In certain embodiments, the ERBB2 polypeptide fragments comprise, consist of, or consist essentially of a valine to isoleucine amino acid substitution at a position corresponding to position 842 of SEQ ID NO: 104 (V842I), and an amino acid substitution at amino acid position 2 of the fragment and at position 9 of the fragment.

In certain embodiments, the ERBB2 polypeptide fragment binds to HLA 2.1 (HLA-A*02:01), HLA-A*01:01, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*33:03, HLA-C*07:01, HLA-C*07:02, HLA-C*04:01, HLA-B*07:02, HLA-B*44:02, or HLA-B*35:01. In preferred embodiments, the ERBB2 polypeptide fragment binds to HLA-A*02:01.

In some embodiments, the ERBB2 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment. In certain embodiments, binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control, expressed as a percent (%) binding affinity. In certain embodiments, the ERBB2 polypeptide fragment has a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment, wherein binding affinity for HLA-A*02:01 is a measurement of average relative binding to a positive 9-mer polypeptide control.

In certain embodiments, the V8421 substitution is at amino acid position 1 of the fragment. In certain embodiments, the V8421 substitution is at amino acid position 2 of the fragment. In certain embodiments, the V8421 substitution is at amino acid position 3 of the fragment. In certain embodiments, the V8421 substitution is at amino acid position 4 of the fragment. In certain embodiments, the V8421 substitution is at amino acid position 5 of the fragment. In certain embodiments, the V8421 substitution is at amino acid position 6 of the fragment. In certain embodiments, the V8421 substitution is at amino acid position 7 of the fragment. In certain embodiments, the V8421 substitution is at amino acid position 8 of the fragment. In certain embodiments, the V8421 substitution is at amino acid position 9 of the fragment. In certain embodiments, the V8421 substitution is at amino acid position 10 of the fragment. In preferred embodiments, the V8421 substitution at amino acid position 3 of the fragment.

In still further embodiments, the ERBB2 polypeptide fragment is selected from the group consisting of SEQ ID NO: 84, SEQ ID NO: 85, and SEQ ID NO: 86. In certain embodiments, the ERBB2 polypeptide fragment is SEQ ID NO: 84 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 84. In certain embodiments, the ERBB2 polypeptide fragment is SEQ ID NO: 85 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 85. In certain embodiments, the ERBB2 polypeptide fragment is SEQ ID NO: 86 or a sequence having at least 90% sequence identity, at least 95% sequence identity or at least 99% sequence identity to SEQ ID NO: 86.

In some embodiments, the ERBB2 polypeptide fragment has the sequence RX₂IHRDLAX₉ (SEQ ID NO: 247), wherein X₂ is any amino acid other than leucine, and preferably methionine, and X₉ is any amino acid other than alanine, and preferably leucine or valine.

T-Cell Receptors

TCRs may be generated that bind the polypeptide fragments of the disclosure. The TCRs may be identified based on T-cell binding to the polypeptide fragments, followed by sequencing of the TCR. The identified TCR may be identified from αβ T cells. The identified TCRs may be further engineered to improve their affinity, stability, solubility or the like. For example, TCRs may be cysteine stabilized, expressed as soluble TCRs, as single chain TCRs, as fusion with N-terminal or C-terminal epitope tags, engineered to improve stability with mutations in hydrophobic core, such as positions 11, 13, 19, 21, 53, 76, 89, 91 or 94 of the α chain, domain swapped with α and β chain variable and/or constant domains swapped as described in U.S. Pat. No. 7,329,731, U.S. Pat. No. 7,569,664, U.S. Pat. No. 9,133,264, U.S. Pat. No. 9,624,292, US2016/0130319 and U.S. Pat. No. 9,884,075.

Described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18, and (b) the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 14, Table 15, Table 16, Table 17, or Table 18. An alpha chain CDR3 corresponds to a beta chain CDR3 if they appear in the same row in Table 14, Table 15, Table 16, Table 17 or Table 18.

Also described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, a CDR2 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, and a CDR3 comprising an amino acid sequence provided in Table 14, Table 15, Table 16, Table 17, or Table 18, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, a CDR2 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, and a CDR3 comprising a corresponding amino acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18. An alpha chain CDR1 or CDR2 corresponds to a beta chain CDR1 or CDR2 if they appear in the same row in Table 19, Table 20, Table 21, Table 22, or Table 23. An alpha chain CDR3 corresponds to a beta chain CDR3 if they appear in the same row in Table 14, Table 15, Table 16, Table 17, or Table 18. An alpha and beta chain CDR1 and CDR2 provided in Table 19, Table 20, Table 21, Table 22, or Table 23 correspond to an alpha and beta chain CDR3 provided in the same row in Table 14, Table 15, Table 16, Table 17 or Table 18.

Described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 14, and (b) the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 14. In certain embodiments, the TCRs provided in Table 14 recognize the PIK3CA mutant-mimic fragments SEQ ID NO: 9 and SEQ ID NO: 45.

Also described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 19, a CDR2 comprising an amino acid sequence provided in Table 19, and a CDR3 comprising an amino acid sequence provided in Table 14, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 19, a CDR2 comprising an amino acid sequence provided in Table 19, and a CDR3 comprising a corresponding amino acid sequence provided in Table 14. In certain embodiments, the TCRs provided in Table 19 and Table 14 recognize the PIK3CA mutant-mimic fragments SEQ ID NO: 9 and SEQ ID NO: 45. An alpha and beta chain CDR1 and CDR2 provided in Table 19 correspond to and alpha and beta chain CDR3 provided in the same row in Table 14.

In certain embodiments, described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 120 or having at least 90% sequence identity to SEQ ID NO: 120 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (b) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 118 or having at least 90% sequence identity to SEQ ID NO: 118 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (c) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 122 or having at least 90% sequence identity to SEQ ID NO: 122 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (d) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 122 or having at least 90% sequence identity to SEQ ID NO: 122 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 124 or having at least 90% sequence identity to SEQ ID NO: 124; (e) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 122 or having at least 90% sequence identity to SEQ ID NO: 122 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 134 or having at least 90% sequence identity to SEQ ID NO: 134; (f) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 112 or having at least 90% sequence identity to SEQ ID NO: 112 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (g) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 116 or having at least 90% sequence identity to SEQ ID NO: 116 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (h) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 116 or having at least 90% sequence identity to SEQ ID NO: 116 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 128 or having at least 90% sequence identity to SEQ ID NO: 128; (i) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 126 or having at least 90% sequence identity to SEQ ID NO: 126 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (j) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 or having at least 90% sequence identity to SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (k) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 or having at least 90% sequence identity to SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 132 or having at least 90% sequence identity to SEQ ID NO: 132; or (l) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 or having at least 90% sequence identity to SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 134 or having at least 90% sequence identity to SEQ ID NO: 134.

Described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 15, and the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 15. In certain embodiments, the TCRs provided in Table 15 recognize the TP53 mutant-mimic fragments SEQ ID NO: 13 and SEQ ID NO: 59.

Also described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 20, a CDR2 comprising an amino acid sequence provided in Table 20, and a CDR3 comprising an amino acid sequence provided in Table 15, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 20, a CDR2 comprising an amino acid sequence provided in Table 20, and a CDR3 comprising a corresponding amino acid sequence provided in Table 15. In certain embodiments, the TCRs provided in Table 20 and Table 15 recognize the TP53 mutant-mimic fragments SEQ ID NO: 13 and SEQ ID NO: 59. An alpha and beta chain CDR1 and CDR2 provided in Table 20 correspond to and alpha and beta chain CDR3 provided in the same row in Table 15.

In certain embodiments, described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 118 or having at least 90% sequence identity to SEQ ID NO: 118 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (b) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 118 or having at least 90% sequence identity to SEQ ID NO: 118 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 142 or having at least 90% sequence identity to SEQ ID NO: 142; (c) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 207 or having at least 90% sequence identity to SEQ ID NO: 207 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (d) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 112 or having at least 90% sequence identity to SEQ ID NO: 112 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (e) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 205 or having at least 90% sequence identity to SEQ ID NO: 205 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (f) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 172 or having at least 90% sequence identity to SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (g) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 172 or having at least 90% sequence identity to SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 166 or having at least 90% sequence identity to SEQ ID NO: 166; (h) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 186 or having at least 90% sequence identity to SEQ ID NO: 186 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (i) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 152 or having at least 90% sequence identity to SEQ ID NO: 152 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (j) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 or having at least 90% sequence identity to SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (k) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 or having at least 90% sequence identity to SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 142 or having at least 90% sequence identity to SEQ ID NO: 142; (l) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 or having at least 90% sequence identity to SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 150 or having at least 90% sequence identity to SEQ ID NO: 150; (m) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 or having at least 90% sequence identity to SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 162 or having at least 90% sequence identity to SEQ ID NO: 162; (n) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 136 or having at least 90% sequence identity to SEQ ID NO: 136 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (o) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 136 or having at least 90% sequence identity to SEQ ID NO: 136 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 138 or having at least 90% sequence identity to SEQ ID NO: 138; (p) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 136 or having at least 90% sequence identity to SEQ ID NO: 136 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 142 or having at least 90% sequence identity to SEQ ID NO: 142; (q) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 140 or having at least 90% sequence identity to SEQ ID NO: 140 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (r) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 140 or having at least 90% sequence identity to SEQ ID NO: 140 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 160 or having at least 90% sequence identity to SEQ ID NO: 160; (s) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 144 or having at least 90% sequence identity to SEQ ID NO: 144 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (t) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 144 or having at least 90% sequence identity to SEQ ID NO: 144 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 146 or having at least 90% sequence identity to SEQ ID NO: 146; (u) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 144 or having at least 90% sequence identity to SEQ ID NO: 144 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 158 or having at least 90% sequence identity to SEQ ID NO: 158; (v) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 148 or having at least 90% sequence identity to SEQ ID NO: 148 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (w) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 148 or having at least 90% sequence identity to SEQ ID NO: 148 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 150 or having at least 90% sequence identity to SEQ ID NO: 150; (x) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 154 or having at least 90% sequence identity to SEQ ID NO: 154 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 156 or having at least 90% sequence identity to SEQ ID NO: 156; (y) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 or having at least 90% sequence identity to SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (z) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 or having at least 90% sequence identity to SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 166 or having at least 90% sequence identity to SEQ ID NO: 166; (aa) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 or having at least 90% sequence identity to SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 180 or having at least 90% sequence identity to SEQ ID NO: 180; (bb) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 or having at least 90% sequence identity to SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 182 or having at least 90% sequence identity to SEQ ID NO: 182; (cc) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 or having at least 90% sequence identity to SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 197 or having at least 90% sequence identity to SEQ ID NO: 197; (dd) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 168 or having at least 90% sequence identity to SEQ ID NO: 168 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (ee) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 168 or having at least 90% sequence identity to SEQ ID NO: 168 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 170 or having at least 90% sequence identity to SEQ ID NO: 170; (ff) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 168 or having at least 90% sequence identity to SEQ ID NO: 168 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 199 or having at least 90% sequence identity to SEQ ID NO: 199; (gg) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 174 or having at least 90% sequence identity to SEQ ID NO: 174 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 176 or having at least 90% sequence identity to SEQ ID NO: 176; (hh) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 174 or having at least 90% sequence identity to SEQ ID NO: 174 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 178 or having at least 90% sequence identity to SEQ ID NO: 178; (ii) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 184 or having at least 90% sequence identity to SEQ ID NO: 184 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (jj) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 184 or having at least 90% sequence identity to SEQ ID NO: 184 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 188 or having at least 90% sequence identity to SEQ ID NO: 188; (kk) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 190 or having at least 90% sequence identity to SEQ ID NO: 190 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 192 or having at least 90% sequence identity to SEQ ID NO: 192; (ll) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 194 or having at least 90% sequence identity to SEQ ID NO: 194 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (mm) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 201 or having at least 90% sequence identity to SEQ ID NO: 201 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 203 or having at least 90% sequence identity to SEQ ID NO: 203; or (nn) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 210 or having at least 90% sequence identity to SEQ ID NO: 210 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114.

Described herein are T-cell receptors (TCRs) comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a complementarity determining region 3 (CDR3) comprising an amino acid sequence provided in Table 16, and the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 16. In certain embodiments, the TCRs provided in Table 16 recognize the TP53 mutant-mimic fragments SEQ ID NO: 18 and SEQ ID NO: 68.

Also described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 21, a CDR2 comprising an amino acid sequence provided in Table 21, and a CDR3 comprising an amino acid sequence provided in Table 16, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 21, a CDR2 comprising an amino acid sequence provided in Table 21, and a CDR3 comprising a corresponding amino acid sequence provided in Table 16. In certain embodiments, the TCRs provided in Table 21 and Table 16 recognize the TP53 mutant-mimic fragments SEQ ID NO: 18 and SEQ ID NO: 68. An alpha and beta chain CDR1 and CDR2 provided in Table 21 correspond to and alpha and beta chain CDR3 provided in the same row in Table 16.

In certain embodiments, described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 172 or having at least 90% sequence identity to SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (b) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 172 or having at least 90% sequence identity to SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 170 or having at least 90% sequence identity to SEQ ID NO: 170; (c) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 172 or having at least 90% sequence identity to SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 213 or having at least 90% sequence identity to SEQ ID NO: 213; (d) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 152 or having at least 90% sequence identity to SEQ ID NO: 152 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (e) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 168 or having at least 90% sequence identity to SEQ ID NO: 168 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 170 or having at least 90% sequence identity to SEQ ID NO: 170; (f) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 215 or having at least 90% sequence identity to SEQ ID NO: 215 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (g) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 217 or having at least 90% sequence identity to SEQ ID NO: 217 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (h) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 219 or having at least 90% sequence identity to SEQ ID NO: 219 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (i) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 219 or having at least 90% sequence identity to SEQ ID NO: 219 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 221 or having at least 90% sequence identity to SEQ ID NO: 221; or (j) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 223 or having at least 90% sequence identity to SEQ ID NO: 223 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 221 or having at least 90% sequence identity to SEQ ID NO: 221.

Described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 17, and the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 17. In certain embodiments, the TCRs provided in Table 17 recognize the CTNNB1 mutant-mimic fragments SEQ ID NO: 3 and SEQ ID NO: 32.

Also described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 22, a CDR2 comprising an amino acid sequence provided in Table 22, and a CDR3 comprising an amino acid sequence provided in Table 17, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 22, a CDR2 comprising an amino acid sequence provided in Table 22, and a CDR3 comprising a corresponding amino acid sequence provided in Table 17. In certain embodiments, the TCRs provided in Table 22 and Table 17 recognize the CTNNB1 mutant-mimic fragments SEQ ID NO: 3 and SEQ ID NO: 32. An alpha and beta chain CDR1 and CDR2 provided in Table 22 correspond to and alpha and beta chain CDR3 provided in the same row in Table 17.

In certain embodiments, described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 152 or having at least 90% sequence identity to SEQ ID NO: 152 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 252 or having at least 90% sequence identity to SEQ ID NO: 252; (b) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 152 or having at least 90% sequence identity to SEQ ID NO: 152 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (c) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 248 or having at least 90% sequence identity to SEQ ID NO: 248 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 250 or having at least 90% sequence identity to SEQ ID NO: 250; (d) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 248 or having at least 90% sequence identity to SEQ ID NO: 248 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 258 or having at least 90% sequence identity to SEQ ID NO: 258; (e) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 254 or having at least 90% sequence identity to SEQ ID NO: 254 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 256 or having at least 90% sequence identity to SEQ ID NO: 256; (f) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 254 or having at least 90% sequence identity to SEQ ID NO: 254 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (g) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 263 or having at least 90% sequence identity to SEQ ID NO: 263 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 265 or having at least 90% sequence identity to SEQ ID NO: 265; (h) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 267 or having at least 90% sequence identity to SEQ ID NO: 267 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 269 or having at least 90% sequence identity to SEQ ID NO: 269; (i) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 267 or having at least 90% sequence identity to SEQ ID NO: 267 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 271 or having at least 90% sequence identity to SEQ ID NO: 271; or (j) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 273 or having at least 90% sequence identity to SEQ ID NO: 273 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 275 or having at least 90% sequence identity to SEQ ID NO: 275; or (k) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 277 or having at least 90% sequence identity to SEQ ID NO: 277 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 279 or having at least 90% sequence identity to SEQ ID NO: 279; or (l) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 281 or having at least 90% sequence identity to SEQ ID NO: 281 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 283 or having at least 90% sequence identity to SEQ ID NO: 283; or (m) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 285 or having at least 90% sequence identity to SEQ ID NO: 285 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 287 or having at least 90% sequence identity to SEQ ID NO: 287.

Described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 18, and the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 18. In certain embodiments, the TCRs provided in Table 18 recognize the TP53 mutant-mimic fragments SEQ ID NO: 23 and SEQ ID NO: 78.

Also described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 23, a CDR2 comprising an amino acid sequence provided in Table 23, and a CDR3 comprising an amino acid sequence provided in Table 18, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 23, a CDR2 comprising an amino acid sequence provided in Table 23, and a CDR3 comprising a corresponding amino acid sequence provided in Table 18. In certain embodiments, the TCRs provided in Table 23 and Table 18 recognize the TP53 mutant-mimic fragments SEQ ID NO: 23 and SEQ ID NO: 78. An alpha and beta chain CDR1 and CDR2 provided in Table 23 correspond to and alpha and beta chain CDR3 provided in the same row in Table 18.

In certain embodiments, described herein are TCRs comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 248 or having at least 90% sequence identity to SEQ ID NO: 248 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (b) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 289 or having at least 90% sequence identity to SEQ ID NO: 289 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114 or having at least 90% sequence identity to SEQ ID NO: 114; (c) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 289 or having at least 90% sequence identity to SEQ ID NO: 289 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 291 or having at least 90% sequence identity to SEQ ID NO: 291.

Polynucleotides

The disclosure also provides polynucleotides that encode any of the polypeptide fragments or TCRs disclosed herein.

In some embodiments, the polynucleotide encodes a polypeptide fragment selected from the group consisting of SEQ ID NO: 25 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 25, SEQ ID NO: 26 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 26, SEQ ID NO: 27 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 27, SEQ ID NO: 28 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 28, SEQ ID NO: 29 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 29, SEQ ID NO: 30 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 30, SEQ ID NO: 31 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 31, SEQ ID NO: 32 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 32, SEQ ID NO: 33 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 33, SEQ ID NO: 37 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 37, SEQ ID NO: 38 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 38, SEQ ID NO: 39 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 39, SEQ ID NO: 40 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 40, SEQ ID NO: 41 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 41, SEQ ID NO: 42 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 42, SEQ ID NO: 43 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 43, SEQ ID NO: 44 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 44, SEQ ID NO: 45 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 45, SEQ ID NO: 46 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 46, SEQ ID NO: 47 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 47, SEQ ID NO: 48 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 48, SEQ ID NO: 49 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 49, SEQ ID NO: 51 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 51, SEQ ID NO: 52 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 52, SEQ ID NO: 53 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 53, SEQ ID NO: 54 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 54, SEQ ID NO: 55 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 55, SEQ ID NO: 56 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 56, SEQ ID NO: 57 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 57, SEQ ID NO: 58 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 58, SEQ ID NO: 59 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 59, SEQ ID NO: 60 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 60, SEQ ID NO: 61 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 61, SEQ ID NO: 62 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 62, SEQ ID NO: 63 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 63, SEQ ID NO: 64 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 64, SEQ ID NO: 66 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 66, SEQ ID NO: 67 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 67, SEQ ID NO: 68 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 68, SEQ ID NO: 70 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 70, SEQ ID NO: 71 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 71, SEQ ID NO: 72 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 72, SEQ ID NO: 73 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 73, SEQ ID NO: 74 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 74, SEQ ID NO: 75 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 75, SEQ ID NO: 76 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 76, SEQ ID NO: 78 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 78, SEQ ID NO: 79 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 79, SEQ ID NO: 80 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 80, SEQ ID NO: 81 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 81, SEQ ID NO: 84 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 84, SEQ ID NO: 85 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 85, SEQ ID NO: 86 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 86, SEQ ID NO: 88 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 88, SEQ ID NO: 90 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 90, SEQ ID NO: 91 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 91, SEQ ID NO: 92 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 92, SEQ ID NO: 93 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 93, SEQ ID NO: 94 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 94, SEQ ID NO: 95 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 95, SEQ ID NO: 96 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 96, and any combination thereof.

In some embodiments, the polynucleotide encodes a polypeptide fragment selected from the group consisting of SEQ ID NO: 29 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 29, SEQ ID NO: 32 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 32, SEQ ID NO: 45 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 45, SEQ ID NO: 59 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 59, SEQ ID NO: 64 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 64, SEQ ID NO: 68 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 68, SEQ ID NO: 75 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 75, SEQ ID NO: 78 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 78, and any combination thereof.

In some embodiments, the polynucleotide encodes a TCR polypeptide comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a complementarity determining region 3 (CDR3) comprising an amino acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18, and the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 14, Table 15, Table 16, Table 17, or Table 18.

In some embodiments, the polynucleotide encodes a TCR polypeptide comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a complementarity determining region 3 (CDR3) that is encoded by a nucleic acid sequence provided in Table 14, Table 15, Table 16, Table 17, or Table 18, and the beta chain comprises a CDR3 that is encoded by a corresponding nucleic acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18.

In some embodiments, the polynucleotide comprises DNA.

In some embodiments, the polynucleotide comprises RNA.

In some embodiments, RNA is mRNA.

In some embodiments, the polynucleotide comprises a promoter, an enhancer, a polyadenylation site, a Kozak sequence, a stop codon, or any combination thereof.

Methods of generating polynucleotides of the disclosure are known in the art and include chemical synthesis, enzymatic synthesis (e.g. in vitro transcription), enzymatic or chemical cleavage of a longer precursor, chemical synthesis of smaller fragments of the polynucleotides followed by ligation of the fragments or known PCR methods. The polynucleotide sequence to be synthesized may be designed with the appropriate codons for the desired amino acid sequence. In general, preferred codons may be selected for the intended host in which the sequence will be used for expression

Vectors

The disclosure also provides vectors comprising any of the polynucleotides disclosed herein. The disclosure also provides vectors comprising a polynucleotide encoding for any of the polypeptides disclosed herein.

In some embodiments, vector comprises a polynucleotide that encodes a polypeptide fragment selected from the group consisting of SEQ ID NO: 25 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 25, SEQ ID NO: 26 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 26, SEQ ID NO: 27 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 27, SEQ ID NO: 28 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 28, SEQ ID NO: 29 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 29, SEQ ID NO: 30 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 30, SEQ ID NO: 31 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 31, SEQ ID NO: 32 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 32, SEQ ID NO: 33 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 33, SEQ ID NO: 37 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 37, SEQ ID NO: 38 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 38, SEQ ID NO: 39 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 39, SEQ ID NO: 40 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 40, SEQ ID NO: 41 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 41, SEQ ID NO: 42 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 42, SEQ ID NO: 43 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 43, SEQ ID NO: 44 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 44, SEQ ID NO: 45 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 45, SEQ ID NO: 46 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 46, SEQ ID NO: 47 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 47, SEQ ID NO: 48 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 48, SEQ ID NO: 49 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 49, SEQ ID NO: 51 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 51, SEQ ID NO: 52 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 52, SEQ ID NO: 53 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 53, SEQ ID NO: 54 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 54, SEQ ID NO: 55 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 55, SEQ ID NO: 56 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 56, SEQ ID NO: 57 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 57, SEQ ID NO: 58 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 58, SEQ ID NO: 59 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 59, SEQ ID NO: 60 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 60, SEQ ID NO: 61 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 61, SEQ ID NO: 62 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 62, SEQ ID NO: 63 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 63, SEQ ID NO: 64 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 64, SEQ ID NO: 66 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 66, SEQ ID NO: 67 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 67, SEQ ID NO: 68 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 68, SEQ ID NO: 70 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 70, SEQ ID NO: 71 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 71, SEQ ID NO: 72 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 72, SEQ ID NO: 73 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 73, SEQ ID NO: 74 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 74, SEQ ID NO: 75 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 75, SEQ ID NO: 76 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 76, SEQ ID NO: 78 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 78, SEQ ID NO: 79 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 79, SEQ ID NO: 80 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 80, SEQ ID NO: 81 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 81, SEQ ID NO: 84 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 84, SEQ ID NO: 85 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 85, SEQ ID NO: 86 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 86, SEQ ID NO: 88 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 88, SEQ ID NO: 90 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 90, SEQ ID NO: 91 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 91, SEQ ID NO: 92 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 92, SEQ ID NO: 93 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 93, SEQ ID NO: 94 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 94, SEQ ID NO: 95 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 95, SEQ ID NO: 96 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 96, and any combination thereof.

In some embodiments, vector comprises a polynucleotide that encodes a polypeptide fragment selected from the group consisting of SEQ ID NO: 29 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 29, SEQ ID NO: 32 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 32, SEQ ID NO: 45 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 45, SEQ ID NO: 59 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 59, SEQ ID NO: 64 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 64, SEQ ID NO: 68 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 68, SEQ ID NO: 75 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 75, SEQ ID NO: 78 or a sequence having at least 90% identity, at least 95% identity, or at least 99% identity to SEQ ID NO: 78, and any combination thereof.

In some embodiments, the vector comprises a polynucleotide that encodes a TCR polypeptide comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a complementarity determining region 3 (CDR3) comprising an amino acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18, and the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18.

In some embodiments, the vector comprises a polynucleotide that encodes a TCR polypeptide comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, a CDR2 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, and a CDR3 comprising an amino acid sequence provided in Table 14, Table 15, Table 16, Table 17, or Table 18, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, a CDR2 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, and a CDR3 comprising a corresponding amino acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18. An alpha chain CDR1 or CDR2 corresponds to a beta chain CDR1 or CDR2 if they appear in the same row in Table 19, Table 20, Table 21, Table 22, or Table 23. An alpha chain CDR3 corresponds to a beta chain CDR3 if they appear in the same row in Table 14, Table 15, Table 16, Table 17, or Table 18. An alpha and beta chain CDR1 and CDR2 provided in Table 19, Table 20, Table 21, Table 22, or Table 23 correspond to an alpha and beta chain CDR3 provided in the same row in Table 14, Table 15, Table 16, Table 17 or Table 18.

In some embodiments, the vector comprises a polynucleotide that encodes a TCR polypeptide comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a complementarily determining region 3 (CDR3) that is encoded by a nucleic acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18, and the beta chain comprises a CDR3 that is encoded by a corresponding nucleic acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18.

In some embodiments, the vector comprises a polynucleotide that encodes a TCR polypeptide comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 that is encoded by a nucleic acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, a CDR2 that is encoded by a nucleic acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, and a CDR3 that is encoded by a nucleic acid sequence provided in Table 14, Table 15, Table 16, Table 17, or Table 18, and (b) the beta chain comprises a CDR1 that is encoded by a nucleic acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, a CDR2 that is encoded by a nucleic acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, and a CDR3 that is encoded by a nucleic acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18. An alpha chain CDR1 or CDR2 corresponds to a beta chain CDR1 or CDR2 if they appear in the same row in Table 19, Table 20, Table 21, Table 22, or Table 23. An alpha chain CDR3 corresponds to a beta chain CDR3 if they appear in the same row in Table 14, Table 15, Table 16, Table 17, or Table 18. An alpha and beta chain CDR1 and CDR2 provided in Table 19, Table 20, Table 21, Table 22, or Table 23 correspond to and alpha and beta chain CDR3 provided in the same row in Table 14, Table 15, Table 16, Table 17 or Table 18.

The vector may be a vector intended for expression of the polynucleotide of the disclosure in any host, such as bacteria, yeast or a mammal. Suitable expression vectors are typically replicable in the host organisms either as episomes or as an integral part of the host chromosomal DNA. Commonly, expression vectors contain selection markers such as ampicillin-resistance, hygromycin-resistance, tetracycline resistance, kanamycin resistance or neomycin resistance to permit detection of those cells transformed or transduced with the desired DNA sequences. Exemplary vectors are plasmids, cosmids, phages, viral vectors or artificial chromosomes.

Suitable vectors that may be used are—Bacterial: pBs, phagescript, PsiX174, pBluescript SK, pBs KS, pNH8a, pNH16a, pNH18a, pNH46a (Stratagene, La Jolla, Calif, USA); pTrc99A, pKK223-3, pKK233-3, pDR540, and pRIT5 (Pharmacia, Uppsala, Sweden). Eukaryotic: pWLneo, pSV2cat, pOG44, PXR1, pSG (Stratagene), pSVK3, pBPV, pMSG and pSVL (Pharmacia).

The disclosure provides an expression vector comprising the polynucleotide of the disclosure. The disclosure also provides an expression vector comprising the polynucleotide encoding for the polypeptide of the disclosure.

Other Viral Vectors and Recombinant Viruses

The disclosure also provides a viral vector comprising any of the polynucleotides of the disclosure.

The disclosure also provides a viral vector comprising a polynucleotide encoding any of the polypeptides of the disclosure.

Viral vectors are derived from naturally occurring virus genomes, which typically are modified to be replication incompetent, e.g. non-replicating. Non-replicating viruses require the provision of proteins in trans for replication. Typically, those proteins are stably or transiently expressed in a viral producer cell line, thereby allowing replication of the virus. The viral vectors are, thus, typically infectious and non-replicating. Viral vectors may be adenovirus vectors, adeno-associated virus (AAV) vectors (e.g., AAV type 5 and type 2), Great ape adenovirus vectors (GAd), alphavirus vectors (e.g., Venezuelan equine encephalitis virus (VEE), Sindbis virus (SIN), Semliki forest virus (SFV), and VEE-SIN chimeras), herpes virus vectors (e.g. vectors derived from cytomegaloviruses, like rhesus cytomegalovirus (RhCMV)), arena virus vectors (e.g. lymphocytic choriomeningitis virus (LCMV) vectors), measles virus vectors, pox virus vectors (e.g., vaccinia virus, modified vaccinia virus Ankara (MVA), NYVAC (derived from the Copenhagen strain of vaccinia), and avipox vectors: canarypox (ALVAC) and fowlpox (FPV) vectors), vesicular stomatitis virus vectors, retrovirus vectors, lentivirus vectors, viral like particles, and bacterial spores.

In some embodiments, the viral vector is derived from adenovirus, poxvirus, alphavirus, adeno-associated virus, retrovirus, or a self-replicating RNA molecule.

In some embodiments, the viral vector is derived from an adenovirus. In certain embodiments, the adenovirus vector is selected from hAd5, hAd7, hAd11, hAd26, hAd34, hAd35, hAd48, hAd49, hAd50, GAd20, Gad19, GAd21, GAd25, GAd26, GAd27, GAd28, GAd29, GAd30, GAd31, ChAd3, ChAd4, ChAd5, ChAd6, ChAd7, ChAd8, ChAd9, ChAd10, ChAd11, ChAd16, ChAdI7, ChAd19, ChAd20, ChAd22, ChAd24, ChAd26, ChAd30, ChAd31, ChAd37, ChAd38, ChAd44, ChAd55, ChAd63, ChAd73, ChAd82, ChAd83, ChAd146, ChAd147, PanAd1, PanAd2, and PanAd3.

Adenovirus vectors may be derived from human adenovirus (Ad) but also from adenoviruses that infect other species, such as bovine adenovirus (e.g. bovine adenovirus 3, BAdV3), a canine adenovirus (e.g. CAdV2), a porcine adenovirus (e.g. PAdV3 or 5), or great apes, such as Chimpanzee (Pan), Gorilla (Gorilla), Orangutan (Pongo), Bonobo (Pan paniscus) and common chimpanzee (Pan troglodytes). Typically, naturally occurring great ape adenoviruses are isolated from stool samples of the respective great ape.

Human adenovirus vectors may be derived from various adenovirus serotypes, for example from human adenovirus serotypes hAd5, hAd7, hAd11, hAd26, hAd34, hAd35, hAd48, hAd49 or hAd50 (the serotypes are also referred to as Ad5, Ad7, Ad11, Ad26, Ad34, Ad35, Ad48, Ad49 or Ad50).

Great ape adenovirus (GAd) vectors may be derived from various adenovirus serotypes, for example from great ape adenovirus serotypes GAd20, Gad19, GAd21, GAd25, GAd26, GAd27, GAd28, GAd29, GAd30, GAd31, ChAd3, ChAd4, ChAd5, ChAd6, ChAd7, ChAd8, ChAd9, ChAd10, ChAd11, ChAd16, ChAdI7, ChAd19, ChAd20, ChAd22, ChAd24, ChAd26, ChAd30, ChAd31, ChAd37, ChAd38, ChAd44, ChAd55, ChAd63, ChAd73, ChAd82, ChAd83, ChAd146, ChAd147, PanAd1, PanAd2, or PanAd3.

Adenovirus vectors are known in the art. The sequences of most of the human and non-human adenoviruses are known, and for others can be obtained using routine procedures. An exemplary genome sequence of Ad26 is found in GenBank Accession number EF153474 and in Int. Pat. Publ. No. WO2007/104792. An exemplary genome sequence of Ad35 is found in Int. Pat. Publ. No. WO2000/70071. Vectors based on Ad26 are described for example, in Int. Pat. Publ. No. WO2007/104792. Vectors based on Ad35 are described for example in U.S. Pat. No. 7,270,811 and Int. Pat. Publ. No. WO2000/70071. Vectors based on ChAd3, ChAd4, ChAd5, ChAd6, ChAd7, ChAd8, ChAd9, ChAd10, ChAd11, ChAd16, ChAd17, ChAd19, ChAd20, ChAd22, ChAd24, ChAd26, ChAd30, ChAd31, ChAd37, ChAd38, ChAd44, ChAd63 and ChAd82 are described in WO2005/071093. Vectors based on PanAd1, PanAd2, PanAd3, ChAd55, ChAd73, ChAd83, ChAd146, and ChAd147 are described in Int. Pat. Publ. No. WO2010/086189.

In some embodiments, the viral vector is a poxvirus. In some embodiments, the poxvirus vector is selected from smallpox virus vector, vaccinia virus vector, cowpox virus vector, monkeypox virus vector, Copenhagen vaccinia virus (W) vector, New York Attenuated Vaccinia Virus (NYVAC) vector, and Modified Vaccinia Ankara (MVA) vector.

Poxvirus (Poxviridae) vectors may be derived from smallpox virus (variola), vaccinia virus, cowpox virus or monkeypox virus. Exemplary vaccinia viruses are the Copenhagen vaccinia virus (W), New York Attenuated Vaccinia Virus (NYVAC), ALVAC, TROVAC or Modified Vaccinia Ankara (MVA).

MVA originates from the dermal vaccinia strain Ankara (Chorioallantois vaccinia Ankara (CVA) virus) that was maintained in the Vaccination Institute, Ankara, Turkey for many years and used as the basis for vaccination of humans. However, due to the often severe post-vaccinal complications associated with vaccinia viruses (VACV), there were several attempts to generate a more attenuated, safer smallpox vaccine.

In some embodiments, the viral vector is an adeno-associated virus. The viral vector comprising the polynucleotides of the disclosure may be derived from human adeno-associated viruses, such as AAV-2 (adeno-associated virus type 2). An attractive feature of AAV vectors is that they do not express any viral genes. The only viral DNA sequences included in the AAV vectors are the 145 bp inverted terminal repeats (ITR). Thus, as in immunization with naked DNA, the only gene expressed is that of the antigen, or antigen chimera. Additionally, AAV vectors are known to transduce both dividing and non-dividing cells, such as human peripheral blood monocyte-derived dendritic cells, with persistent transgene expression, and with the possibility of oral and intranasal delivery for generation of mucosal immunity. Moreover, the amount of DNA required appears to be much less by several orders of magnitude, with maximum responses at doses of 10¹⁰ to 10ⁿ particles or copies of DNA in contrast to naked DNA doses of 50 μg or about 10¹⁵ copies. AAV vectors are packaged by co-transfection of a suitable cell line (e.g., human 293 cells) with the DNA contained in the AAV ITR chimeric protein encoding constructs and an AAV helper plasmid ACG2 containing the AAV coding region (AAV rep and cap genes) without the ITRs. The cells are subsequently infected with the adenovirus Ad5. Vectors can be purified from cell lysates using methods known in the art (e.g., such as cesium chloride density gradient ultracentrifugation) and are validated to ensure that they are free of detectable replication-competent AAV or adenovirus (e.g., by a cytopathic effect bioassay).

The viral vector comprising the polynucleotide of the disclosure also include Retroviral vectors. Retroviruses are a class of integrative viruses which replicate using a virus-encoded reverse transcriptase, to replicate the viral RNA genome into double stranded DNA which is integrated into chromosomal DNA of the infected cells (e.g., target cells). Such vectors include those derived from murine leukemia viruses, especially Moloney (Gilboa, et al., 1988, Adv. Exp. Med. Biol. 241: 29) or Friend's FB29 strains (Int. Pat. Publ. No. WO1995/01447). Generally, a retroviral vector is deleted of all or part of the viral genes gag, pol and env and retains 5′ and 3′ LTRs and an encapsidation sequence. These elements may be modified to increase expression level or stability of the retroviral vector. Such modifications include the replacement of the retroviral encapsidation sequence by one of a retrotransposon such as VL30 (see, e.g., U.S. Pat. No. 5,747,323).

The polynucleotides encoding the polypeptide of the disclosure may be inserted downstream of the encapsidation sequence, such as in opposite direction relative to the retroviral genome. Retroviral particles are prepared in the presence of a helper virus or in an appropriate complementation (packaging) cell line which contains integrated into its genome the retroviral genes for which the retroviral vector is defective (e.g. gag/pol and env). Such cell lines are described in the prior art (Miller and Rosman, 1989, BioTechniques 7: 980; Danos and Mulligan, 1988, Proc. Natl. Acad. Sci. USA 85: 6460; Markowitz, et al., 1988, Virol. 167: 400). The product of the env gene is responsible for the binding of the viral particle to the viral receptors present on the surface of the target cell and, therefore determines the host range of the retroviral particle. Packaging cell line, such as the PA317 cells (ATCC CRL 9078) or 293EI6 (WO97/35996) containing an amphotropic envelope protein may therefore be used to allow infection of human and other species' target cells. The retroviral particles are recovered from the culture supernatant and may optionally be further purified according to standard techniques (e.g. chromatography, ultracentrifugation).

Self-Replicating RNA Molecules

Provided herein is a viral vector comprising any of the polynucleotides of the disclosure, wherein the vector is a self-replicating RNA molecule.

Self-replicating RNA may be derived from alphavirus. Alphaviruses may belong to the VEEV/EEEV group, or the SF group, or the SIN group. Non-limiting examples of SF group alphaviruses include Semliki Forest virus, O'Nyong-Nyong virus, Ross River virus, Middelburg virus, Chikungunya virus, Barmah Forest virus, Getah virus, Mayaro virus, Sagiyama virus, Bebaru virus, and Una virus. Non-limiting examples of SIN group alphaviruses include Sindbis virus, Girdwood S. A. virus, South African Arbovirus No. 86, Ockelbo virus, Aura virus, Babanki virus, Whataroa virus, and Kyzylagach virus. Non-limiting examples of VEEV/EEEV group alphaviruses include Eastern equine encephalitis virus (EEEV), Venezuelan equine encephalitis virus (VEEV), Everglades virus (EVEV), Mucambo virus (MUCV), Pixuna virus (PIXV), Middleburg virus (MIDV), Chikungunya virus (CHIKV), O'Nyong-Nyong virus (ONNV), Ross River virus (RRV), Barmah Forest virus (BF), Getah virus (GET), Sagiyama virus (SAGV), Bebaru virus (BEBV), Mayaro virus (MAYV), and Una virus (UNAV).

The self-replicating RNA molecules can be derived from alphavirus genomes, meaning that they have some of the structural characteristics of alphavirus genomes, or similar to them. The self-replicating RNA molecules can be derived from modified alphavirus genomes.

Self-replicating RNA molecules may be derived from Eastern equine encephalitis virus (EEEV), Venezuelan equine encephalitis virus (VEEV), Everglades virus (EVEV), Mucambo virus (MUCV), Semliki forest virus (SFV), Pixuna virus (PIXV), Middleburg virus (MIDV), Chikungunya virus (CHIKV), O'Nyong-Nyong virus (ONNV), Ross River virus (RRV), Barmah Forest virus (BF), Getah virus (GET), Sagiyama virus (SAGV), Bebaru virus (BEBV), Mayaro virus (MAYV), Una virus (UNAV), Sindbis virus (SINV), Aura virus (AURAV), Whataroa virus (WHAV), Babanki virus (BABV), Kyzylagach virus (KYZV), Western equine encephalitis virus (WEEV), Highland J virus (HJV), Fort Morgan virus (FMV), Ndumu (NDUV), and Buggy Creek virus. Virulent and avirulent alphavirus strains are both suitable. In some embodiments, the alphavirus RNA replicon is of a Sindbis virus (SIN), a Semliki Forest virus (SFV), a Ross River virus (RRV), a Venezuelan equine encephalitis virus (VEEV), or an Eastern equine encephalitis virus (EEEV).

In some embodiments, the alphavirus-derived self-replicating RNA molecule is a Venezuelan equine encephalitis virus (VEEV).

The self-replicating RNA molecules can contain RNA sequences from (or amino acid sequences encoded by) a wild-type New World or Old World alphavirus genome. Any of the self-replicating RNA molecules disclosed herein can contain RNA sequences “derived from” or “based on” wild type alphavirus genome sequences, meaning that they have at least 60% or at least 65% or at least 68% or at least 70% or at least 80% or at least 85% or at least 90% or at least 95% or at least 97% or at least 98% or at least 99% or 100% or 80-99% or 90-100% or 95-99% or 95-100% or 97-99% or 98-99% sequence identity with an RNA sequence (which can be a corresponding RNA sequence) from a wild type RNA alphavirus genome, which can be a New World or Old World alphavirus genome.

Self-replicating RNA molecules contain all of the genetic information required for directing their own amplification or self-replication within a permissive cell. To direct their own replication, self-replicating RNA molecules encode polymerase, replicase, or other proteins which may interact with viral or host cell-derived proteins, nucleic acids or ribonucleoproteins to catalyze the RNA amplification process; and contain cis-acting RNA sequences required for replication and transcription of the replicon-encoded RNA. Thus, RNA replication leads to the production of multiple daughter RNAs. These daughter RNAs, as well as collinear subgenomic transcripts, can be translated to provide in situ expression of a gene of interest, or can be transcribed to provide further transcripts with the same sense as the delivered RNA which are translated to provide in situ expression of the gene of interest. The overall results of this sequence of transcriptions is a huge amplification in the number of the introduced replicon RNAs and so the encoded gene of interest becomes a major polypeptide product of the cells.

There are two open reading frames (ORF's) in the genome of alphaviruses, non-structural (ns) and structural genes. The ns ORF encodes proteins (nsP1-nsP4) necessary for transcription and replication of viral RNA and are produced as a polyprotein and are the virus replication machinery. The structural ORF encodes three structural proteins: the core nucleocapsid protein C, and the envelope proteins P62 and El that associate as a heterodimer. The viral membrane-anchored surface glycoproteins are responsible for receptor recognition and entry into target cells through membrane fusion. The four ns protein genes are encoded by genes in the 5′ two-thirds of the genome, while the three structural proteins are translated from a subgenomic mRNA colinear with the 3′ one-third of the genome.

Self-replicating RNA molecules can be used as basis of introducing foreign sequences to host cells by replacing viral sequences encoding structural genes or inserting the foreign sequences 5′ or 3′ of the sequences encoding the structural genes. They can be engineered to replace the viral structural genes downstream of the replicase, which are under control of a subgenomic promoter, by genes of interest (GOI), e.g. the polynucleotide encoding for the polypeptide of the disclosure. Upon transfection, the replicase which is translated immediately, interacts with the 5′ and 3′ termini of the genomic RNA, and synthesizes complementary genomic RNA copies. Those act as templates for the synthesis of novel positive-stranded, capped, and poly-adenylated genomic copies, and subgenomic transcripts. Amplification eventually leads to very high RNA copy numbers of up to 2×10⁵ copies per cell. The result is a uniform and/or enhanced expression of a GOI (e.g. the polynucleotide encoding for the polypeptide of the disclosure) that can affect vaccine efficacy or therapeutic impact of a treatment. Vaccines based on self-replicating RNA molecules can therefore be dosed at very low levels due to the very high copies of RNA generated compared to conventional viral vector. One of the significant values of the compositions and methods disclosed herein is that vaccine efficacy can be increased in individuals that are in a chronic or acute state of immune activation.

The disclosure provides a self-replicating RNA molecule containing all of the genetic information required for directing its own amplification or self-replication within a permissive cell.

The disclosure also provides a self-replicating RNA molecule that can be used as the basis of introducing foreign sequences to host cells (e.g. the polypeptides of the disclosure) by replacing viral sequences encoding structural genes.

In some embodiments, the self-replicating RNA molecule comprises an RNA sequence encoding a protein or peptide; 5′ and 3′ alphavirus untranslated regions; RNA sequences encoding amino acid sequences derived from New World alphavirus VEEV nonstructural proteins nsP1, nsP2, nsP3 and nsP4; a sub-genomic promoter that is operably linked to and regulates translation of the RNA sequence encoding the protein; a 5′ cap and a 3′ poly-A tail; positive sense, single-stranded RNA; a DLP from Sindbis virus upstream of the non-structural protein 1(nsP1); a 2A ribosome skipping element; and a nsp1 nucleotide repeat downstream of the 5′-UTR and upstream of the DLP.

In some embodiments, the self-replicating RNA molecules may be at least 1 kb or at least 2 kb or at least 3 kb or at least 4 kb or at least 5 kb or at least 6 kb or at least 7 kb or at least 8 kb or at least 10 kb or at least 12 kb or at least 15 kb or at least 17 kb or at least 19 kb or at least 20 kb in size, or can be 100 bp-8 kb or 500 bp-8 kb or 500 bp-7 kb or 1-7 kb or 1-8 kb or 2-15 kb or 2-20 kb or 5-15 kb or 5-20 kb or 7-15 kb or 7-18 kb or 7-20 kb in size.

Any of the above-disclosed self-replicating RNA molecules can further include a coding sequence for an autoprotease peptide (e.g., autocatalytic self-cleaving peptide).

In some embodiments, the autoprotease peptide comprises, or consists of, a peptide sequence selected from the group consisting of porcine teschovirus-1 2A (P2A), a foot-and-mouth disease virus (FMDV) 2A (F2A), an Equine Rhinitis A Virus (ERAV) 2A (E2A), a Thosea asigna virus 2A (T2A), a cytoplasmic polyhedrosis virus 2A (BmCPV2A), a Flacherie Virus 2A (BmIFV2A), and a combination thereof. In some embodiments, the autoprotease peptide includes a peptide sequence of porcine teschovirus-1 2A (P2A).

Regulatory Elements

The polynucleotides encoding the polypeptides of the disclosure may be operably linked to one or more regulatory elements in the vector. The regulatory elements may comprise promoters, enhancers, polyadenylation signals, repressors and the like. As used herein, the term “operably linked” is to be taken in its broadest reasonable context and refers to a linkage of polynucleotide elements in a functional relationship. A polynucleotide is “operably linked” when it is placed into a functional relationship with another polynucleotide. For instance, a promoter is operably linked to a coding sequence if it affects the transcription of the coding sequence.

Some of the commonly used enhancer and promoter sequences in expression vectors and viral vectors are, for example, human cytomegalovirus (hCMV), vaccinia P7.5 early/late promoter, CAG, SV40, mouse CMV (mCMV), EF-1 and hPGK promoters. Due to its high potency and moderate size of ca. 0.8 kB, the hCMV promoter is one of the most commonly used of these promoters. The hPGK promoter is characterized by a small size (ca. 0.4 kB), but it is less potent than the hCMV promoter. On the other hand, the CAG promoter consisting of a cytomegalovirus early enhancer element, promoter, first exon and intron of chicken beta-actin gene, and splice acceptor of the rabbit beta-globin gene, can direct very potent gene expression that is comparable to the hCMV promoter, but its large size makes it less suitable in viral vectors where space constraints can be a significant concern, e.g., in adenoviral vectors (AdV), adeno-associated viral vectors (AAV) or lentiviral vectors (LVs).

Additional promoters that may be used are Aotine Herpesvirus 1 major immediate early promoter (AoHV-1 promoter) described in Int. Pat. Publ. No. WO2018/146205. The promoter may be operably coupled to a repressor operator sequence, to which a repressor protein can bind in order to repress expression of the promoter in the presence of the repressor protein. In certain embodiments, the repressor operator sequence is a TetO sequence or a CuO sequence (see e.g. U.S. Pat. No. 9,790,256).

In certain cases, it may be desirable to express at least two separate polypeptides from the same vector. In this case each polynucleotide may be operably linked to the same or different promoter and/or enhancer sequences, or well-known bicistronic expression systems for example by utilizing internal ribosome entry site (IRES) from encephalomyocarditis virus may be used. Alternatively, bidirectional synthetic promoters may be used, such as a hCMV-rhCMV promoter and other promoters described in Int. Pat. Publ. No. WO2017/220499. Polyadenylation signals may be derived from SV40 or bovine growth hormone (BGH).

The vectors comprising the polynucleotide encoding the polypeptide of the disclosure can further comprise any regulatory elements to establish conventional function(s) of the vector, including but not limited to replication and expression of the polypeptide of the disclosure encoded by the polynucleotide sequence of the vector. Regulatory elements include, but are not limited to, a promoter, an enhancer, a polyadenylation signal, translation stop codon, a ribosome binding element, a transcription terminator, selection markers, origin of replication, etc. A vector can comprise one or more expression cassettes. An “expression cassette” is part of a vector that directs the cellular machinery to make RNA and protein. An expression cassette typically comprises three components: a promoter sequence, an open reading frame, and a 3′-untranslated region (UTR) optionally comprising a polyadenylation signal. An open reading frame (ORF) is a reading frame that contains a coding sequence of a protein of interest (e.g., the polypeptides of the disclosure) from a start codon to a stop codon. Regulatory elements of the expression cassette can be operably linked to a polynucleotide sequence encoding the polypeptides of interest. Any components suitable for use in an expression cassette described herein can be used in any combination and in any order to prepare vectors of the application.

The vector can comprise a promoter sequence, preferably within an expression cassette, to control expression of the polypeptides of the disclosure. The term “promoter” is used in its conventional sense and refers to a nucleotide sequence that initiates the transcription of an operably linked nucleotide sequence. A promoter is located on the same strand near the nucleotide sequence it transcribes. Promoters can be a constitutive, inducible, or repressible. Promoters can be naturally occurring or synthetic. A promoter can be derived from sources including viral, bacterial, fungal, plants, insects, and animals. A promoter can be a homologous promoter (i.e., derived from the same genetic source as the vector) or a heterologous promoter (i.e., derived from a different vector or genetic source). Preferably, the promoter is located upstream of the polynucleotide encoding the polypeptides of the disclosure within an expression cassette. For example, in a self-replicating RNA, the promoter can be a subgenomic promoter for the alphavirus.

In a self-replicating RNA, the vector can further comprise additional polynucleotide sequences that stabilize the expressed transcript, enhance nuclear export of the RNA transcript, and/or improve transcriptional-translational coupling. Examples of such sequences include polyadenylation signals and enhancer sequences. A polyadenylation signal is typically located downstream of the coding sequence for a protein of interest (e.g., the polypeptides of the disclosure) within an expression cassette of the vector. Enhancer sequences are regulatory DNA sequences that, when bound by transcription factors, enhance the transcription of an associated gene. An enhancer sequence is preferably located upstream of the polynucleotide sequence encoding the polypeptides of the disclosure, but downstream of a promoter sequence within an expression cassette of the vector.

Any enhancer sequence known to those skilled in the art in view of the present disclosure can be used.

Any of the components or sequences of the vector of the disclosure can be functionally or operably linked to any other of the components or sequences. The components or sequences of the polypeptide fragments described herein can be operably linked for the expression of the at least one protein or peptide (or biotherapeutic) in a host cell or treated organism and/or for the ability of the replicon to self-replicate.

A promoter or UTR operably linked to a coding sequence is capable of effecting the transcription and expression of the coding sequence when the proper enzymes are present. The promoter need not be contiguous with the coding sequence, so long as it functions to direct the expression thereof Thus, an operable linkage between an RNA sequence encoding a protein or peptide and a regulatory sequence (for example, a promoter or UTR) is a functional link that allows for expression of the polynucleotide of interest. Operably linked can also refer to sequences such as the sequences encoding the RdRp (e.g. nsP4), nsP1-4, the UTRs, promoters, and other sequences encoding in the RNA replicon, are linked so that they enable transcription and translation of the biotherapeutic molecule and/or replication of the replicon. The UTRs can be operably linked by providing sequences and spacing necessary for recognition and translation by a ribosome of other encoded sequences.

Host Cells

The disclosure also provides a host cell comprising any of the above vectors of the disclosure.

In some embodiments, the host cell comprising any of the polynucleotides encoding the polypeptide fragments described herein is prokaryotic or eukaryotic host cell. In some embodiments, the host cell is PER.C6, PER.C6 TetO, a chicken embryo fibroblast (CEF), CHO, HEK293, HT-1080, HKB-11, CAP, HuH-7, or Age1 cell line.

In certain embodiments, the host cell comprising any of the polynucleotides encoding the TCRs described herein is a CD8+ T cell.

Compositions

The disclosure also provides compositions comprising any of the polynucleotides, any of the polypeptides, and any of the vectors disclosed herein. In some embodiments, the compositions may comprise a vector comprising any of the nucleotides disclosed herein.

Any of the compositions described above may comprise or may be formulated into a pharmaceutical composition comprising the composition and a pharmaceutically acceptable excipient. “Pharmaceutically acceptable” refers to the excipient that at the dosages and concentrations employed, will not cause unwanted or harmful effects in the subjects to which they are administered and include carrier, buffers, stabilizers or other materials well known to those skilled in the art. The precise nature of the carrier or other material may depend on the route of administration, e.g., intramuscular, subcutaneous, oral, intravenous, cutaneous, intramucosal (e.g., gut), intranasal or intraperitoneal routes. Liquid carriers such as water, petroleum, animal or vegetable oils, mineral oil or synthetic oil may be included. Physiological saline solution, dextrose or other saccharide solution or glycols such as ethylene glycol, propylene glycol or polyethylene glycol may be included. Exemplary formulation are the Adenovirus World Standard (Hoganson et al, 2002): 20 mM Tris pH 8, 25 mM NaCl, 2.5% glycerol; or 20 mM Tris, 2 mM MgCl2, 25 mM NaCl, sucrose 10% w/v, polysorbate-80 0.02% w/v; or 10-25 mM citrate buffer pH 5.9-6.2, 4-6% (w/w) hydroxypropyl-beta-cyclodextrin (HBCD), 70-100 mM NaCl, 0.018-0.035% (w/w) polysorbate-80, and optionally 0.3-0.45% (w/w) ethanol. Many other buffers can be used, and examples of suitable formulations for the storage and for pharmaceutical administration of purified pharmaceutical preparations are known.

The composition may comprise one or more adjuvants. Examples of such adjuvants include but are not limited to inorganic adjuvants (e.g. inorganic metal salts such as aluminium phosphate or aluminium hydroxide), organic adjuvants (e.g. saponins or squalene), oil-based adjuvants (e.g. Freund's complete adjuvant and Freund's incomplete adjuvant), liposomes, or biodegradable microspheres), virosomes, bacterial adjuvants (e.g. monophosphoryl lipid A, or muramyl peptides), synthetic adjuvants (e.g. non-ionic block copolymers, muramyl peptide analogues, or synthetic lipid A), or synthetic polynucleotides adjuvants (e.g polyarginine or polylysine). Other non-limiting examples of adjuvants include QS-21, Detox-PC, MPL-SE, MoGM-CSF, TiterMax-G, CRL-1005, GERBU, TERamide, PSC97B, Adjumer, PG-026, GSK-I, GcMAF, B-alethine, MPC-026, Adjuvax, CpG ODN, Betafectin, Alum, and MF59.

Other adjuvants that may be used include lectins, growth factors, cytokines, and lymphokines such as alpha-interferon, gamma interferon, platelet derived growth factor (PDGF), granulocyte-colony stimulating factor (GCSF), granulocyte macrophage colony stimulating factor (GMCSF), tumor necrosis factor (TNF), epidermal growth factor (EGF), IL-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12 or TLR agonists, and particulate adjuvants (e g immuno-stimulatory complexes (ISCOMS).

Methods of Treatment or Use

Provided herein are methods for treating a subject with the polypeptides, polynucleotides, vectors, or pharmaceutical compositions disclosed herein. The methods and uses provided herein comprise administering any of the polynucleotides, polypeptides, vectors, and compositions of the disclosure. The polynucleotides, polypeptides, vectors, compositions and administration regimens of the disclosure may be used to treat, prevent or reduce the risk of a clinical condition. The polynucleotides, polypeptides, vectors, compositions and administration regimens of the disclosure may be used to induce an immune response in a subject.

In certain embodiments the clinical condition is cancer. In certain embodiments, the cancer is characterized by expression of a neoantigen. In certain embodiments, the neoantigen is a polypeptide comprising an amino acid substitution, a frame shift mutation, a fusion, an in-frame deletion, or an insertion. In certain embodiments the neoantigen arises from overexpression of a polypeptide.

In certain embodiments, the clinical condition is characterized by expression of a CIC mutant. In some embodiments, the CIC mutant comprises an arginine to tryptophan amino acid substitution at a position corresponding to position 215 of SEQ ID NO: 102 (R215W).

In certain embodiments, the clinical condition is characterized by expression of a CTNNB1 mutant. In some embodiments, the CTNNB1 mutant comprises a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C). In some embodiments, the CTNNB1 mutant comprises a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F).

In certain embodiments, the clinical condition is characterized by expression of an ERBB2 mutant. In some embodiments, the ERBB2 mutant comprises a valine to isoleucine amino acid substitution at a position corresponding to position 842 of SEQ ID NO: 104 (V842I).

In certain embodiments, the clinical condition is characterized by expression of a KRAS mutant. In some embodiments, the KRAS mutant comprises a glycine to alanine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12A). In some embodiments, the KRAS mutant comprises a glycine to cysteine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12C). In some embodiments, the KRAS mutant comprises a glycine to valine amino acid substitution at a position corresponding to at position 12 of SEQ ID NO: 105 (G12V).

In certain embodiments, the clinical condition is characterized by expression of a PIK3CA mutant. In some embodiments, the PIK3CA mutant comprises a glutamic acid to lysine amino acid substitution at a position corresponding to position 453 of SEQ ID NO: 106 (E453K). In some embodiments, the PIK3CA mutant comprises a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D).

In certain embodiments, the clinical condition is characterized by expression of a PTEN mutant. In some embodiments, the PTEN mutant comprises an arginine to cysteine amino acid substitution at a position corresponding to position 173 of SEQ ID NO: 107 (R173C).

In certain embodiments, the clinical condition is characterized by expression of an SF3B1 mutant. In some embodiments, the SF3B1 mutant comprises an arginine to histidine amino acid substitution at a position corresponding to position 625 of SEQ ID NO: 108 (R625H).

In certain embodiments, the clinical condition is characterized by expression of a SOX17 mutant. In some embodiments, the SOX17 mutant comprises a serine to isoleucine amino acid substitution at a position corresponding to position 403 of SEQ ID NO: 109 (S403I).

In certain embodiments, the clinical condition is characterized by expression of a TP53 mutant. In some embodiments, the TP53 mutant comprises an arginine to leucine amino acid substitution at a position corresponding to position 110 of SEQ ID NO: 110 (R110L). In some embodiments, the TP53 mutant comprises a serine to phenylalanine amino acid substitution at a position corresponding to position 127 of SEQ ID NO: 110 (S127F). In some embodiments, the TP53 mutant comprises a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N). In some embodiments, the TP53 mutant comprises a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y). In some embodiments, the TP53 mutant comprises a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L). In some embodiments, the TP53 mutant comprises a histidine to leucine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193L). In some embodiments, the TP53 mutant comprises a histidine to tyrosine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193Y). In some embodiments, the TP53 mutant comprises a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C). In some embodiments, the TP53 mutant comprises a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M).

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of CIC mutant comprising an arginine to tryptophan amino acid substitution at a position corresponding to position 215 of SEQ ID NO: 102 (R215W) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 1, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 25, SEQ ID NO: 26, OR SEQ ID NO: 27, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of CIC mutant comprising an arginine to tryptophan amino acid substitution at a position corresponding to position 215 of SEQ ID NO: 102 (R215W) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 1, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 225, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of CTNNB1 mutant comprising a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 2, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, or SEQ ID NO: 80, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of CTNNB1 mutant comprising a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 2, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 29, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of CTNNB1 mutant comprising a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 2, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 226, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a CTNNB1 mutant comprising a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 3, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, or SEQ ID NO: 81, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a catenin beta 1 (CTNNB1) mutant comprising a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 3, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 32, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a catenin beta 1 (CTNNB1) mutant comprising a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 3, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 227, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a KRAS mutant comprising a glycine to alanine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12A) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 5, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 37, or SEQ ID NO: 38, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a KRAS mutant comprising a glycine to alanine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12A) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 5, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 228, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a KRAS mutant comprising a glycine to cysteine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12C) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 6, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 39, or SEQ ID NO: 40, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a KRAS mutant comprising a glycine to cysteine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12C) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 6, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 229, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a KRAS mutant comprising a glycine to valine amino acid substitution at a position corresponding to at position 12 of SEQ ID NO: 105 (G12V) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 7, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 41, or SEQ ID NO: 42, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a KRAS mutant comprising a glycine to valine amino acid substitution at a position corresponding to at position 12 of SEQ ID NO: 105 (G12V) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 7, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 230, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a PIK3CA mutant comprising a glutamic acid to lysine amino acid substitution at a position corresponding to position 453 of SEQ ID NO: 106 (E453K) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 8, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 43, or SEQ ID NO: 44, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a PIK3CA mutant comprising a glutamic acid to lysine amino acid substitution at a position corresponding to position 453 of SEQ ID NO: 106 (E453K) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 8, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 231, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a PIK3CA mutant comprising a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 9, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 45, SEQ ID NO: 46, or SEQ ID NO: 47, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a PIK3CA mutant comprising a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 9, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 45, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a PIK3CA mutant comprising a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 9, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 232, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a PTEN mutant comprising an arginine to cysteine amino acid substitution at a position corresponding to position 173 of SEQ ID NO: 107 (R173C) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 10, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 87, SEQ ID NO: 88, or SEQ ID NO: 89 or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a PTEN mutant comprising an arginine to cysteine amino acid substitution at a position corresponding to position 173 of SEQ ID NO: 107 (R173C) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 10, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 233 or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a SF3B1 mutant comprising an arginine to histidine amino acid substitution at a position corresponding to position 625 of SEQ ID NO: 108 (R625H) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 11, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 90, SEQ ID NO: 91, or SEQ ID NO: 92 or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a SF3B1 mutant comprising an arginine to histidine amino acid substitution at a position corresponding to position 625 of SEQ ID NO: 108 (R625H) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 11, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 234 or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a SOX10 mutant comprising a serine to isoleucine amino acid substitution at a position corresponding to position 403 of SEQ ID NO: 109 (S403I) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 12, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 93, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a SOX10 mutant comprising a serine to isoleucine amino acid substitution at a position corresponding to position 403 of SEQ ID NO: 109 (S403I) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 12, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 235, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 13, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, or SEQ ID NO: 94, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 13, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 59, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 13, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 244, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a histidine to leucine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193L) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 14, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 60, or SEQ ID NO: 61, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a histidine to leucine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193L) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 14, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 245, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a histidine to tyrosine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193Y) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 15, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 62, or SEQ ID NO: 63, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a histidine to tyrosine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193Y) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 15, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 236, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 16, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 64, or SEQ ID NO: 66, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 16, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 64, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 16, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 237, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 18, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 67, or SEQ ID NO: 68, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 18, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 68, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 18, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 239, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising an arginine to leucine amino acid substitution at a position corresponding to position 110 of SEQ ID NO: 110 (R110L) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 20, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 70, or SEQ ID NO: 71, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising an arginine to leucine amino acid substitution at a position corresponding to position 110 of SEQ ID NO: 110 (R110L) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 20, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 241, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a serine to phenylalanine amino acid substitution at a position corresponding to position 127 of SEQ ID NO: 110 (S127F) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 21, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 95, or SEQ ID NO: 96 or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. Also described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a serine to phenylalanine amino acid substitution at a position corresponding to position 127 of SEQ ID NO: 110 (S127F) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 21, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 242 or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 22, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 75, or SEQ ID NO: 76 or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 22, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 75, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 22, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 246, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 23, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 78, or SEQ ID NO: 79, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 23, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 78, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 23, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 243, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a ERBB2 mutant comprising a valine to isoleucine amino acid substitution at a position corresponding to position 842 of SEQ ID NO: 104 (V842I) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 24, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 84, SEQ ID NO: 85, or SEQ ID NO: 86, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition. In certain embodiments, described herein are methods of inducing an immune response or methods of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a ERBB2 mutant comprising a valine to isoleucine amino acid substitution at a position corresponding to position 842 of SEQ ID NO: 104 (V842I) in a subject, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 24, b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 247, or c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

In certain embodiments the methods of treatment comprise administering the polynucleotide in part a) prior to administering the polynucleotide in part b). In certain embodiments the methods of treatment comprise administering the polynucleotide in part b) prior to administering the polynucleotide in part a). In certain embodiments the methods of treatment comprise administering the polynucleotide in part a) concurrently with the polynucleotide in part b).

In certain embodiments, the time between administration of the polynucleotide in part a) and the polynucleotide in part b) is about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks, about 13 weeks, about 14 weeks, about 15 weeks, about 16 weeks, about 17 weeks, about 18 weeks, about 19 weeks, about 20 weeks, about 21 weeks, about 22 weeks, about 23 weeks, about 24 weeks, about 25 weeks, about 26 weeks, about 27 weeks, about 28 weeks, about 29 weeks, about 30 weeks, about 31 weeks, about 32 weeks, about 33 weeks, about 34 weeks, about 35 weeks, about 36 weeks, about 37 weeks, about 38 weeks, about 39 weeks, about 40 weeks, about 41 weeks, about 42 weeks, about 43 weeks, about 44 weeks, about 45 weeks, about 46 weeks, about 47 weeks, about 48 weeks, about 49 weeks, about 50 weeks, about 51 weeks, or about 52 weeks.

In certain embodiments the methods of treatment comprise administering a vector encoding the polynucleotide of part a) and a vector encoding the polynucleotide of part b). In some embodiments, the vectors are independently selected from an adenovirus vector, an alphaviral vector, a poxvirus vector, an adeno-associated virus vector, a retrovirus vector, and a self-replicating RNA molecule. In further embodiments, the adenovirus vector is selected from hAd5, hAd7, hAd11, hAd26, hAd34, hAd35, hAd48, hAd49, hAd50, GAd20, Gad19, GAd21, GAd25, GAd26, GAd27, GAd28, GAd29, GAd30, GAd31, ChAd3, ChAd4, ChAdS, ChAd6, ChAd7, ChAd8, ChAd9, ChAd10, ChAd11, ChAd16, ChAdI7, ChAd19, ChAd20, ChAd22, ChAd24, ChAd26, ChAd30, ChAd31, ChAd37, ChAd38, ChAd44, ChAd55, ChAd63, ChAd73, ChAd82, ChAd83, ChAd146, ChAd147, PanAd1, PanAd2, and PanAd3. In further embodiments, the poxvirus vector is selected from smallpox virus vector, vaccinia virus vector, cowpox virus vector, monkeypox virus vector, Copenhagen vaccinia virus (W) vector, New York Attenuated Vaccinia Virus (NYVAC) vector, and Modified Vaccinia Ankara (MVA) vector.

Also provided herein are methods of treating cancer in a subject comprising administering to the subject in need thereof a pharmaceutical composition comprising a TCR described herein. In certain embodiments, the TCR comprises an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a complementarity determining region 3 (CDR3) comprising an amino acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18, and the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18. An alpha chain CDR3 corresponds to a beta chain CDR3 if they appear in the same row in Table 14, Table 15, Table 16, Table 17 or Table 18.

In further embodiments, the TCR comprises an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, a CDR2 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, and a CDR3 comprising an amino acid sequence provided in Table 14, Table 15, Table 16, Table 17, or Table 18, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, a CDR2 comprising an amino acid sequence provided in Table 19, Table 20, Table 21, Table 22, or Table 23, and a CDR3 comprising a corresponding amino acid sequence provided in Table 14, Table 15, Table 16, Table 17 or Table 18. An alpha chain CDR1 or CDR2 corresponds to a beta chain CDR1 or CDR2 if they appear in the same row in Table 19, Table 20, Table 21, Table 22, or Table 23. An alpha chain CDR3 corresponds to a beta chain CDR3 if they appear in the same row in Table 14, Table 15, Table 16, Table 17, or Table 18. An alpha and beta chain CDR1 and CDR2 provided in Table 19, Table 20, Table 21, Table 22, or Table 23 correspond to and alpha and beta chain CDR3 provided in the same row in Table 14, Table 15, Table 16, Table 17 or Table 18.

Also provided herein are methods of inducing an immune response in a subject comprising administering to the subject in need thereof a pharmaceutical composition comprising a TCR described herein.

Described herein are methods of inducing an immune response or treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a PIK3CA mutant comprising a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D) in a subject comprising administering to the subject in need thereof a TCR described herein. In certain embodiments, the TCR comprises an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 14, and (b) the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 14. In certain embodiments, the TCR comprises an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 19, a CDR2 comprising an amino acid sequence provided in Table 19, and a CDR3 comprising an amino acid sequence provided in Table 14, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 19, a CDR2 comprising an amino acid sequence provided in Table 19, and a CDR3 comprising a corresponding amino acid sequence provided in Table 14.

Described herein are methods of inducing an immune response or treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y) in a subject comprising administering to the subject in need thereof a TCR described herein. In certain embodiments, the TCR comprises an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 15, and (b) the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 15. In certain embodiments, the TCR comprises an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 20, a CDR2 comprising an amino acid sequence provided in Table 20, and a CDR3 comprising an amino acid sequence provided in Table 15, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 20, a CDR2 comprising an amino acid sequence provided in Table 20, and a CDR3 comprising a corresponding amino acid sequence provided in Table 15.

Described herein are methods of inducing an immune response or treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L) in a subject comprising administering to the subject in need thereof a TCR described herein. In certain embodiments, the TCR comprises an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 16, and (b) the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 16. In certain embodiments, the TCR comprises an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 21, a CDR2 comprising an amino acid sequence provided in Table 21, and a CDR3 comprising an amino acid sequence provided in Table 16, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 21, a CDR2 comprising an amino acid sequence provided in Table 21, and a CDR3 comprising a corresponding amino acid sequence provided in Table 16.

Described herein are methods of inducing an immune response or treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a CTNNB1 mutant comprising a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F) in a subject comprising administering to the subject in need thereof a TCR described herein. In certain embodiments, the TCR comprises an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 17, and (b) the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 17. In certain embodiments, the TCR comprises an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 22, a CDR2 comprising an amino acid sequence provided in Table 22, and a CDR3 comprising an amino acid sequence provided in Table 17, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 22, a CDR2 comprising an amino acid sequence provided in Table 22, and a CDR3 comprising a corresponding amino acid sequence provided in Table 17.

Described herein are methods of inducing an immune response or treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a TP53 mutant comprising a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C) in a subject comprising administering to the subject in need thereof a TCR described herein. In certain embodiments, the TCR comprises an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 18, and (b) the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 18. In certain embodiments, the TCR comprises an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 23, a CDR2 comprising an amino acid sequence provided in Table 23, and a CDR3 comprising an amino acid sequence provided in Table 18, and (b) the beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 23, a CDR2 comprising an amino acid sequence provided in Table 23, and a CDR3 comprising a corresponding amino acid sequence provided in Table 18.

Kits/Articles of Manufacture

For use in the methods or uses described herein, kits and articles of manufacture are also described. Such kits include a package or container that is compartmentalized to receive one or more dosages of the pharmaceutical compositions disclosed herein. Suitable containers include, for example, bottles. In one embodiment, the containers are formed from a variety of materials such as glass or plastic.

Described herein are kits of parts comprising a pair of polypeptide fragments selected from the group consisting of: (a) SEQ ID NO: 2 and SEQ ID NO: 29; (b) SEQ ID NO: 3 and SEQ ID NO: 32; (c) SEQ ID NO: 9 and SEQ ID NO: 45; (d) SEQ ID NO: 13 and SEQ ID NO: 59; (e) SEQ ID NO: 16 and SEQ ID NO: 64; (0 SEQ ID NO: 18 and SEQ ID NO 68; (g) SEQ ID NO: 22 and SEQ ID NO: 75; and (h) SEQ ID NO: 23 and SEQ ID NO: 78.

Described herein are kits of parts comprising a pair of polypeptide fragments selected from the group consisting of: (a) SEQ ID NO: 9 and SEQ ID NO: 45; (b) SEQ ID NO: 13 and SEQ ID NO: 59; and (c) SEQ ID NO: 18 and SEQ ID NO: 68.

The articles of manufacture provided herein contain packaging materials. Packaging materials for use in packaging pharmaceutical products include, e.g., U.S. Pat. Nos. 5,323,907, 5,052,558 and 5,033,252. Examples of pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubes, bags, containers, bottles, and any packaging material suitable for a selected formulation and intended mode of administration and treatment.

A kit typically includes labels listing contents and/or instructions for use, and package inserts with instructions for use. A set of instructions will also typically be included.

In one embodiment, a label is on or associated with the container. In one embodiment, a label is on a container when letters, numbers or other characters forming the label are attached, molded or etched into the container itself; a label is associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert.

In one embodiment, a label is used to indicate that the contents are to be used for a specific therapeutic application. The label also indicates directions for use of the contents, such as in the methods described herein.

In certain embodiments, the pharmaceutical compositions are presented in a pack or dispenser device which contains one or more unit dosage forms containing a compound provided herein. The pack, for example, contains metal or plastic foil, such as a blister pack. In one embodiment, the pack or dispenser device is accompanied by instructions for administration. In one embodiment, the pack or dispenser is also accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, is the labeling approved by the U.S. Food and Drug Administration for prescription drugs, or the approved product insert. In one embodiment, compositions containing a compound provided herein formulated in a compatible pharmaceutical carrier are also prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

Methods of Generating CD8+ T-Cells

Described herein are methods for generating CD8+ T-cells that are positive for an HLA-A*02:01-restricted polypeptide fragment and a cognate native polypeptide fragment. In certain embodiments, said methods comprise exposing CD8+ T-cells to the HLA-A*02:01-restricted polypeptide fragment and cognate native polypeptide fragment selected from the group consisting of: (a) SEQ ID NO: 2 or a sequence having 90% identity to SEQ ID NO: 2 and SEQ ID NO: 29 or a sequence having 90% identity to SEQ ID NO: 29; (b) SEQ ID NO: 3 or a sequence having 90% identity to SEQ ID NO: 3 and SEQ ID NO: 32 or a sequence having 90% identity to SEQ ID NO: 32; (c) SEQ ID NO: 9 or a sequence having 90% identity to SEQ ID NO: 9 and SEQ ID NO: 45 or a sequence having 90% identity to SEQ ID NO: 45; (d) SEQ ID NO: 13 or a sequence having 90% identity to SEQ ID NO: 13 and SEQ ID NO: 59 or a sequence having 90% identity to SEQ ID NO: 59; (e) SEQ ID NO: 16 or a sequence having 90% identity to SEQ ID NO: 16 and SEQ ID NO: 64 or a sequence having 90% identity to SEQ ID NO: 64; (f) SEQ ID NO: 18 or a sequence having 90% identity to SEQ ID NO: 18 and SEQ ID NO 68 or a sequence having 90% identity to SEQ ID NO: 68; (g) SEQ ID NO: 22 or a sequence having 90% identity to SEQ ID NO: 22 and SEQ ID NO: 75 or a sequence having 90% identity to SEQ ID NO: 75; and (h) SEQ ID NO: 23 or a sequence having 90% identity to SEQ ID NO: 23 and SEQ ID NO: 78 or a sequence having 90% identity to SEQ ID NO: 78, and selecting CD8+ T cells that are positive to both the HLA-A*02:01-restricted polypeptide fragment and a cognate neoantigen polypeptide fragment.

Described herein are methods for generating CD8+ T-cells that are positive for an HLA-A*02:01-restricted polypeptide fragment and a cognate native polypeptide fragment. In certain embodiments, said methods comprise exposing CD8+ T-cells to the HLA-A*02:01-restricted polypeptide fragment and cognate native polypeptide fragment selected from the group consisting of: (a) SEQ ID NO: 9 or a sequence having 90% identity to SEQ ID NO: 9 and SEQ ID NO: 45 or a sequence having 90% identity to SEQ ID NO: 45; (b) SEQ ID NO: 13 or a sequence having 90% identity to SEQ ID NO: 13 and SEQ ID NO: 59 or a sequence having 90% identity to SEQ ID NO: 59; and (c) SEQ ID NO: 18 or a sequence having 90% identity to SEQ ID NO: 18 and SEQ ID NO 68 or a sequence having 90% identity to SEQ ID NO: 68; and selecting CD8+ T cells that are positive to both the HLA-A*02:01-restricted polypeptide fragment and a cognate neoantigen polypeptide fragment.

EXAMPLES

These examples are provided for illustrative purposes only and not to limit the scope of the claims provided herein.

Example 1

Peptide Mimic Design

Selection of Cancer Driver Mutations

A list of cancer driver mutations compiled from 9176 cancer patients was examined as described in Marty R, et al. Cell. 2017; 171(6):1272-1283, and driver mutations were prioritized based on their recurrence. HLA binding predictions using the NetMHCpan4.0 suite of prediction algorithms were performed as described in Jurtz V., et al., J Immunol. 2017; 199(9):3360-3368, and antigens were prioritized on the basis of their predicted binding affinities to HLA-A*02:01. The HLA-A*02:01 allele was chosen for mimic design due to its high allele frequency (˜40%) in North American and European populations, and because there is abundant experimental binding data of different antigens bound to HLA-A*02:01. Finally, mimic antigens were designed as follows.

Prioritization of Mutations and Mimic Design

Driver mutations in cancer are expected to be present in most of the tumor cells (i.e. they have high clonality). The mutant proteins can be processed into 9- or 10-mer neo-antigen epitopes that contain the mutation and are presented by the class I MHC complex of these tumor cells. Neo-epitopes that have very weak binding affinity to MHC molecules are not presented since they are unable to stabilize the peptide-MHC complex, while epitopes with intermediate or strong binding affinity to the MHC I protein could be presented on the surface of tumor cells. Furthermore, the stability of the peptide-MHC complex can determine the number and duration of the complex on the cell surface. As a result, the immunogenicity of a given antigen is related to its binding affinity to MHC I proteins.

Cancer driver mutations arise early in the process of malignant transformation of cells into cancers, and they are subject to surveillance by the immune system. Hence, cells with mutations that can result in highly immunogenic antigens are likely to be recognized and killed by the immune system. As a consequence, early cancer driver mutations are expected to be weakly immunogenic and to have moderate binding to the host MHC proteins. On the other hand, due to their high clonality, driver mutations are an attractive target for therapies that redirect the immune system to specifically recognize and attack cancer cells. Thus, an approach has been developed based on molecular mimicry, wherein synthetic mimic peptide antigens are designed with enhanced MHC binding affinity relative to the driver mutant antigens, but with sufficient sequence similarity that they elicit an immune response that can also recognize and attack the mutant antigens presented on cancer cells.

Methods

9-mer and 10-mer epitopes containing a cancer mutation were identified, and mimics were designed against those candidate epitopes that exhibited intermediate (weak) predicted binding affinities to the HLA-A*02:01 allele. To analyze the predicted binding affinities, NetMHCpan4.0 was used, which ranks epitopes according to their predicted affinity to a given HLA allele, with lower ranks being indicative of a higher binding affinity. The ranks of epitopes are allowed to vary across a range from 0 to 100. Most natural peptide epitopes were identified as predicted to bind at a rank <2. Motivated by this feature of the algorithm, the predicted binding was classified as intermediate if the rank of the binding affinity was >0.5 and predicted binding affinity was <4, and strong if the rank was <0.5. Mimics were designed by allowing amino acid substitutions at amino acid positions 2 and 9 (P2 and P9) of the binding 9mer epitope. To generate rules for amino acid substitutions, the set of known 9mer epitope binders to HLA-A*02:01 was obtained from the IEDB database (Vita R, et al. Nucleic Acids Res. 2018 Oct 24), and the frequencies of occurrence of different amino acids at positions P2 and P9 estimated.

The Shannon entropy was estimated for each amino acid substitution at P2 and P9 as a measure of amino acid conservation. To generate mimic peptides, amino acids at P2 and P9 were ranked by degree of conservation and substitutions by replacing the wild type amino acid by other amino acid residues according to their degree of conservation. Amino acid conservation at P2 in the order L>M>I and at P9 in the order V>L>I was used. Finally, mimics for antigens with the cancer driver mutation present at either of P2 or P9 (the anchor positions on a 9mer epitope) were designed by altering amino acids at the anchor position which did not contain the mutation, thus retaining the cancer driver mutation.

The mimic peptides were then again examined for their predicted binding affinity with netMHCpan4.0 and ranked accordingly. A list of 90 peptides (23 cancer driver mutation+66 mimic epitopes, excluding controls, for an average of 3 mimics/mutant) was created by ranking cancer mutant 9- and 10-mer epitopes and their corresponding mimic pairs according to the ratio of their predicted affinities. Mutant and mimic sequences are provided in Table 1. Mutant peptide substitutions relative to wild-type 9- or 10-mer epitopes are identified with bold font and mimic peptide amino acid substitutions relative to the mutant peptides are identified with underlining.

TABLE 1
					SEQ
					ID
Name	Type	Gene	Variant	Sequence	NO
M010	mutant	CIC	CIC.R215W	MIFSKRHWA	1
M011	mimic	CIC	CIC_R215W15	MMFSKRHWI	25
M012	mimic	CIC	CIC_R215W6	MLFSKRHWV	26
M013	mimic	CIC	CIC_R215W7	MMFSKRHWV	27
M020	mutant	CTNNB1	CTNNB1.S33C	YLDCGIHSG	2
M021	mimic	CTNNB1	CTNNB1_S33C10	YMDCGIHSL	28
M022	mimic	CTNNB1	CTNNB1_S33C5	YLDCGIHSV	29
M023	mimic	CTNNB1	CTNNB1_S33C6	YMDCGIHSV	30
M030	mutant	CTNNB1	CTNNB1.S37F	YLDSGIHFG	3
M031	mimic	CTNNB1	CTNNB1_S37F14	YMDSGIHFI	31
M032	mimic	CTNNB1	CTNNB1_S37F5	YLDSGIHFV	32
M033	mimic	CTNNB1	CTNNB1_S37F6	YMDSGIHFV	33
M034	mimic	CTNNB1	CTNNB1.S33C9	YLDCGIHSL	80
M035	mimic	CTNNB1	CTNNB1.S37F10	YMDSGIHFL	81
M040	control	Control	N/A	YLSTDVGFA	4
M041	mimic	Control	N/A	YLSTDVGFV	34
M042	mimic	Control	N/A	YMSTDVGFV	35
M043	mimic	Control	N/A	YLSTDVGFL	36
M044	mimic	Control	N/A	YMSTDVGFL	82
M045	mimic	Control	N/A	YLSTDVGFI	83
M050	mutant	KRAS	KRAS.G12A	LVVVGAAGV	5
M051	mimic	KRAS	KRAS_G12A2	LLVVGAAGV	37
M052	mimic	KRAS	KRAS_G12A3	LMVVGAAGV	38
M060	mutant	KRAS	KRAS.G12C	LVVVGACGV	6
M061	mimic	KRAS	KRAS_G12C2	LLVVGACGV	39
M062	mimic	KRAS	KRAS_G12C3	LMVVGACGV	40
M070	mutant	KRAS	KRAS.G12V	LVVVGAVGV	7
M071	mimic	KRAS	KRAS_G12V2	LLVVGAVGV	41
M072	mimic	KRAS	KRAS_G12V3	LMVVGAVGV	42
M080	mutant	PIK3CA	PIK3CA.E453K	GLKDLLNPI	8
M081	mimic	PIK3CA	PIK3CA_E453K5	GLKDLLNPV	43
M082	mimic	PIK3CA	PIK3CA_E453K6	GMKDLLNPV	44
M090	mutant	PIK3CA	PIK3CA.G118D	ILNREIDFA	9
M091	mimic	PIK3CA	PIK3CA_G118D5	ILNREIDFV	45
M092	mimic	PIK3CA	PIK3CA_G118D6	IMNREIDFV	46
M093	mimic	PIK3CA	PIK3CA_G118D9	ILNREIDFL	47
M100	mutant	PTEN	PTEN.R173C	CYVYYYSYLL	10
M101	mimic	PTEN	PTEN R173C2	CYLYYYSYLL	48
M102	mimic	PTEN	PTEN R173C6	CYLYYYSYLV	49
M103	mimic	PTEN	PTEN R173C7	CYMYYYSYLV	50
M104	mimic	PTEN	PTEN.R173C3	CYMYYYSYLL	87
M105	mimic	PTEN	PTEN.R173C10	CYLYYYSYLI	88
M106	mimic	PTEN	PTEN.R173C11	CYMYYYSYLI	89
M110	mutant	SF3B1	SF3B1.R625H	NMDEYVHNT	11
M111	mimic	SF3B1	SF3B1_R625H5	NMDEYVHNV	51
M112	mimic	SF3B1	SF3B1_R625H6	NLDEYVHNV	52
M113	mimic	SF3B1	SF3B1_R625H9	NMDEYVHNL	53
M114	mimic	SF3B1	SF3B1.R625H10	NLDEYVHNL	90
M115	mimic	SF3B1	SF3B1.R625H13	NMDEYVHNI	91
M116	mimic	SF3B1	SF3B1.R625H14	NLDEYVHNI	92
M120	mutant	SOX17	SOX17.S4031	VVSDAISAV	12
M121	mimic	SOX17	SOX17_S40312	VLSDAISAV	54
M122	mimic	SOX17	SOX17_S40313	VMSDAISAV	55
M123	mimic	SOX17	SOX17_S40316	VLSDAISAL	56
M124	mimic	SOX17	SOX17.S40317	VMSDAISAL	93
M130	mutant	TP53	TP53.C141Y	KTYPVQLWV	13
M131	mimic	TP53	TP53_C141Y2	KLYPVQLWV	57
M132	mimic	TP53	TP53_C141Y3	KMYPVQLWV	58
M133	mimic	TP53	TP53_C141Y8	KMYPVQLWL	59
M134	mimic	TP53	TP53.C141Y12	KLYPVQLWI	94
M140	mutant	TP53	TP53.H193L	GLAPPQLLI	14
M141	mimic	TP53	TP53_H193L5	GLAPPQLLV	60
M142	mimic	TP53	TP53_H193L6	GMAPPQLLV	61
M150	mutant	TP53	TP53.H193Y	GLAPPQYLI	15
M151	mimic	TP53	TP53_H193Y5	GLAPPQYLV	62
M152	mimic	TP53	TP53_H193Y6	GMAPPQYLV	63
M160	mutant	TP53	TP53.K132N	ALNNMFCQL	16
M161	mimic	TP53	TP53_K132N5	ALNNMFCQV	64
M162	mimic	TP53	TP53_K132N6	AMNNMFCQV	66
M170	mutant	TP53	TP53.K132N	NMFCQLAKT	17
M171	mimic	TP53	TP53_K132N6	N LFCQLAKV	65
M180	mutant	TP53	TP53.P152L	QLWVDSTPL	18
M181	mimic	TP53	TP53_P152L3	QLWVDSTPI	67
M182	mimic	TP53	TP53_P152L4	QLWVDSTPV	68
M190	mutant	TP53	TP53.P250L	RLILTIITL	19
M191	mimic	TP53	TP53_P250L4	RLILTIITV	69
M200	mutant	TP53	TP53.R110L	YQGSYGFLL	20
M201	mimic	TP53	TP53_R110L3	YQGSYGFLI	70
M202	mimic	TP53	TP53_R110L4	YQGSYGFLV	71
M210	mutant	TP53	TP53.S127F	SVTCTYFPA	21
M211	mimic	TP53	TP53_S127F11	SMTCTYFPL	72
M212	mimic	TP53	TP53_S127F6	SLTCTYFPV	73
M213	mimic	TP53	TP53_S127F7	SMTCTYFPV	74
M214	mimic	TP53	TP53_S127F8	SLTCTYFPL	95
M215	mimic	TP53	TP53_S127F12	SMTCTYFPI	96
M220	mutant	TP53	TP53.V272M	LLGRNSFEM	22
M221	mimic	TP53	TP53_V272M2	LLGRNSFEL	75
M222	mimic	TP53	TP53_V272M3	LLGRNSFEI	76
M223	Wild	TP53	TP53_V272M4	LLGRNSFEV	77
	type
M230	mutant	TP53	TP53.Y220C	VVPCEPPEV	23
M231	mimic	TP53	TP53_Y220C2	VLPCEPPEV	78
M232	mimic	TP53	TP53_Y220C3	VMPCEPPEV	79
M240	mutant	ERBB2	ERBB2.V842I	RLIHRDLAA	24
M241	mimic	ERBB2	ERBB2_V842110	RMIHRDLAL	84
M242	mimic	ERBB2	ERBB2_V842I5	RLIHRDLAV	85
M243	mimic	ERBB2	ERBB2_V84216	RMIHRDLAV	86
M250	wild	CMV	CMV.pp65	NLVPMVATV	97
	type
	control
M251	mimic	CMV	CMV.pp651	NMVPMVATV	98
M252	mimic	CMV	CMV.pp652	NLVPMVATL	99
M253	mimic	CMV	CMV.pp653	NMVPMVATL	100

The wild-type amino acid sequences for each of the genes identified in Table 1 are provided in Table 2.

TABLE 2
			Protein
		SEQ	Data-
		ID	base
Gene	Amino Acid Sequence	NO	No.
CIC	MYSAHRPLMPASSAASRGLGMFVWTNVEPRSVAVFPWHSLVPFLA	102	Q96RK0
	PSQPDPSVQPSEAQQPASHPVASNQSKEPAESAAVAHERPPGGTG
	SADPERPPGATCPESPGPGPPHPLGVVESGKGPPPTTEEEASGPP
	GEPRLDSETESDHDDAFLSIMSPEIQLPLPPGKRRTQSLSALPKE
	RDSSSEKDGRSPNKREKDHIRRPMNAFMIFSKRHRALVHQRHPNQ
	DNRTVSKILGEWWYALGPKEKQKYHDLAFQVKEAHFKAHPDWKWC
	NKDRKKSSSEAKPTSLGLAGGHKETRERSMSETGTAAAPGVSSEL
	LSVAAQTLLSSDTKAPGSSSCGAERLHTVGGPGSARPRAFSHSGV
	HSLDGGEVDSQALQELTQMVSGPASYSGPKPSTQYGAPGPFAAPG
	EGGALAATGRPPLLPTRASRSQRAASEDMTSDEERMVICEEEGDD
	DVIADDGFGTTDIDLKCKERVTDSESGDSSGEDPEGNKGFGRKVF
	SPVIRSSFTHCRPPLDPEPPGPPDPPVAFGKGYGSAPSSSASSPA
	SSSASAATSFSLGSGTFKAQESGQGSTAGPLRPPPPGAGGPATPS
	KATRFLPMDPATFRRKRPESVGGLEPPGPSVIAAPPSGGGNILQT
	LVLPPNKEEQEGGGARVPSAPAPSLAYGAPAAPLSRPAATMVTNV
	VRPVSSTPVPIASKPFPTSGRAEASPNDTAGARTEMGTGSRVPGG
	SPLGVSLVYSDKKSAAATSPAPHLVAGPLLGTVGKAPATVTNLLV
	GTPGYGAPAPPAVQFIAQGAPGGGTTAGSGAGAGSGPNGPVPLGI
	LQPGALGKAGGITQVQYILPTLPQQLQVAPAPAPAPGTKAAAPSG
	PAPTTSIRFTLPPGTSTNGKVLAATAPTPGIPILQSVPSAPPPKA
	QSVSPVQAPPPGGSAQLLPGKVLVPLAAPSMSVRGGGAGQPLPLV
	SPPFSVPVQNGAQPPSKIIQLTPVPVSTPSGLVPPLSPATLPGPT
	SQPQKVLLPSSTRITYVQSAGGHALPLGTSPASSQAGTVTSYGPT
	SSVALGFTSLGPSGPAFVQPLLSAGQAPLLAPGQVGVSPVPSPQL
	PPACAAPGGPVITAFYSGSPAPTSSAPLAQPSQAPPSLVYTVATS
	TTPPAATILPKGPPAPATATPAPTSPFPSATAGSMTYSLVAPKAQ
	RPSPKAPQKVKAAIASIPVGSFEAGASGRPGPAPRQPLEPGPVRE
	PTAPESELEGQPTPPAPPPLPETWTPTARSSPPLPPPAEERTSAK
	GPETMASKFPSSSSDWRVPGQGLENRGEPPTPPSPAPAPAVAPGG
	SSESSSGRAAGDTPERKEAAGTGKKVKVRPPPLKKTFDSVDNRVL
	SEVDFEERFAELPEFRPEEVLPSPTLQSLATSPRAILGSYRKKRK
	NSTDLDSAPEDPTSPKRKMRRRSSCSSEPNTPKSAKCEGDIFTFD
	RTGTEAEDVLGELEYDKVPYSSLRRTLDQRRALVMQLFQDHGFFP
	SAQATAAFQARYADIFPSKVCLQLKIREVRQKIMQAATPTEQPPG
	AEAPLPVPPPTGTAAAPAPTPSPAGGPDPTSPSSDSGTAQAAPPL
	PPPPESGPGQPGWEGAPQPSPPPPGPSTAATGR
CTNNB1	MATQADLMELDMAMEPDRKAAVSHWQQQSYLDSGIHSGATTTAPS	103	P35222
	LSGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTR
	AQRVRAAMFPETLDEGMQIPSTQFDAAHPTNVQRLAEPSQMLKHA
	VVNLINYQDDAELATRAIPELTKLLNDEDQVVVNKAAVMVHQLSK
	KEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLSHHR
	EGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAK
	MAVRLAGGLQKMVALLNKTNVKFLAITTDCLQILAYGNQESKLII
	LASGGPQALVNIMRTYTYEKLLWTTSRVLKVLSVCSSNKPAIVEA
	GGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGMEGLLGTL
	VQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRT
	VLRAGDREDITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVV
	KLLHPPSHWPLIKATVGLIRNLALCPANHAPLREQGAIPRLVQLL
	VRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGALHILARDVH
	NRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEA
	IEAEGATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLS
	VELTSSLFRTEPMAWNETADLGLDIGAQGEPLGYRQDDPSYRSFH
	SGGYGQDALGMDPMMEHEMGGHHPGADYPVDGLPDLGHAQDLMDG
	LPPGDSNQLAWFDTDL
ERBB2	MELAALCRWGLLLALLPPGAASTQVCTGTDMKLRLPASPETHLDM	104	P04626
	LRHLYQGCQVVQGNLELTYLPTNASLSFLQDIQEVQGYVLIAHNQ
	VRQVPLQRLRIVRGTQLFEDNYALAVLDNGDPLNNF1PVTGASPG
	GLRELQLRSLTEILKGGVLIQRNPQLCYQDTILWKDIFHKNNQLA
	LTLIDTNRSRACHPCSPMCKGSRCWGESSEDCQSLTRTVCAGGCA
	RCKGPLPTDCCHEQCAAGCTGPKHSDCLACLHFNHSGICELHCPA
	LVTYNTDTFESMPNPEGRYTFGASCVTACPYNYLSTDVGSCTLVC
	PLHNQEVTAEDGTQRCEKCSKPCARVCYGLGMEHLREVRAVTSAN
	IQEFAGCKKIFGSLAFLPESFDGDPASNTAPLQPEQLQVFETLEE
	ITGYLYISAWPDSLPDLSVFQNLQVIRGRILHNGAYSLTLQGLGI
	SWLGLRSLRELGSGLALIHHNTHLCFVHTVPWDQLFRNPHQALLH
	TANRPEDECVGEGLACHQLCARGHCWGPGPTQCVNCSQFLRGQEC
	VEECRVLQGLPREYVNARHCLPCHPECQPQNGSVTCFGPEADQCV
	ACAHYKDPPFCVARCPSGVKPDLSYMPIWKFPDEEGACQPCPINC
	THSCVDLDDKGCPAEQRASPLTSIISAVVGILLVVVLGVVFGILI
	KRRQQKIRKYTMRRLLQETELVEPLTPSGAMPNQAQMRILKETEL
	RKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANK
	EILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHV
	RENRGRLGSQDLLNWCMQIAKGMSYLEDVRLVHRDLAARNVLVKS
	PNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFT
	HQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPP
	ICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQ
	NEDLGPASPLDSTFYRSLLEDDDMGDLVDAEEYLVPQQGFFCPDP
	APGAGGMVHHRHRSSSTRSGGGDLTLGLEPSEEEAPRSPLAPSEG
	AGSDVFDGDLGMGAAKGLQSLPTHDPSPLQRYSEDPTVPLPSETD
	GYVAPLTCSPQPEYVNQPDVRPQPPSPREGPLPAARPAGATLERP
	KTLSPGKNGVVKDVFAFGGAVENPEYLTPQGGAAPQPHPPPAFSP
	AFDNLYYWDQDPPERGAPPSTFKGTPTAENPEYLGLDVPV
KRAS	MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV	105	P01116
	IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSF
	EDIHHYREQIKRVKDSEDVPMVLVGNKCDLPSRTVDTKQAQDLAR
	SYGIPFIETSAKTRQRVEDAFYTLVREIRQYRLKKISKEEKTPGC
	VKIKKCIIM
PIK3CA	MPPRPSSGELWGIHLMPPRILVECLLPNGMIVTLECLREATLITI	106	P42336
	KHELFKEARKYPLHQLLQDESSYIFVSVTQEAEREEFFDETRRLC
	DLRLFQPFLKVIEPVGNREEKILNREIGFAIGMPVCEFDMVKDPE
	VQDFRRNILNVCKEAVDLRDLNSPHSRAMYVYPPNVESSPELPKH
	IYNKLDKGQIIVVIWVIVSPNNDKQKYTLKINHDCVPEQVIAEAI
	RKKTRSMLLSSEQLKLCVLEYQGKYILKVCGCDEYFLEKYPLSQY
	KYIRSCIMLGRMPNLMLMAKESLYSQLPMDCFTMPSYSRRISTAT
	PYMNGETSTKSLWVINSALRIKILCATYVNVN1RDIDKIYVRTGI
	YHGGEPLCDNVNTQRVPCSNPRWNEWLNYDIYIPDLPRAARLCLS
	ICSVKGRKGAKEEHCPLAWGNINLFDYTDTLVSGKMALNLWPVPH
	GLEDLLNPIGVTGSNPNKETPCLELEFDWFSSVVKFPDMSVIEEH
	ANWSVSREAGFSYSHAGLSNRLARDNELRENDKEQLKAISTRDPL
	SEITEQEKDFLWSHRHYCVTIPEILPKLLLSVKWNSRDEVAQMYC
	LVKDWPPIKPEQAMELLDCNYPDPMVRGFAVRCLEKYLTDDKLSQ
	YLIQLVQVLKYEQYLDNLLVRFLLKKALTNQRIGHFFFWHLKSEM
	ENKTVSQRFGLLLESYCRACGMYLKHLNRQVEAMEKLINLTDILK
	QEKKDETQKVQMKFLVEQMRRPDFMDALQGFLSPLNPAHQLGNLR
	LEECRIMSSAKRPLWLNWENPDIMSELLFQNNEI1FKNGDDLRQD
	MLTLQIIRIMENIWQNQGLDLRMLPYGCLSIGDCVGLIEVVRNSH
	TIMQIQCKGGLKGALQFNSHTLHQWLKDKNKGEIYDAAIDLFTRS
	CAGYCVATFILGIGDRHNSNIMVKDDGQLFHIDFGHFLDHKKKKF
	GYKRERVPFVLTQDFLIVISKGAQECTKTREFERFQEMCYKAYLA
	IRQHANLFINLFSMMLGSGMPELQSFDDIAYIRKTLALDKTEQEA
	LEYFMKQMNDAHHGGWTTKMDWIFHTIKQHALN
PTEN	MTAIIKEIVSRNKRRYQEDGFDLDLTYIYPNIIAMGFPAERLEGV	107	P60484
	YRNNIDDVVRFLDSKHKNHYKIYNLCAERHYDTAKFNCRVAQYPF
	EDHNPPQLELIKPFCEDLDQWLSEDDNHVAAIHCKAGKGRTGVMI
	CAYLLHRGKFLKAQEALDFYGEVRTRDKKGVTIPSQRRYVYYYSY
	LLKNHLDYRPVALLFHKMMFETIPMFSGGTCNPQFVVCQLKVKIY
	SSNSGPTRREDKFMYFEFPQPLPVCGDIKVEFFHKQNKMLKKDKM
	FHFWVNTFFIPGPEETSEKVENGSLCDQEIDSICSIERADNDKEY
	LVLTLTKNDLDKANKDKANRYFSPNFKVKLYFTKTVEEPSNPEAS
	SSTSVTPDVSDNEPDHYRYSDTTDSDPENEPFDEDQHTQITKV
SF3B1	MAKIAKTHEDIEAQIREIQGKKAALDEAQGVGLDSTGYYDQEIYG	108	075333
	GSDSRFAGYVTSIAATELEDDDDDYSSSTSLLGQKKPGYHAPVAL
	LNDIPQSTEQYDPFAEHRPPKIADREDEYKKHRRTMIISPERLDP
	FADGGKTPDPKMNARTYMDVMREQHLTKEEREIRQQLAEKAKAGE
	LKVVNGAAASQPPSKRKRRWDQTADQTPGATPKKLSSWDQAETPG
	HTPSLRWDETPGRAKGSETPGATPGSKIWDPTPSHTPAGAATPGR
	GDTPGHATPGHGGATSSARKNRWDETPKTERDTPGHGSGWAETPR
	TDRGGDSIGETPTPGASKRKSRWDETPASQMGGSTPVLTPGKTPI
	GTPAMNMATPTPGHIMSMTPEQLQAWRWEREIDERNRPLSDEELD
	AMFPEGYKVLPPPAGYVPIRTPARKLTATPTPLGGMTGFHMQTED
	RTMKSVNDQPSGNLPFLKPDDIQYFDKLLVDVDESTLSPEEQKER
	KIMKLLLKIKNGTPPMRKAALRQITDKAREFGAGPLFNQILPLLM
	SPTLEDQERHLLVKVIDRILYKLDDLVRPYVHKILVVIEPLLIDE
	DYYARVEGREIISNLAKAAGLATMISTMRPDIDNMDEYVRNTTAR
	AFAVVASALGIPSLLPFLKAVCKSKKSWQARHTGIKIVQQIAILM
	GCAILPHLRSLVEIIEHGLVDEQQKVRTISALAIAALAEAATPYG
	IESFDSVLKPLWKGIRQHRGKGLAAFLKAIGYLIPLMDAEYANYY
	TREVMLILIREFQSPDEEMKKIVLKVVKQCCGTDGVEANYIKTEI
	LPPFFKHFWQHRMALDRRNYRQLVDTTVELANKVGAAEIISRIVD
	DLKDEAEQYRKMVMETIEKIMGNLGAADIDHKLEEQLIDGILYAF
	QEQTTEDSVMLNGFGTVVNALGKRVKPYLPQICGTVLWRLNNKSA
	KVRQQAADLISRTAVVMKTCQEEKLMGHLGVVLYEYLGEEYPEVL
	GSILGALKAIVNVIGMHKMTPPIKDLLPRLTPILKNRHEKVQENC
	IDLVGRIADRGAEYVSAREWMRICFELLELLKAHKKAIRRATVNT
	FGYIAKAIGPHDVLATLLNNLKVQERQNRVCTTVAIAIVAETCSP
	FTVLPALMNEYRVPELNVQNGVLKSLSFLFEYIGEMGKDYIYAVT
	PLLEDALMDRDLVHRQTASAVVQHMSLGVYGFGCEDSLNHLLNYV
	WPNVFETSPHVIQAVMGALEGLRVAIGPCRMLQYCLQGLFHPARK
	VRDVYWKIYNSIYIGSQDALIAHYPRIYNDDKNTYIRYELDYIL
SOX17	MSSPDAGYASDDQSQTQSALPAVMAGLGPCPWAESLSPIGDMKVK	109	Q9H6I2
	GEAPANSGAPAGAAGRAKGESRIRRPMNAFMVWAKDERKRLAQQN
	PDLHNAELSKMLGKSWKALTLAEKRPFVEEAERLRVQHMQDHPNY
	KYRPRRRKQVKRLKRVEGGFLHGLAEPQAAALGPEGGRVAMDGLG
	LQFPEQGFPAGPPLLPPHMGGHYRDCQSLGAPPLDGYPLPTPDTS
	PLDGVDPDPAFFAAPMPGDCPAAGTYSYAQVSDYAGPPEPPAGPM
	HPRLGPEPAGPSIPGLLAPPSALHVYYGAMGSPGAGGGRGFQMQP
	QHQHQHQHQHHPPGPGQPSPPPEALPCRDGTDPSQPAELLGEVDR
	TEFEQYLHFVCKPEMGLPYQGHDSGVNLPDSHGAISSVVSDASSA
	VYYCNYPDV
TP53	MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML	110	P04637
	SPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPS
	WPLSSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFC
	QLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHE
	RCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPPEV
	GSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSF
	EVRVCACPGRDRRTEEENLRKKGEPHHELPPGSTKRALPNNTSSS
	PQPKKKPLDGEYFTLQIRGRERFEMFRELNEALELKDAQAGKEPG
	GSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD
CMV	MESRGRRCPEMISVLGPISGHVLKAVFSRGDTPVLPHETRLLQTG	111	P06725
	IHVRVSQPSLILVSQYTPDSTPCHRGDNQLQVQHTYFTGSEVENV
	SVNVHNPTGRSICPSQEPMSIYVYALPLKMLNIPSINVHHYPSAA
	ERKHRHLPVADAVIHASGKQMWQARLTVSGLAWTRQQNQWKEPDV
	YYTSAFVFPTKDVALRHVVCAHELVCSMENTRATKMQVIGDQYVK
	VYLESFCEDVPSGKLFMHVTLGSDVEEDLTMTRNPQPFMRPHERN
	GFTVLCPKNMIIKPGKISHIMLDVAFTSHEHFGLLCPKSIPGLSI
	SGNLLMNGQQIFLEVQAIRETVELRQYDPVAALFFFDIDLLLQRG
	PQYSEHPTFTSQYRIQGKLEYRHTWDRHDEGAAQGDDDVWTSGSD
	SDEELVTTERKTPRVTGGGAMAGASTSAGRKRKSASSATACTSGV
	MTRGRLKAESTVAPEEDTDEDSDNEIHNPAVFTWPPWQAGILARN
	LVPMVATVQGQNLKYQEFFWDANDIYRIFAELEGVWQPAAQPKRR
	RHRQDALPGPCIASTPKKHRG

Example 2

Screen for Best Binders to HLA-A*02:01

The 9- or 10-mer peptides from Table 1 were synthesized and MHC binding assays were performed to experimentally evaluate their binding to HLA-A*02:01 as follows.

UV-Mediated Peptide Exchange

Overview

HLA-bound peptides are critical for the stability of the HLA complex. A conditional HLA class I complex is stabilized by an UV-labile peptide (p*). Through UV irradiation this peptide can be cleaved in the HLA-bound state. Because the obtained peptide fragments no longer meet the strict length requirement for high-affinity HLA class I binding, these fragments dissociate from the HLA class I complex and the complex disintegrates. Under the conditions in which peptide cleavage is performed (neutral pH, on melting ice), the resulting peptide-free HLA complex is stable, and when cleavage is performed in the presence of another HLA class I peptide, this reaction results in net exchange of the cleaved peptide, yielding an HLA class I complex with an epitope of choice. The peptide exchange efficiency can be analyzed using an HLA class I ELISA. The combined technologies allow the identification of ligands for an HLA molecule of interest which are potentially immunogenic.

Exchange control peptide Pos is a high affinity binder to the relevant HLA class I allele while exchange control peptide Neg is a non-binder. The UV control represents UV-irradiation of conditional HLA class I complex in the absence of a rescue peptide. The binding of exchange control peptide Neg and all experimental peptides were evaluated relative to that of exchange control peptide Pos. The absorption of the latter peptide is put to 100%. This procedure results in a range of different percentages depending on the affinities of the different experimental peptides for the HLA allele that is used. An arbitrary cut off value was chosen as a positive cut off for binders.

Assay Procedure

All reagents were brought to 0° C. by putting them on melting ice. The concentrated p*HLA*02:01 (1.5 mg/mL) class I solution was kept in the dark to ensure stability. All vials were centrifuged at 3000 g for 1 minute before use.

Preparation of the peptides of choice: 4594 sterile phosphate buffered saline pH 7.4 (PBS) and 5 μL of peptide (10 mg/mL) were pipetted in 1.4 mL Micronic tubes (Micronic #MP32022). Then, using a multichannel pipetting device, 10 μL of diluted peptide was added to a 384-well PP microtiter plate.

P*HLA class I solution: The concentrated p*HLA*02:01 (1.5 mg/mL) class I solution was diluted in an amber safe lock tube to 50 μg/mL in PBS and kept on melting ice in the dark.

Preparation of change controls: A stock solution of change control positive (abbreviated “pos”) for p*HLA*02:01 (NLVPMVATV) (SEQ ID NO: 97) was prepared at 10 mg/mL in 100% dimethyl sulfoxide (DMSO). A stock solution of change control negative (abbreviated “neg”) for p*HLA*02:01 (IVTDFSVIK) (SEQ ID NO: 101) was prepared at 10 mg/mL in 100% DMSO. The positive and negative stock solutions were diluted to 100 μM using 5 μL peptide and 495 μL PBS. Three tubes were labeled for each mixture: ‘Pos,’ ‘Neg,’ and ‘UV.’ The following reagents were added per tube:

	TABLE 3
	Reagent	Pos	Neg	UV
	PBS	—	—	12.5 μL
	Change control Pos (100 μM)	12.5 μL	—	—
	Change control Neg (100 μM)	—	12.5 μL	—
	Diluted p*HLA class I solution	12.5 μL	12.5 μL	12.5 μL

20 μL of the controls were mixed and transferred to the 384-well plate.

UV-induced peptide exchange: 10 μL of the diluted p*HLA class I solution was pipetted into the 384-well PP microtiter plate using a multichannel pipetting device into each well, and the solution was mixed thoroughly using the multichannel pipetting device. The plate was sealed and centrifuged at 3300 g for 2 minutes at 4° C. The seal was removed, and the plate was placed on ice and under the ultraviolet (UV) lamp for 30 minutes with the UV lamp at a distance of 2-5 cm from the sample. The plate was sealed and incubated for 30 minutes at 37° C. The plate was centrifuged at 3300 g for 5 minutes at 4° C. Two UV-induced peptide exchanges were performed (“exchange I” and “exchange II”).

Screening: The outcome of the UV-mediated HLA peptide exchange was evaluated by HLA class I ELISA.

Enzyme Immunoassay for the Determination of the Presence of Intact HLA Class I Complexes

Overview

The HLA class I ELISA is an enzyme immunoassay based on the detection of beta2-microglobulin (B2M) of (peptide-stabilized) HLA class I complexes. To this end streptavidin is bound onto polystyrene microtiter wells. After washing and blocking, HLA complex present in exchange reaction mixtures or ELISA controls is captured by the streptavidin on the microtiter plate via its biotinylated heavy chain. Non-bound material is removed by washing. Subsequently, horseradish peroxidase (HRP)-conjugated antibody to human B2M is added. This antibody binds only to an intact HLA complex present in the microtiter well because unsuccessful peptide exchange results in disintegration of the original UV-sensitive HLA complex upon UV illumination. In the latter case B2M is removed during the washing step. After removal of non-bound HRP conjugate by washing, a substrate solution is added to the wells. A colored product is formed in proportion to the amount of intact HLA complex present in the samples. After the reaction has been terminated by the addition of a stop solution, absorbance is measured in a microtiter plate reader. The absorbance is normalized to the absorbance of an exchange control peptide (represents 100%). Also, suboptimal HLA binding of peptides with a moderate to low affinity for HLA class I molecules can be detected by this ELISA technique

Assay Procedure

Before use, all reagents were brought to room temperature (18-25° C.) with the exception of anti-human beta2-microglobulin-HRP conjugate and a screen control (2.7 μM HLA complex), which were kept on melting ice to ensure stability. All vials were centrifuged before use (1 minute 3000 g).

Coating wells of two NUNC MaxiSorp™ 96-well ELISA plates: only the contents of one coating buffer capsule were dissolved in 100 mL of distilled water (0.05 M carbonate-bicarbonate buffer, pH 9.6 at 25° C.). 46 μL of Streptavidin stock solution were added to 23 mL coating buffer. 100 μL of the Streptavidin stock solution were added to all wells. Each microtiter plate was covered with an adhesive seal and incubated overnight at room temperature (18-25° C.).

Dilution buffer (Sanquin): The exchange reaction mixtures and controls were diluted in working-strength Dilution buffer.

Washing procedure (Sanquin): Fresh Washing buffer was prepared. Supernatants were discarded from wells and the wells were filled with Washing buffer (300 μL per well) and tipped out. This was repeated three times.

Blocking procedure: 300 μL of working-strength Dilution buffer was added to all wells. The microtiter plate(s) were covered with adhesive seal or lid and incubated for 30 minutes at room temperature (18-25° C.).

Preparation of ELISA HLA controls: From the Screen control three HLA controls were generated by serial dilution in Dilution buffer. The controls were prepared fresh and kept on melting ice until usage. Specifically, 4 tubes were labeled, one tube for each dilution: ‘1:500’, ‘H’, ‘M’ and ‘L’. 1.5 mL of working-strength Dilution buffer was pipetted into the tube ‘1:500’ and 500 μL into the other tubes. 3 μL of the Screen control was transferred into the first tube labeled ‘1:500’, mixed well, and 500 μL of this dilution was transferred into the second tube labeled ‘H’. The serial dilution was repeated twice by adding 500 μL of the previous tube of diluted control to the 500 μL of working-strength Dilution buffer.

Dilution of exchange reaction mixtures: To evaluate the outcome of UV-mediated HLA peptide exchange, a small aliquot of the exchange reaction mixture was diluted in working-strength Dilution buffer (the proper dilution factor was p*HLA lot-dependent). The exchange reaction mixture was diluted in working-strength Dilution buffer.

Incubation step (controls and exchange reaction mixtures): Dilution buffer was tipped out from the wells. 100 μL of working-strength Dilution buffer was pipetted into the blank wells and 100 μL of the HLA controls was pipetted into in the appropriate wells. 100 μL of the prepared exchange reaction mixture dilutions was transferred into the appropriate wells. The plates were covered with adhesive seal and incubated for 1 hour at 37° C.

Wash step: supernatant was discarded from the wells and the microtiter plates were washed as described in ‘Washing procedure’ above.

Incubation step (HRP-conjugated antibody): Per microtiter plate, 11 μL of concentrated HRP-conjugated antibody was added to 11 mL of working-strength Dilution buffer just before use. 100 μL of diluted HRP-conjugated antibody was added to all wells. The plates were covered with adhesive seal and incubate for 1 hour at 37° C.

Wash step: The supernatant was discarded from the wells and the microtiter plates were washed as described in ‘Washing procedure’ above.

Incubation step (enzymatic color development): Approximately 10 minutes before use, the substrate solution was prepared as follows per microtiter plate: 9.57 mL of distilled water; 1.1 mL of Substrate buffer stock solution; 220 μL of ABTS stock solution; and 110 μL of Hydrogen peroxide stock solution.

The substrate solution was at room temperature (18-25° C.). 100 μL of substrate solution was added to all wells and the wells were incubated for 8 minutes at room temperature (18-25° C.) in the dark on a plate shaker at 400-500 rpm.

Stop enzymatic reaction: 50 μL of Stop buffer (Sanquin) was added to all wells.

Plate read-out: plates were read at 414 nm in an ELISA reader within 30 minutes.

Results

The results from the two UV-mediated HLA peptide exchanges (abbreviated ‘Exch I’ and ‘Exch II’) are provided in Table 4. “SD” stands for standard deviation.

TABLE 4
SEQ		Exch I	Exch II	Average
ID NO	Sequence	(%)	(%)	%	SD
Pos	NLVPMVATV	100.0	100.0	100.0	0.0000
(97)
Neg	IVTDFSVIK	7.0	6.9	6.9	0.0081
(101)
UV	No	7.9	7.6	7.8	0.2293
	peptide
1	MIFSKRHWA	6.2	6.1	6.2	0.0842
2	YLDCGIHSG	34.8	33.0	33.9	1.2804
3	YLDSGIHFG	53.7	53.2	53.4	0.4031
5	LVVVGAAGV	6.6	6.2	6.4	0.2337
6	LVVVGACGV	6.5	7.5	7.0	0.7515
7	LVVVGAVGV	4.3	4.6	4.5	0.2227
8	GLKDLLNPI	73.8	84.9	79.4	7.7994
9	ILNREIDFA	53.1	47.8	50.4	3.7270
10	CYVYYYSYL	38.2	38.5	38.4	0.2562
	L
11	NMDEYVHNT	9.2	7.3	8.3	1.3162
12	VVSDAISAV	88.5	83.8	86.2	3.2840
13	KTYPVQLWV	66.6	71.2	68.9	3.2688
14	GLAPPQLLI	9.2	7.2	8.2	1.4007
15	GLAPPQYLI	13.5	13.1	13.3	0.2875
16	ALNNMFCQL	39.6	43.1	41.3	2.4847
18	QLWVDSTPL	84.3	88.1	86.2	2.6915
20	YQGSYGFLL	14.3	9.6	11.9	3.3633
21	SVTCTYFPA	7.7	9.4	8.6	1.1462
22	LLGRNSFEM	58.8	42.9	50.9	11.2708
23	VVPCEPPEV	29.4	37.5	33.5	5.7534
24	RLIHRDLAA	6.8	7.7	7.2	0.6221
25	MMFSKRHWI	40.4	45.3	42.8	3.4162
26	MLFSKRHWV	74.8	73.8	74.3	0.6809
27	MMFSKRHWV	76.2	72.6	74.4	2.5217
28	YMDCGIHSL	107.9	103.3	105.6	3.2934
29	YLDCGIHSV	100.4	103.5	101.9	2.2217
30	YMDCGIHSV	100.8	115.1	108.0	10.0855
31	YMDSGIHFI	97.6	115.8	106.7	12.8512
32	YLDSGIHFV	99.3	108.7	104.0	6.6311
33	YMDSGIHFV	104.1	95.5	99.8	6.0739
37	LLVVGAAGV	34.1	32.5	33.3	1.1431
38	LMVVGAAGV	51.0	51.5	51.3	0.3763
39	LLVVGACGV	36.4	33.1	34.8	2.3173
40	LMVVGACGV	47.1	50.4	48.7	2.3509
41	LLVVGAVGV	31.8	40.1	35.9	5.8591
42	LMVVGAVGV	39.0	33.3	36.1	4.0013
43	GLKDLLNPV	73.5	76.3	74.9	1.9449
44	GMKDLLNPV	74.7	80.4	77.5	4.0472
45	ILNREIDFV	62.1	78.7	70.4	11.7708
46	IMNREIDFV	68.6	64.9	66.7	2.5838
47	ILNREIDFL	51.1	61.3	56.2	7.1945
48	CYLYYYSYL	16.1	20.6	18.3	3.1585
	L
49	CYLYYYSYL	10.4	10.9	10.7	0.3435
	V
51	NMDEYVHNV	86.6	80.6	83.6	4.2367
52	NLDEYVHNV	88.1	88.4	88.2	0.1818
53	NMDEYVHNL	62.0	54.3	58.1	5.4566
54	VLSDAISAV	89.3	90.9	90.1	1.1771
55	VMSDAISAV	96.9	87.9	92.4	6.3778
56	VLSDAISAL	91.9	82.6	87.3	6.5884
57	KLYPVQLWV	84.1	79.8	81.9	3.0527
58	KMYPVQLWV	72.7	81.0	76.9	5.8253
59	KMYPVQLWL	122.1	102.8	112.5	13.6215
60	GLAPPQLLV	40.3	44.1	42.2	2.6661
61	GMAPPQLLV	13.9	13.9	13.9	0.0076
62	GLAPPQYLV	48.8	47.6	48.2	0.8672
63	GMAPPQYLV	22.5	20.3	21.4	1.5723
64	ALNNMFCQV	69.7	68.8	69.3	0.6662
66	AMNNMFCQV	69.4	70.8	70.1	1.0420
67	QLWVDSTPI	62.0	73.8	67.9	8.3729
68	QLWVDSTPV	95.6	95.7	95.7	0.0913
70	YQGSYGFLI	7.1	10.8	8.9	2.6418
71	YQGSYGFLV	53.6	45.6	49.6	5.6164
72	SMTCTYFPL	56.4	50.6	53.5	4.0810
73	SLTCTYFPV	93.7	97.2	95.5	2.4364
74	SMTCTYFPV	83.4	93.3	88.4	7.0148
75	LLGRNSFEL	101.2	95.7	98.4	3.9238
76	LLGRNSFEI	75.3	58.0	66.7	12.2339
78	VLPCEPPEV	66.3	73.7	70.0	5.1976
79	VMPCEPPEV	62.9	58.9	60.9	2.8611
80	YLDCGIHSL	102.8	110.9	106.8	5.7387
81	YMDSGIHFL	101.5	107.8	104.7	4.4311
84	RMIHRDLAL	51.5	55.8	53.7	3.0145
85	RLIHRDLAV	69.9	77.3	73.6	5.2146
86	RMIHRDLAV	56.7	58.5	57.6	1.2275
87	CYMYYYSYL	8.3	8.7	8.5	0.2510
	L
88	CYLYYYSYL	27.8	18.9	23.3	6.3290
	I
89	CYMYYYSYL	4.9	4.8	4.9	0.0760
	I
90	NLDEYVHNL	57.2	53.0	55.1	2.9456
91	NMDEYVHNI	73.6	63.1	68.4	7.4102
92	NLDEYVHNI	71.1	69.0	70.1	1.4997
93	VMSDAISAL	75.3	79.5	77.4	2.9691
94	KLYPVQLWI	81.3	66.4	73.9	10.5149
95	SLTCTYFPL	69.9	71.6	70.8	1.2464
96	SMTCTYFPI	32.2	22.7	27.5	6.7081
97	NLVPMVATV	99.4	89.1	94.3	7.2778
98	NMVPMVATV	77.6	66.2	71.9	8.0381
99	NLVPMVATL	26.8	20.4	23.6	4.5505
100	NMVPMVATL	26.4	22.0	24.2	3.0889

Conclusions

Peptides having SEQ ID NOs: 28-32, 59, 80, and 81 showed binding of more than 100% as compared to the positive peptide control (average relative binding to HLA-A*02:01 in the range of 102-113%). Peptides having SEQ ID NOs: 33, 54, 55, 68, 73, 75, and 97 showed an average binding in the range of 90-100%. The average binding of peptides having SEQ ID NOs: 12, 18, 51, 52, 56, 57, and 74 was in the range of 80-88%. Peptides having SEQ ID NOs: 8, 26, 27, 43-45, 58, 66, 78, 85, 92-95, and 98 demonstrated an average binding in the range of 70-79%. The average binding of peptides having SEQ ID NOs: 13, 46, 64, 67, 76, 79, and 91 was in the range of 61-69%. Peptides having SEQ ID NOs: 3, 9, 22, 38, 47, 53, 71, 72, 84, 86, and 90 demonstrated an average relative binding to HLAA*02:01 in the range of 50-58%. Peptides having SEQ ID NOs: 16, 25, 40, 60, and 62 showed an average binding in the range of 41-49%. The average binding of peptides having SEQ ID NOs: 2, 10, 23, 37, 39, 41, and 42 was in the range of 33-38%. Peptides having SEQ ID NOs: 48, 63, 88, 96, 99, and 100 showed an average binding in the range of 18-28%. The average binding of all other peptides was found to be below 15%.

The experimental values of binding were used to prioritize 8 mutant-mimic pairs for further experimental validation (Table 5) based on two criteria—i) the mutant epitopes have moderate binding to HLA-A*0201 (>10% in comparison to CMV pp65 antigen) and ii) the ratio of mimic binding to mutant binding was as high as possible (a ratio of >1).

TABLE 5
			No.
			patients
	SEQ		out of			SEQ
	ID		9176	Mutant		ID	Mimic
Mutant	NO	Mutation	patients1	Binding	Mimic	NO	Binding
YLDCGIHSG	2	CTNNB1.S33C	13	34.8	YLDCGIHSV	29	101.9
YLDSGIHFG	3	CTNNB1.S37F	21	53.7	YLDSGIHFV	32	104.0
ILNREIDFA	9	PIK3CA.G118D	19	50.4	ILNREIDFV	45	70.4
KTYPVQLWV	13	TP53.C141Y	14	68.9	KMYPVQLWL	59	112.5
ALNNMFCQL	16	TP53.K132N	19	41.3	ALNNMFCQV	64	69.3
QLWVDSTPL	18	TP53.P152L	10	86.2	QLWVDSTPV	68	95.7
LLGRNSFEM	22	TP53.V272M	19	50.9	LLGRNSFEL	75	98.4
VVPCEPPEV	23	TP53.Y220C	62	33.5	VLPCEPPEV	78	70.0
1Marty R, et al. Cell. 2017;171(6):1272-1283

Example 3

TCRs that Recognize Mutant Peptides are Cross-Reactive to Mimic Peptides Determination of Frequency of Double Positive T Cells for Mimic and Mutant Tetramers from HLA-02:01⁺Primary Peripheral Blood Mononuclear Cells (PBMCs)

Protocol

Preparation of PBMCs: Vials of HLA-02:01⁺PBMCs frozen from donors (Hemacare) were removed from LN₂ storage and rapidly thawed in a 37° C. water bath. The cells were transferred to a 50 mL conical tube containing 40mL warm media (RPMI 1640 medium+10% fetal bovine serum (FBS)+ 1% Penicillin streptomycin solution), spun at 1300 rpm for 5 minutes at room temperature. The cell pellet was resuspended in 2 mL EASYSEP™ buffer and cells were counted using trypan blue live dead marker using a haemocytometer.

Enrichment of CD8+ T cells: To enrich the CD8+ T cells from the PBMCs, EASYSEP™ Human CD8+ T Cell Isolation Kit was used as per the manufacturer's instructions. Post enrichment, the cell pellet was resuspended in Dulbecco's phosphate-buffered saline (DPBS) and cells were counted as above. To determine the viability, LIVE/DEAD™ Fixable Violet Dead Cell Stain Kit was used at 0.5 μL/1×10⁶/100 μL cell suspension and incubated at room temperature for 20 minutes. At the end of incubation period, FACS buffer (DPBS +2% FBS) was added and the cells were spun at 1300 rpm for 5 minutes at room temperature. The cell pellet was resuspended in FACS buffer, trypan blue was added, and the cell count was determined using a haemocytometer. The cell density was maintained at 1×10⁶/50 μL FACS buffer. 3 μL Fc block was added per 1×10⁶ cells and incubated for 10 minutes at room temperature in the dark. At the end of the incubation, mimic tetramer and the corresponding mutant tetramers were added at 3 μL tetramer/1×10⁶ cells. The sequences of the peptides used for tetramer synthesis is provided in Table 6.

TABLE 6
		Tetramer			Tetramer
		Conjugate			Conjugate
Mutated	SEQ	for	Mimic	SEQ	for
peptide	ID	mutant	peptide	ID	mimic
sequence	NO	sequence	sequence	NO	seq.
ILNREIDFA	9	PE	ILNREIDFV	45	APC
KTYPVQLWV	13	APC	KMYPVQLWL	59	PE
QLWVDSTPL	18	APC	QLWVDSTPV	68	PE
YLDCGIHSG	2	APC	YLDCGIHSV	29	PE
YLDSGIHFG	3	APC	YLDSGIHFV	32	PE
ALNNMFCQL	16	PE	ALNNMFCQV	64	APC
LLGRNSFEM	22	PE	LLGRNSFEL	75	APC
VVPCEPPEV	23	PE	VLPCEPPEV	78	APC

For frequency determination, 2×10⁶ cells were used for test samples and 1×10⁶cells were used for control samples where negative tetramers were added in place of the mimic or mutant tetramers. A sample where no tetramers were added was kept as unstained control. The samples were incubated at room temperature for 30 minutes in dark. At the end of incubation period, CD8 antibody was added at 2 μL/1×10⁶cells and the samples were incubated for another 30 minutes at room temperature in dark. At the end of incubation, FACS buffer was added to the samples and the samples were spun at 1300 rpm for 5 minutes. The pellet was resuspended in 5 mL FACS buffer and the cells were spun at 1300 rpm for 5 minutes. The pellet was resuspended in 200 μL FACS buffer and events were acquired using the Novocyte flow cytometer.

Gating Strategy and Data Analysis

The cells were acquired on the Novocyte flow cytometer and gated on forward scatter height (FSC-H) versus side scatter height (SSC-H). The cells high on FSC-H versus SSC-H were gated as lymphocytes. From the lymphocytes, live cells were gated by selecting the pacific blue negative cells on an FSC-H versus Pacific Blue-H plot. From the live cells, single cells were gated on FSC-H versus forward scatter area (FSC-A). CD8+ T cells were gated from the singlets as Alexa Fluor 700 positive cells. For gating phycoerythrin (PE) conjugated tetramer positive cells, the CD8⁺ cells were gated on Alexa Fluor 700-H versus PE-H and the double positive cells were considered as CD8+ tetramer+. For gating allophycocyanin (APC) conjugated tetramer positive cells, the CD8⁺ cells were gated on Alexa Fluor 700-H versus APC-H and the double positive cells were considered as CD8+ tetramer+. All gates were set using the negative tetramers to eliminate non-specific binding. The dual positive cells that were mimic and mutant tetramer positive cells were gated by plotting PE-H versus APC-H within the CD8⁺ cells. The percentage of double positive cells for APC and PE is the frequency of mimic and mutant positive tetramer cells displayed in the gate.

Sorting of Double Positive Cells for Mimic and Mutant Tetramers Specific to Peptide Sequences from HLA-02:01⁺ PBMCs

Protocol

PBMCs were prepared and CD8+ T cells were enriched following the protocol described above. At the end of the 10 minute room temperature incubation in 3 μL Fc block, mimic tetramer and the corresponding mutant tetramers were added.

For sorting, a minimum of 5×10⁶ cells were used for test samples. 1×10⁶ cells were used for control samples where negative tetramers were added in place of the mimic or mutant tetramers and a sample where no tetramers were added was kept as unstained control. 3 μL tetramer/1×10⁶ cells were added and the samples were incubated at room temperature for 30 minutes in dark. At the end of incubation period, CD8 antibody was added at 2 μL/1×10⁶ cells and the samples were incubated for another 30 minutes at room temperature in dark. At the end of incubation, FACS buffer was added to the samples and the samples were spun at 1300 rpm for 5 minutes. The pellet was resuspended in 5mL FACS buffer and the cells were spun at 1300 rpm for 5 minutes. The pellet was resuspended at a density of 3×10⁶/1 mL FACS buffer for the sorting.

Gating Strategy and Data Analysis

The cells were acquired on the BD FACS ARIA III flow cytometer and gated on FSC-A versus side scatter area (SSC-A). The cells high on FSC-A versus SSC-A were gated as lymphocytes. From the lymphocytes, single cells were gated on FSC-W versus FSC-H. Live cells were gated as brilliant violet 421 area (BV421-A) negative cells on the SSC-A versus BV421-A plot. The live cells were gated on SSC-A versus allophycocyanin-cyanine 7 area (APC-Cy7-A) and the positive cells on APC-Cy7 channel were marked as CD8⁺ cells. The dual mimic and mutant tetramer positive cells were gated by plotting PE-A versus APC-A within the CD8⁺ cells. The percentage of double positive cells for APC and PE is the frequency of mimic and mutant positive tetramer cells displayed in the gate. All gates were set using unstained samples and negative tetramers to eliminate non-specific binding. Mimic and mutant tetramers specific double positive were sorted into single cell/well in a 96 well plate containing cell lysis buffer for m-RNA preparation for NGS.

A summary of the CD8+ T cells (Table 7 and Table 8) positive for mimic and mutant tetramers for various donors is provided, below.

TABLE 7
Frequency (%) of double positive T cells
within CD8+ compartment of donors
		Mutant	Mutant	Mutant
		(SEQ ID	(SEQ ID	(SEQ ID
		NO: 9)	NO: 13)	NO: 18)
		Mimic	Mimic	Mimic
		(SEQ ID	(SEQ ID	(SEQ ID
	Donor ID	NO: 45)	NO: 59)	NO 68)
	17042765	0.14	0.13	0
	19054445	0.24	0.38	0.01
	19053796	0.1	0.06	0
	17042380	0.05	0.04	0.01
	19054456	0.01	0	0
	19054141	0.03	0.02	0
	19054183	0.01	0.01	0
	18047563	0.01	0.01	0

TABLE 8
Frequency (%) of double positive T cells
within CD8+ compartment of donors
	Mutant	Mutant	Mutant	Mutant	Mutant
	(SEQ ID	(SEQ ID	(SEQ ID	(SEQ ID	(SEQ ID
	NO: 2)	NO: 3)	NO: 16)	NO: 22)	NO: 23)
	Mimic	Mimic	Mimic	Mimic	Mimic
	(SEQ ID	(SEQ ID	(SEQ ID	(SEQ ID	(SEQ ID
Donor ID	NO: 29)	NO: 32)	NO: 64)	NO: 75)	NO: 78)
20061357	0.095	0.007	0.002	0.004	0.006
20001476	0.075	0.051	0.021	0.000	0.153
20062384	0.000	0.023	0.000	0.000	0.023
20062224	0.000	0.000	0.016	0.015	0.000
20061661	0.000	0.000	0.000	0.000	0.000
20001487	0.000	0.000	0.000	0.000	0.000
20061599	0.000	0.000	0.000	0.000	0.000

FIG. 1-FIG. 10 depict FACS plots used to determine the frequency of dual positive cells for mimic and mutant tetramers specific for mutant and mimic peptides represented by SEQ ID NO: 9 and SEQ ID NO: 45 (FIG. 1), mutant and mimic peptides represented by SEQ ID NO: 13 and SEQ ID NO: 59 (FIG. 2), mutant and mimic peptides represented by SEQ ID NO: 18 and SEQ ID NO: 68 (FIG. 3), negative APC tetramer (FIG. 4), negative PE tetramer (FIG. 5), mutant and mimic peptides represented by SEQ ID NO: 2 and SEQ ID NO: 29 (FIG. 6), mutant and mimic peptides represented by SEQ ID NO: 3 and SEQ ID NO: 32 (FIG. 7), mutant and mimic peptides represented by SEQ ID NO: 16 and SEQ ID NO: 64 (FIG. 8), mutant and mimic peptides represented by SEQ ID NO: 23 and SEQ ID NO: 78 (FIG. 9), and mutant and mimic peptides represented by SEQ ID NO: 22 and SEQ ID NO: 75 (FIG. 10).

Example 4

Identification of TCR Sequences that are Cross Reactive to Both Mutant and Mimic Peptides

The following donors were selected for individual single cell TCR sequencing based on a frequency of double positive T cells that was at least 2-fold higher than the background (negative tetramers): T-cells cells from donor 17042765 that were positive for mutant -mimic pairs a) SEQ ID NO: 9 and 45 and b) SEQ ID NO: 13 and 59; T-cells cells from donor 19054445 that were positive for mutant -mimic pairs a) SEQ ID NO: 9 and 45 and b) SEQ ID NO: 13 and 59. Also, T-cells from donor 19053796 were positive for following mutant-mimic pairs: a) SEQ ID NO: 9 and 45 b) SEQ ID NO: 13 and 59 and c) SEQ ID NO: 18 and 68.

Single cell TCR profiling was performed according to the methods described in the Takara Bio USA, SMARTer® Human scTCR a/b Profiling Kit User Manual, the entirety of which is incorporated herein by reference. In brief, single T cells were sorted into a 96-well plate. The cells were lysed, and first-strand synthesis was performed. cDNA was amplified by polymerase chain reaction (PCR) (16 cycles). The resultant cDNA was pooled and purified using Agencourt® AMPure® XP beads. Semi-nested PCR was used for TCR a/b amplification and sequencing library generation. The first TCR-specific PCR reaction was performed using 16 cycles and the second TCR-specific PCR reaction was performed using 14 cycles. The resultant cDNA was pooled and purified using Agencourt® AMPure® XP beads.

Library quality control was performed using Qubit quantification and TapeStation quality control. Libraries were pooled and sequencing was performed on 2x300 cycles V3 chemistry flow-cell on Illumina MiSeq.

Data analysis was performed by de-multiplexing using MiSeq Reported and checked for quality. TCR analysis was performed using Lymanalyzer and the results from individual single cell data were summarized by taking the best hit for TCR a/b for each cell data file.

Table 9 provides the donor number, sample name and the V(J) or V(D)J genes for the alpha and beta chains for TCRs positive for mutant-mimic pair SEQ ID NO: 9 and 45 tetramers. Each row represents an individual well in a 96 well plate.

TABLE 9
			TCR1	TCRB
	Row	Sample	V_Gene	J_Gene	V_Gene	D_Gene	J_Gene
Donor 19054445	1	S17	TRAV26-2*01	TRAJ37*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	2	S19	TRAV26-2*01	TRAJ37*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	3	S20	TRAV26-2*01	TRAJ37*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	4	S21	TRAV5*01	TRAJ41*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	5	S22	TRAV26-2*01	TRAJ37*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	6	S24	TRAV9-2*01	TRAJ18*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	7	S24	TRAV26-2*01	TRAJ37*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	8	S25	TRAV26-2*01	TRAJ37*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
Donor 19053796	9	S39	TRAV12-2*01	TRAJ15*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	10	S41	TRAV16*01	TRAJ41*01	TRBV20-1*05	TRBD2*02	TRBJ2-7*01
	11	S41	TRAV16*01	TRAJ41*01	TRBV20-1*05	TRBD2*02	TRBJ2-7*01
	12	S41	TRAV16*01	TRAJ41*01	TRBV20-1*05	TRBD2*02	TRBJ2-7*01
	13	S41	TRAV16*01	TRAJ41*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	14	S42	TRAV5*01	TRAJ41*01	TRBV11-1*01	TRBD2*02	TRBJ2-7*01
	15	S42	TRAV22*01	TRAJ37*01	TRBV15*02	TRBD2*02	TRBJ1-2*01
	16	S44	TRAV5*01	TRAJ41*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	17	S44	TRAV16*01	TRAJ41*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	18	S45	TRAV5*01	TRAJ41*01	TRBV11-1*01	TRBD2*02	TRBJ2-7*01
	19	S45	TRAV5*01	TRAJ41*01	TRBV11-1*01	TRBD2*02	TRBJ2-7*01
	20	S45	TRAV5*01	TRAJ41*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	21	S45	TRAV5*01	TRAJ41*01	TRBV11-1*01	TRBD2*02	TRBJ2-7*01
	22	S46	TRAV22*01	TRAJ37*01	TRBV5-6*01	TRBD2*02	TRBJ2-7*01
	23	S48	TRAV16*01	TRAJ41*01	TRBV15*02	TRBD2*02	TRBJ1-2*01
	24	S50	TRAV22*01	TRAJ37*01	TRBV12-4*01	TRBD2*02	TRBJ1-2*01

Table 10 provides the donor number, sample name and the V(J) or V(D)J genes for the alpha and beta chains for TCRs positive for mutant-mimic pair SEQ ID NO: 13 and 59 tetramers. Each row represents an individual well in a 96 well plate.

TABLE 10
			TCRA	TCRB
	Row	Sample	V_Gene	J_Gene	V_Gene	D_Gene	J_Gene
Donor 19054445	1	S27	TRAV24*01	TRAJ49*01	TRBV2*01	TRBD1*01	TRBJ2-4*01
	2	S28	TRAV4*01	TRAJ4*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	3	S29	TRAV9-2*01	TRAJ18*01	TRBV7-8*01	TRBD2*02	TRBJ1-5*01
	4	S29	TRAV22*01	TRAJ37*01	TRBV7-8*01	TRBD2*02	TRBJ1-5*01
	5	S29	TRAV24*01	TRAJ49*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	6	S29	TRAV24*01	TRAJ49*01	TRBV7-8*01	TRBD2*02	TRBJ1-5*01
	7	S30	TRAV13-1*01	TRAJ45*01	TRBV6-6*01	TRBD2*02	TRBJ1-4*01
	8	S30	TRAV22*01	TRAJ37*01	TRBV12-4*01	TRBD2*02	TRBJ2-7*01
	9	S30	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	10	S31	TRAV9-2*01	TRAJ18*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	11	S31	TRAV22*01	TRAJ37*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	12	S32	TRAV9-2*01	TRAJ18*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	13	S32	TRAV9-2*01	TRAJ18*01	TRBV7-8*01	TRBD2*02	TRBJ1-5*01
	14	S32	TRAV9-2*01	TRAJ18*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	15	S32	TRAV4*01	TRAJ4*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	16	S32	TRAV9-2*01	TRAJ18*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	17	S32	TRAV9-2*01	TRAJ18*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	18	S33	TRAV13-1*01	TRAJ45*01	TRBV6-6*01	TRBD2*02	TRBJ1-4*01
	19	S33	TRAV13-1*01	TRAJ45*01	TRBV6-6*01	TRBD2*02	TRBJ1-4*01
	20	S33	TRAV13-1*01	TRAJ45*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	21	S33	TRAV9-2*01	TRAJ18*01	TRBV7-8*01	TRBD2*02	TRBJ1-5*01
	22	S33	TRAV22*01	TRAJ37*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	23	S33	TRAV13-1*01	TRAJ45*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	24	S33	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	25	S34	TRAV4*01	TRAJ4*01	TRBV6-2*01, TRBV6-3*01	TRBD2*02	TRBJ2-2*01
	26	S34	TRAV4*01	TRAJ4*01	TRBV6-2*01, TRBV6-3*01	TRBD2*02	TRBJ2-2*01
	27	S34	TRAV9-2*01	TRAJ18*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	28	S34	TRAV4*01	TRAJ4*01	TRBV6-2*01, TRBV6-3*01	TRBD2*02	TRBJ2-2*01
	29	S34	TRAV4*01	TRAJ4*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	30	S36	TRAV22*01	TRAJ37*01	TRBV27*01	TRBD1*01	TRBJ2-1*01
	31	S38	TRAV22*01	TRAJ37*01	TRBV12-4*01	TRBD2*02	TRBJ2-7*01
	32	S38	TRAV22*01	TRAJ37*01	TRBV12-4*01	TRBD2*02	TRBJ2-7*01
	33	S38	TRAV22*01	TRAJ37*01	TRBV12-4*01	TRBD2*02	TRBJ2-7*01
	34	S38	TRAV22*01	TRAJ37*01	TRBV12-4*01	TRBD2*02	TRBJ2-7*01
Donor 19053796	35	S14	TRAV10*01	TRAJ40*01	TRBV7-9*03	TRBD2*01	TRBJ2-2*01
	36	S14	TRAV12-2*01	TRAJ43*01	TRBV18*01	TRBD1*01	TRBJ1-1*01
	37	S14	TRAV13-2*01	TRAJ9*01	TRBV7-9*03	TRBD2*01	TRBJ2-2*01
	38	S15	TRAV13-2*01	TRAJ15*01	TRBV11-2*01	TRBD1*01	TRBJ2-1*01
	39	S15	TRAV13-2*01	TRAJ15*01	TRBV5-1*01	TRBD2*02	TRBJ2-7*01
	40	S15	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	41	S15	TRAV13-2*01	TRAJ15*01	TRBV5-1*01	TRBD2*02	TRBJ2-7*01
	42	S16	TRAV10*01	TRAJ40*01	TRBV12-3*01	TRBD1*01	TRBJ2-1*01
	43	S16	TRAV10*01	TRAJ40*01	TRBV12-3*01	TRBD1*01	TRBJ2-1*01
	44	S16	TRAV10*01	TRAJ40*01	TRBV12-3*01	TRBD1*01	TRBJ2-1*01
	45	S16	TRAV10*01	TRAJ40*01	TRBV29-1*01	TRBD2*02	TRBJ2-7*01
	46	S16	TRAV10*01	TRAJ40*01	TRBV29-1*01	TRBD2*02	TRBJ2-7*01
	47	S17	TRAV10*01	TRAJ11*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	48	S17	TRAV10*01	TRAJ40*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	49	S17	TRAV12-2*01	TRAJ9*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	50	S18	TRAV10*01	TRAJ11*01	TRBV14*01	TRBD2*02	TRBJ2-5*01
	51	S18	TRAV10*01	TRAJ11*01	TRBV14*01	TRBD2*02	TRBJ2-5*01
	52	S18	TRAV10*01	TRAJ11*01	TRBV14*01	TRBD2*02	TRBJ2-5*01
	53	S18	TRAV25*01	TRAJ12*01	TRBV6-5*01	TRBD2*02	TRBJ2-7*01
	54	S18	TRAV10*01	TRAJ11*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	55	S19	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	56	S19	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	57	S20	TRAV10*01	TRAJ11*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	58	S20	TRAV10*01	TRAJ40*01	TRBV12-3*01	TRBD1*01	TRBJ2-1*01
	59	S22	TRAV10*01	TRAJ40*01	TRBV7-9*03	TRBD2*01	TRBJ2-2*01
	60	S23	TRAV10*01	TRAJ40*01	TRBV6-5*01	TRBD1*01	TRBJ2-3*01
	61	S38	TRAV12-2*01	TRAJ43*01	TRBV13*01	TRBD1*01	TRBJ2-2*01
	62	S38	TRAV13-2*01	TRAJ9*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	63	S38	TRAV12-2*01	TRAJ43*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	64	S38	TRAV12-2*01	TRAJ43*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	65	S38	TRAV12-2*01	TRAJ43*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	66	S39	TRAV26-1*01	TRAJ48*01	TRBV11-2*01	TRBD1*01	TRBJ2-7*01
	67	S40	TRAV13-1*01	TRAJ37*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	68	S42	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	69	S42	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	70	S43	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	71	S44	TRAV12-2*01	TRAJ43*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	72	S45	TRAV12-2*01	TRAJ9*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	73	S46	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	74	S46	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	75	S46	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	76	S46	TRAV12-2*01	TRAJ43*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	77	S46	TRAV9-2*01	TRAJ18*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	78	S46	TRAV9-2*01	TRAJ56*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	79	S46	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	80	S46	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	81	S47	TRAV9-2*01	TRAJ18*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	82	S47	TRAV26-2*01	TRAJ37*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	83	S48	TRAV8-4*01	TRAJ8*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	84	S48	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD1*01	TRBJ1-2*01
	85	S48	TRAV9-2*01	TRAJ18*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	86	S48	TRAV9-2*01	TRAJ56*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	87	S48	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01

Table 11 provides the donor number, sample name and the V(J) or V(D)J genes for the alpha and beta chains for TCRs positive for mutant-mimic pair SEQ ID NO: 18 and 68 tetramers. Each row represents an individual well in a 96 well plate.

TABLE 11
			TCRA	TCRB
	Row	Sample	V_Gene	J_Gene	V_Gene	D_Gene	J_Gene
Donor 19053796	1	S26	TRAV13-2*01	TRAJ9*01	TRBV3-1*01	TRBD1*01	TRBJ1-1*01
	2	S26	TRAV12-2*01	TRAJ43*01	TRBV18*01	TRBD1*01	TRBJ1-1*01
	3	S26	TRAV13-2*01	TRAJ9*01	TRBV3-1*01	TRBD1*01	TRBJ1-1*01
	4	S26	TRAV13-2*01	TRAJ9*01	TRBV18*01	TRBD1*01	TRBJ1-1*01
	5	S27	TRAV21*01	TRAJ26*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	6	S29	TRAV13-2*01	TRAJ9*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	7	S30	TRAV29/DV5*01	TRAJ26*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	8	S30	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	9	S32	TRAV13-2*01	TRAJ9*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	10	S32	TRAV12-3*01	TRAJ37*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	11	S34	TRAV13-2*01	TRAJ9*01	TRBV3-1*01	TRBD1*01	TRBJ1-1*01
	12	S34	TRAV12-3*01	TRAJ37*01	TRBV14*01	TRBD2*02	TRBJ2-7*01
	13	S35	TRAV12-3*01	TRAJ37*01	TRBV14*01	TRBD2*02	TRBJ2-7*01
	14	S35	TRAV6*01	TRAJ32*01	TRBV14*01	TRBD2*02	TRBJ2-7*01
	15	S35	TRAV13-2*01	TRAJ9*01	TRBV3-1*01	TRBD1*01	TRBJ1-1*01
	16	S35	TRAV12-3*01	TRAJ37*01	TRBV14*01	TRBD2*02	TRBJ2-7*01
	17	S35	TRAV12-3*01	TRAJ37*01	TRBV14*01	TRBD2*02	TRBJ2-7*01

Table 12 provides the donor number, sample name and the V(J) or V(D)J genes for the alpha and beta chains for TCRs positive for mutant-mimic pair SEQ ID NO: 3 and 32 tetramers. Each row represents an individual well in a 96 well plate.

TABLE 12
		TCRA	TCRB
	Row	V_Gene	J_Gene	V_Gene	D_Gene	J_Gene
Donor	1	TRAV26-2*01	TRAJ43*01	TRBV7-6*01	TRBD2*02	TRBJ1-4*01
	2	TRAV26-2*01	TRAJ43*01	TRBV7-6*01	TRBD2*02	TRBJ1-4*01
	3	TRAV8-4*01	TRAJ3*01	TRBV15*01	TRBD2*01	TRBJ2-1*01
	4	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	5	TRAV4*01	TRAJ20*01	TRBV4-1*01	TRBD1*01	TRBJ1-1*01
	6	TRAV4*01	TRAJ20*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	7	TRAV26-2*01	TRAJ43*01	TRBV5-1*01	TRBD1*01	TRBJ1-6*02
	8	TRAV12-3*01	TRAJ54*01	TRBV4-1*01	TRBD1*01	TRBJ2-1*01
	9	TRAV29/DV5*03	TRAJ41*01	TRBV5-4*04	TRBD1*01	TRBJ1-1*01
	10	TRAV26-2*01	TRAJ43*01	TRBV7-6*01	TRBD2*02	TRBJ1-4*01
	11	TRAV22*01	TRAJ32*01	TRBV2*01	TRBD1*01	TRBJ1-1*01
	12	TRAV22*01	TRAJ32*01	TRBV5-6*01	TRBD2*02	TRBJ2-7*01
	13	TRAV8-4*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	14	TRAV1-2*01	TRAJ28*01	TRBV6-2*01, TRBV6-3*01	TRBD1*01	TRBJ1-6*02
	15	TRAV12-2*01	TRAJ10*01	TRBV6-2*01, TRBV6-3*01	TRBD2*02	TRBJ2-7*01
	16	TRAV8-4*01, TRAV8-2*01	TRAJ3*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	17	TRAV8-4*01, TRAV8-2*01	TRAJ3*01	TRBV12-4*01, TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	18	TRAV23/DV6*01	TRAJ31*01	TRBV7-2*01	TRBD1*01	TRBJ2-1*01
	19	TRAV12-3*01	TRAJ24*01	TRBV12-4*01	TRBD2*02	TRBJ2-5*01
	20	TRAV26-2*01	TRAJ43*01	TRBV7-6*01	TRBD2*02	TRBJ1-4*01
	21	TRAV26-2*01	TRAJ43*01	TRBV7-6*01	TRBD2*02	TRBJ1-4*01
indicates data missing or illegible when filed

Table 13 provides the donor number, sample name and the V(J) or V(D)J genes for the alpha and beta chains for TCRs positive for mutant-mimic pair SEQ ID NO: 23 and 78 tetramers. Each row represents an individual well in a 96 well plate.

TABLE 13
		TCRA	TCRB
	Row	V_Gene	J_Gene	V_Gene	D_Gene	J_Gene
Donor 20001476	1	TRAV26-2*01	TRAJ43*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01
	2	TRAV29/DV5*01	TRAJ8*01	TRBV14*01	TRBD1*01	TRBJ2-1*01
	3	TRAV29/DV5*01	TRAJ8*01	TRBV14*01	TRBD1*01	TRBJ2-1*01
	4	TRAV29/DV5*01	TRAJ8*01	TRBV12-4*01,	TRBD2*02	TRBJ1-2*01
				TRBV12-3*01
	5	TRAV29/DV5*01	TRAJ8*01	TRBV12-3*01	TRBD2*02	TRBJ1-2*01

Table 14, Table 15,Table 16, Table 17 and Table 18 provide the CDR3 amino acid (aa) sequence, CDR3 nucleotide (nt) sequence and SEQ ID NOs for the mutant-mimic pair SEQ ID NO: 9 and SEQ ID NO: 45 tetramer positive TCRs identified in Table 9, the mutant-mimic pair SEQ ID NO: 13 and SEQ ID NO: 59 tetramer positive TCRs identified in Table 10, the mutant-mimic pair SEQ ID NO: 18 and SEQ ID NO: 68 tetramer positive TCRs identified in Table 11, the mutant-mimic pair SEQ ID NO: 3 and SEQ ID NO: 32 tetramer positive TCRs identified in Table 12, and the mutant-mimic pair SEQ ID NO: 23 and SEQ ID NO: 78 tetramer positive TCRs identified in Table 13, respectively. Each row in Table 14, Table 15, Table 16, Table 17 and Table 18 corresponds to the matching row in Table 9, Table 10, Table 11, Table 12 and Table 13, respectively, e.g. the V(J) or V(D)J genes in row 1 of Table 9 correspond to the CDR3 sequences in row 1 of Table 14, and so on.

TABLE 14
			TCRA	TCRB
				SEQ		SEQ		SEQ		SEQ
				ID		ID		ID		ID
	Row	Sample	CDR3 (aa)	NO	CDR3 (nt)	NO	CDR3 (aa)	NO	CDR3 (nt)	NO
Donor	1	S17	CVLLHKKTT	112	TGTGTACTACTGCAT	113	CASSFSTCS	114	TGTGCCAGCAGTTT	115
19054445			GKLIF		AAAAAAACAACAGGC		ANYGYTF		CTCGACCTGTTCGG
					AAACTAATCTTT				CTAACTATGGCTAC
									ACCTTC
	2	S19	CVLLHKKTT	112	TGTGTACTACTGCAT	113	CASSFSTCS	114	TGTGCCAGCAGTTT	115
			GKLIF		AAAAAAACAACAGGC		ANYGYTF		CTCGACCTGTTCGG
					AAACTAATCTTT				CTAACTATGGCTAC
									ACCTTC
	3	S70	CVLLHKKTT	117	TGTGTACTACTGCAT	113	CASSFSTCS	114	TGTGCCAGCAGTTT	115
			GKLIF		AAAAAAACAACAGGC		ANYGYTF		CTCGACCTGTTCGG
					AAACTAATCTTT				CTAACTATGGCTAC
									ACCTTC
	4	S2I	CAESPSGYA	116	TGTGCAGAGAGTCCT	117	CASSFSTCS	114	TGTGCCAGCAGTTT	115
			LNF		TCCGGGTATGCACTC		ANYGYTF		CTCGACCTGTTCGG
					AACTTC				CTAACTATGGCTAC
									ACCTTC
	5	S22	CVLLHKKTT	112	TGTGTACTACTGCAT	113	CASSFSTCS	114	TGTGCCAGCAGTTT	115
			GKLIF		AAAAAAACAACAGGC		ANYGYTF		CTCGACCTGTTCGG
					AAACTAATCTTT				CTAACTATGGCTAC
									ACCTTC
	6	S24	CALSEDRGS	118	TGTGCTCTGAGTGAA	119	CASSFSTCS	114	TGTGCCAGCAGTTT	115
			TLGRLYF		GACAGAGGCTCAACC		ANYGYTF		CTCGACCTGTTCGG
					CTGGGGAGGCTATAC				CTAACTATGGCTAC
					TTT				ACCTTC
	7	S24	CVLLHKKTT	112	TGTGTACTACTGCAT	113	CASSFSTCS	114	TGTGCCAGCAGTTT	115
			GKLIF		AAAAAAACAACAGGC		ANYGYTF		CTCGACCTGTTCGG
					AAACTAATCTTT				CTAACTATGGCTAC
									ACCTTC
	8	S25	CVLLHKKTT	112	TGTGTACTACTGCAT	113	CASSFSTCS	114	TGTGCCAGCAGTTT	115
			GKLIF		AAAAAAACAACAGGC		ANYGYTF		CTCGACCTGTTCGG
					AAACTAATCTTT				CTAACTATGGCTAC
									ACCTTC
Donor	9	S39	CAGLIGTAL	120	TGTGCCGGGTTAATA	121	CASSFSTCS	114	TGTGCCAGCAGTTT	115
19053796			IF		GGAACTGCTCTGATC		ANYGYTF		CTCGACCTGTTCGG
					TTT				CTAACTATGGCTAC
									ACCTTC
	10	S41	CALSRDSGY	122	TGTGCTCTAAGTAGG	123	CSAQGLAGE	124	TGCAGTGCCCAGGG	125
			ALNF		GATTCCGGGTATGCA		PIYEQYF		ACTAGCGGGTGAAC
					CTCAACTTC				CAATCTACGAGCAG
									TACTTC
	11	S41	CALSRDSGY	122	TGTGCTCTAAGTAGG	123	CSAQGLAGE	124	TGCAGTGCCCAGGG	125
			ALNF		GATTCCGGGTATGCA		PIYEQYF		ACTAGCGGGTGAAC
					CTCAACTTC				CAATCTACGAGCAG
									TACTTC
	12	S41	CALSRDSGY	122	TGTGCTCTAAGTAGG	123	CSAQGLAGE	124	TGCAGTGCCCAGGG	125
			ALNF		GATTCCGGGTATGCA		PIYEQYF		ACTAGCGGGTGAAC
					CTCAACTTC				CAATCTACGAGCAG
									TACTTC
	13	S41	CAQSRDSGY	126	TGTGCTCAAAGTAGG	127	CASSFSTCS	114	TGTGCCAGCAGTTT	115
			ALNF		GATTCCGGGTATGCA		ANYGYTF		CTCGACCTGTTCGG
					CTCAACTTC				CTAACTATGGCTAC
									ACCTTC
	14	S42	CAESPSGYA	116	TGTGCAGAGAGTCCT	117	CASSLKLAP	128	TGTGCCAGCAGCTT	129
			LNF		TCCGGGTATGCACTC		YEQYF		GAAACTAGCCCCCT
					AACTTC				ACGAGCAGTACTTC
	15	S42	CTFPLPRPQ	130	TGTACATTTCCTCTT	131	CATSFPDLY	134	TGTGCCACCAGCTT	135
			TQAFISVLS		CCCAGACCACAGACT		GYTF		CCCGGACCTCTATG
			RTSASNTGK		CAGGCGTTTATTTCT				GCTACACCTTC
			LIF		GTGCTGTCCCGAACT
					TCAGCTAGCAACACA
					GGCAAACTAATCTTT
	16	S44	CAESPSGYA	116	TGTGCAGAGAGTCCT	117	CASSFSTCS	114	TGTGCCAGCAGTTT	115
			LNF		TCCGGGTATGCACTC		ANYGYTF		CTCGACCTGTTCGG
					AACTTC				CT17AACTATGGCT
									ACACCTTC
	17	S44	CALSRDSGY	122	TGTGCTCTAAGTAGG	123	CASSFSTCS	114	TGTGCCAGCAGTTT	115
			ALNF		GATTCCGGGTATGCA		ANYGYTF		CTCGACCTGTTCGG
					CTCAACTTC				CTAACTATGGCTAC
									ACCTTC
	18	S45	CAESPSGYA	116	TGTGCAGAGAGTCCT	117	CASSLKLAP	128	TGTGCCAGCAGCTT	129
			LNF		TCCGGGTATGCACTC		YEQYF		GAAACTAGCCCCCT
					AACTTC				ACGAGCAGTACTTC
	19	S45	CAESPSGYA	116	TGTGCAGAGAGTCCT	117	CASSLKLAP	128	TGTGCCAGCAGCTT	129
			LNF		TCCGGGTATGCACTC		YEQYF		GAAACTAGCCCCCT
					AACTTC				ACGAGCAGTACTTC
	20	S45	CAESPSGYA	116	TGTGCAGAGAGTCCT	117	CASSFSTCS	114	TGTGCCAGCAGTTT	115
			LNF		TCCGGGTATGCACTC		ANYGYTF		CTCGACCTGTTCGG
					AACTTC				CTAACTATGGCTAC
									ACCTTC
	21	S45	CAESPSGYA	116	TGTGCAGAGAGTCCT	117	CASSLKLAP	128	TGTGCCAGCAGCTT	129
			LNF		TCCGGGTATGCACTC		YEQYF		GAAACTAGCCCCCT
					AACTTC				ACGAGCAGTACTTC
	22	S46	CTFPLPRPQ	130	TGTACATTTCCTCTT	131	CASSESTYE	132	TGTGCCAGCAGCGA	133
			TQAFISVLS		CCCAGACCACAGACT		QYF		GAGTACCTACGAGC
			RTSASNTGK		CAGGCGTTTATTTCT				AGTACTTC
			LIF		GTGCTGTCCCGAACT
					TCAGCTAGCAACACA
					GGCAAACTAATCTTT
	23	S48	CALSRDSGY	122	TGTGCTCTAAGTAGG	123	CATSFPDLY	134	TGTGCCACCAGCTT	135
			ALNF		GATTCCGGGTATGCA		GYTF		CCCGGACCTCTATG
					CTCAACTTC				GCTACACCTTC
	24	S50	CTFPLPRPQ	130	TGTACATTTCCTCTT	131	CASSFSTCS	114	TGTGCCAGCAGTTT	115
			TQAFISVLS		CCCAGACCACAGACT		ANYGYTF		CTCGACCTGTTCGG
			RTSASNTGK		CAGGCGTTTATTTCT				CTAACTATGGCTAC
			LIF		GTGCTGTCCCGAACT				ACCTTC
					TCAGCTAGCAACACA
					GGCAAACTAATCTTT

TABLE 15
			TCRA	TCRB
				SEQ		SEQ		SEQ		SEQ
		Sam-		ID		ID		ID		ID
	Row	ple	CDR3 (aa)	NO	CDR3 (nt)	NO	CDR3 (aa)	NO	CDR3 (nt)	NO
Donor	1	S27	CARNTGNQFY	136	TGTGCCCGGA	137	CASRSGVLLA	138	TGTGCCAGCA	139
19054445			F		ACACCGGTAA		KNIQYF		GATCGGGTGT
					CCAGTTCTAT				ACTACTAGCC
					TTT				AAAAACATTC
									AGTACTTC
	2	S28	CLVGDRGLMF	140	TGCCTCGTGG	141	CASSFSTCSA	114	TGTGCCAGCA	115
			SGGYNKLIF		GTGACAGGGG		NYGYTF		GTTTCTCGAC
					ACTCATGTTT				CTGTTCGGCT
					TCTGGTGGCT				AACTATGGCT
					ACAATAAGCT				ACACCTTC
					GATTTTT
	3	S29	CALSEDRGST	118	TGTGCTCTGA	119	CASSSLSNQP	142	TGTGCCAGCA	143
			LGRLYF		GTGAAGACAG		QHF		GCTCGTTGAG
					AGGCTCAACC				CAATCAGCCC
					CTGGGGAGGC				CAGCATTTT
					TATACTTT
	4	S29	CTFPLPRPQT	130	TGTACATTTC	131	CASSSLSNQP	142	TGTGCCAGCA	143
			QAFISVLSRT		CTCTTCCCAG		QHF		GCTCGTTGAG
			SASNTGKLIF		ACCACAGACT				CAATCAGCCC
					CAGGCGTTTA				CAGCATTTT
					TTTCTGTGCT
					GTCCCGAACT
					TCAGCTAGCA
					ACACAGGCAA
					ACTAATCTTT
	5	S29	CARNTGNQFY	136	TGTGCCCGGA	137	CASSFSTCSA	114	TGTGCCAGCA	115
			F		ACACCGGTAA		NYGYTF		GTTTCTCGAC
					CCAGTTCTAT				CTGTTCGGCT
					TTT				AACTATGGCT
									ACACCTTC
	6	S29	CARNTGNQFY	136	TGTGCCCGGA	137	CASSSLSNQP	142	TGTGCCAGCA	143
			F		ACACCGGTAA		QHF		GCTCGTTGAG
					CCAGTTCTAT				CAATCAGCCC
					TTT				CAGCATTTT
	7	S30	CAARGGADGL	144	TGTGCAGCAC	145	CASSYYGQGG	146	TGTGCCAGCA	147
			TF		GAGGAGGTGC		EKLFF		GTTACTATGG
					TGACGGACTC				ACAGGGGGGA
					ACCTTT				GAAAAACTGT
									TTTTT
	8	S30	CTFPLPRPQT	148	TGTACATTTC	149	CASSSDRVYE	150	TGTGCCAGCA	151
			QAFISVLSRT		CTCTTCCCAG		QYF		GTTCCGACCG
			AASNTGKLIF		ACCACAGACT				AGTTTACGAG
					CAGGCGTTTA				CAGTACTTC
					TTTCTGTGCT
					GTCCCGAACT
					GCAGCTAGCA
					ACACAGGCAA
					ACTAATCTTT
	9	S30	CAVSDLEPNS	152	TGTGCTGTGA	153	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTCGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T
	10	S31	CALSEDRGST	118	TGTGCTCTGA	119	CASSFSTCSA	114	TGTGCCAGCA	115
			LGRLYF		GTGAAGACAG		NYGYTF		GTTTCTCGAC
					AGGCTCAACC				CTGTTCGGCT
					CTGGGGAGGC				AACTATGGCT
					TATACTTT				ACACCTTC
	11	S31	CTFPLPRPQT	130	TGTACATTTC	131	CASSFSTCSA	114	TGTGCCAGCA	115
			QAFISVLSRT		CTCTTCCCAG		NYGYTF		GTTTCTCGAC
			SASNTGKLIF		ACCACAGACT				CTGTTCGGCT
					CAGGCGTTTA				AACTATGGCT
					TTTCTGTGCT				ACACCTTC
					GTCCCGAACT
					TCAGCTAGCA
					ACACAGGCAA
					ACTAATCTTT
	12	S32	CALSEDRGST	118	TGTGCTCTGA	119	CASSFSTCSA	114	TGTGCCAGCA	115
			LGRLYF		GTGAAGACAG		NYGYTF		GTTTCTCGAC
					AGGCTCAACC				CTGTTCGGCT
					CTGGGGAGGC				AACTATGGCT
					TATACTTT				ACACCTTC
	13	S32	CALSEDRGST	118	TGTGCTCTGA	119	CASSSLSNQP	142	TGTGCCAGCA	143
			LGRLYF		GTGAAGACAG		QHF		GCTCGTTGAG
					AGGCTCAACC				CAATCAGCCC
					CTGGGGAGGC				CAGCATTTT
					TATACTTT
	14	S32	CALSEDRGST	118	TGTGCTCTGA	119	CASSFSTCSA	114	TGTGCCAGCA	115
			LGRLYF		GTGAAGACAG		NYGYTF		GTTTCTCGAC
					AGGCTCAAC				CTGTTCGGCT
					CCTGGGGAGG				AACTATGGCT
					CTATACTTT				ACACCTTC
	15	S32	CLVGDRGLMF	140	TGCCTCGTGG	141	CASSFSTCSA	114	TGTGCCAGCA	115
			SGGYNKLIF		GTGACAGGGG		NYGYTF		GTTTCTCGAC
					ACTCATGTTT				CTGTTCGGCT
					TCTGGTGGCT				AACTATGGCT
					ACAATAAGCT				ACACCTTC
					GATTTTT
	16	S32	CALREDRGST	154	TGTGCTCTGC	155	CKPISGHNSL	156	TGTAAACCAA	157
			LGRLYF		GTGAAGACAG		FWYRQTMMRG		TTTCAGGCCA
					AGGCTCAACC		LELLIYFNNN		CAACTCCCTT
					CTGGGGAGGC		VPIDDSGMPE		TTCTGGTACA
					TATACTTT		DRFSAKMPNA		GACAGACCAT
							SFSTLKIQPS		GATGCGGGGA
							EPRDSAVYFY		CTGGAGTTGC
							ASSFSTCSAN		TCATTTACTT
							YGYTF		TAACAACAAC
									GTTCCGATAG
									ATGATTCAGG
									GATGCCCGAG
									GATCGATTCT
									CAGCTAAGAT
									GCCTAATGCA
									TCATTCTCCA
									CTCTGAAGAT
									CCAGCCCTCA
									GAACCCAGGG
									ACTCAGCTGT
									GTACTTCTAT
									GCCAGCAGTT
									TCTCGACCTG
									TTCGGCTAAC
									TATGGCTACA
									CCTTC
	17	S32	CALSEDRGST	118	TGTGCTCTGA	119	CASSFSTCSA	114	TGTGCCAGCA	115
			LGRLYF		GTGAAGACAG		NYGYTF		GTTTCTCGAC
					AGGCTCAACC				CTGTTCGGCT
					CTGGGGAGGC				AACTATGGCT
					TATACTTT				ACACCTTC
	18	S33	CAARGGADGL	144	TGTGCAGCAC	145	CASSYYGQGG	146	TGTGCCAGCA	147
			TF		GAGGAGGTGC		EKLFF		GTTACTATGG
					TGACGGACTC				ACAGGGGGGA
					ACCTTT				GAAAAACTGT
									TTTTT
	19	S33	CAARGGADGL	144	TGTGCAGCAC	145	CASSYYGQGG	146	TGTGCCAGCA	147
			TF		GAGGAGGTGC		EKLFF		GTTACTATGG
					TGACGGACTC				ACAGGGGGGA
					ACCTTT				GAAAAACTGT
									TTTTT
	20	S33	CAARGGADGL	144	TGTGCAGCAC	145	CASSFSTCSA	114	TGTGCCAGCA	115
			TF		GAGGAGGTGC		NYGYTF		GTTTCTCGAC
					TGACGGACTC				CTGTTCGGCT
					ACCTTT				AACTATGGCT
									ACACCTTC
	21	S33	CALSEDRGST	118	TGTGCTCTGA	119	CASSSLSNQP	142	TGTGCCAGCA	143
			LGRLYF		GTGAAGACAG		QHF		GCTCGTTGAG
					AGGCTCAACC				CAATCAGCCC
					CTGGGGAGGC				CAGCATTTT
					TATACTTT
	22	S33	CTFPLPRPQT	148	TGTACATTTC	149	CASSFSTCSA	114	TGTGCCAGCA	115
			QAFISVLSRT		CTCTTCCCAG		NYGYTF		GTTTCTCGAC
			AASNTGKLIF		ACCACAGACT				CTGTTCGGCT
					CAGGCGTTTA				AACTATGGCT
					TTTCTGTGCT				ACACCTTC
					GTCCCGAACT
					GCAGCTAGCA
					ACACAGGCAA
					ACTAATCTTT
	23	S33	CAARGGADGL	144	TGTGCAGCAC	145	CKPISGHNSL	158	TGTAAACCAA	159
			TF		GAGGAGGTGC		FWYRQTMMRG		TTTCAGGCCA
					TGACGGACTC		LELLIYFNNN		CAACTCCCTT
					ACCTTT		VPIDDSGMPE		TTCTGGTACA
							DRFSAKMPNA		GACAGACCAT
							SFSTLKIQPS		GATGCGGGGA
							EPRDSAVYFG		CTGGAGTTGC
							ASSFSTCSAN		TCATTTACTT
							YGYTF		TAACAACAAC
									GTTCCGATAG
									ATGATTCAGG
									GATGCCCGAG
									GATCGATTCT
									CAGCTAAGAT
									GCCTAATGCA
									TCATTCTCCA
									CTCTGAAGAT
									CCAGCCCTCA
									GAACCCAGGG
									ACTCAGCTGT
									GTACTTCGGT
									GCCAGCAGTT
									TCTCGACCTG
									TTCGGCTA
									ACTATGGCTA
									CACCTTC
	24	S33	CAVSDLEPNS	152	TGTGCTGTGA	153	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTCGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T
	25	S34	CLVGDRGLMF	140	TGCCTCGTGG	141	CATLAGSTNT	160	TGTGCCACCC	161
			SGGYNKLIF		GTGACAGGGG		GELFF		TTGCCGGGTC
					ACTCATGTTT				TACGAACACC
					TCTGGTGGCT				GGGGAGCTGT
					ACAATAAGCT				TTTTT
					GATTTTT
	26	S34	CLVGDRGLMF	140	TGCCTCGTGG	141	CATLAGSTNT	160	TGTGCCACCC	161
			SGGYNKLIF		GTGACAGGGG		GELFF		TTGCCGGGTC
					ACTCATGTTT				TACGAACACC
					TCTGGTGGCT				GGGGAGCTGT
					ACAATAAGCT				TTTTT
					GATTTTr
	27	S34	CALSEDRGST	118	TGTGCTCTGA	119	CASSFSTCSA	114	TGTGCCAGCA	115
			LGRLYF		GTGAAGACAG		NYGYTF		GTTTCTCGAC
					AGGCTCAACC				CTGTTCGGCT
					CTGGGGAGGC				AACTATGGCT
					TATACTTT				ACACCTTC
	28	S34	CLVGDRGLMF	140	TGCCTCGTGG	141	CATLAGSTNT	160	TGTGCCACCC	161
			SGGYNKLIF		GTGACAGGGG		GELFF		TTGCCGGGTC
					ACTCATGTTT				TACGAACACC
					TCTGGTGGCT				GGGGAGCTGT
					ACAATAAGCT				TTTTT
					GATTTTT
	29	S34	CLVGDRGLMF	140	TGCCTCGTGG	141	CASSFSTCSA	114	TGTGCCAGCA	115
			SGGYNKLIF		GTGACAGGGG		NYGYTF		GTTTCTCGAC
					ACTCATGTTT				CTGTTCGGCT
					TCTGGTGGCT				AACTATGGCT
					ACAATAAGCT				ACACCTTC
					GATTTTT
	30	S36	CTFPLPRPQT	130	TGTACATTTC	131	CASSLSMNRV	162	TGTGCCAGCA	163
			QAFISVLSRT		CTCTTCCCAG		KNEQFF		GTTTATCCAT
			SASNTGKLIF		ACCACAGACT				GAACAGGGTT
					CAGGCGTTTA				AAGAATGAGC
					TTTCTGTGCT				AGTTCTTC
					GTCCCGAACT
					TCAGCTAGCA
					ACACAGGCAA
					ACTAATCTTT
	31	S38	CTFPLPRPQT	130	TGTACATTTC	131	CASSSDRVYE	150	TGTGCCAGCA	151
			QAFISVLSRT		CTCTTCCCAG		QYF		GTTCCGACCG
			SASNTGKLIF		ACCACAGACT				AGTTTACGAG
					CAGGCGTTTA				CAGTACTTC
					TTTCTGTGCT
					GTCCCGAACT
					TCAGCTAGCA
					ACACAGGCAA
					ACTAATCTTT
	32	S38	CTFPLPRPQT	130	TGTACATTTC	131	CASSSDRVYE	150	TGTGCCAGCA	151
			QAFISVLSRT		CTCTTCCCAG		QYF		GTTCCGACCG
			SASNTGKLIF		ACCACAGACT				AGTTTACGAG
					CAGGCGTTTA				CAGTACTTC
					TTTCTGTGCT
					GTCCCGAACT
					TCAGCTAGCA
					ACACAGGCAA
					ACTAATCTTT
	33	S38	CTFPLPRPQT	130	TGTACATTTC	131	CASSSDRVYE	150	TGTGCCAGCA	151
			QAFISVLSRT		CTCTTCCCAG		QYF		GTTCCGACCG
			SASNTGKLIF		ACCACAGACT				AGTTTACGAG
					CAGGCGTTTA				CAGTACTTC
					TTTCTGTGCT
					GTCCCGAACT
					TCAGCTAGCA
					ACACAGGCAA
					ACTAATCTTT
	34	S38	CTFPLPRPQT	130	TGTACATTTC	131	CASSSDRVYE	150	TGTGCCAGCA	151
			QAFISVLSRT		CTCTTCCCAG		QYF		GTTCCGACCG
			SASNTGKLIF		ACCACAGACT				AGTTTACGAG
					CAGGCGTTTA				CAGTACTTC
					TTTCTGTGCT
					GTCCCGAACT
					TCAGCTAGCA
					ACACAGGCAA
					ACTAATCTTT
Donor	35	S14	CVVSERTSGT	164	TGTGTGGTGA	165	CASSLGGPGE	166	TGTGCCAGCA	167
19053796			YKYIF		GCGAAAGGAC		LFF		GCCTAGGGGG
					CTCAGGAACC				ACCCGGGGAG
					TACAAATACA				CTGTnTTT
					TCTTT
	36	S14	CREHGDDMRF	168	TGCCGTGAAC	169	CASSPLRDNT	170	TGTGCCAGCT	171
					ATGGCGATGA		EAFF		CACCACTTCG
					CATGCGCTTT				GGACAACACC
									GAAGCTTTCT
									TT
	37	S14	CKLQLLNLET	172	TGCAAATTGC	173	CASSLGGPGE	166	TGTGCCAGCA	167
			QLSTFVPENT		AGCTACTCAA		LFF		GCCTAGGGGG
			GGFKTIF		CCTGGAGACT				ACCCGGGGAG
					CAGCTGTCTA				CTGTTTTTT
					CTTTT
					GTGCCTGAAA
					ATACTGGAGG
					CTTCAAAACT
					ATCTTT
	38	S15	CKLQLLNLET	174	TGCAAATTGC	175	CASHLGTGAY	176	TGTGCCAGCC	177
			QLSTFVQRQT		AGCTACTCAA		NEQFF		ATTTAGGGAC
			QNQAGTALIF		CCTGGAGACT				AGGGGCTTAC
					CAGCTGTCTA				AATGAGCAGT
					CTTTTGTGCA				TCTTC
					GAGACAAACG
					CAAAACCAGG
					CAGGAACTGC
					TCTGATCTTT
	39	S15	CKLQLLNLET	174	TGCAAATTGC	175	CASSLDPESW	178	TGCGCCAGCA	179
			QLSTFVQRQT		AGCTACTCAA		GPSYEQYF		GCTTGGATCC
			QNQAGTALIF		CCTGGAGACT				CGAGAGCTGG
					CAGCTGTCTA				GGACCCTCCT
					CTTTTGTGCA				ACGAGCAGTA
					GAGACAAACG				CTTC
					CAAAACCAGG
					CAGGAACTGC
					TCTGATCTTT
	40	S15	CAVSDLEPNS	152	TGTGCTGTGA	153	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTCGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T
	41	S15	CKLQLLNLET	174	TGCAAATTGC	175	CASSLDPESW	178	TGCGCCAGCA	179
			QLSTFVQRQT		AGCTACTCAA		GPSYEQYF		GCTTGGATCC
			QNQAGTALIF		CCTGGAGACT				CGAGAGCTGG
					CAGCTGTCTA				GGACCCTCCT
					CTTTTGTGCA				ACGAGCAGTA
					GAGACAAACG				CTTC
					CAAAACCAGG
					CAGGAACTGC
					TCTGATCTTT
	42	S16	CVVSERTSGT	164	TGTGTGGTGA	165	CASSFGSYHN	180	TGTGCCAGCA	181
			YKYIF		GCGAAAGGAC		EQFF		GTTTTGGCTC
					CTCAGGAACC				TTATCACAAT
					TACAAATACA				GAGCAGTTCT
					TCTTT				TC
	43	S16	CWSERTSGTY	164	TGTGTGGTGA	165	CASSFGSYHN	180	TGTGCCAGCA	181
			KYIF		GCGAAAGGAC		EQFF		GTTTTGGCTC
					CTCAGGAACC				TTATCACAAT
					TACAAATACA				GAGCAGTTCT
					TCTTT				TC
	44	S16	CVVSERTSGT	164	TGTGTGGTGA	165	CASSFGSYHN	180	TGTGCCAGCA	181
			YKYIF		GCGAAAGGAC		EQFF		GTTTTGGCTC
					CTCAGGAACC				TTATCACAAT
					TACAAATACA				GAGCAGTTCT
					TCTTT				TC
	45	S16	CWSERTSGTY	164	TGTGTGGTGA	165	CSVVGGVTYE	182	TGCAGCGTTG	183
			KYIF		GCGAAAGGAC		QYF		TAGGGGGCGT
					CTCAGGAACC				TACCTACGAG
					TACAAATACA				CAGTACTTC
					TCTTT
	46	S16	CVVSERTSGT	164	TGTGTGGTGA	165	CSVVGGVTYE	182	TGCAGCGTTG	183
			YKYIF		GCGAAAGGAC		QYF		TAGGGGGCGT
					CTCAGGAACC				TACCTACGAG
					TACAAATACA				CAGTACTTC
					TCTTT
	47	S17	CGERRNSGYS	184	TGTGGTGAGC	185	CASSFSTCSA	114	TGTGCCAGCA	115
			TLTF		GCAGGAATTC		NYGYTF		GTTTCTCGAC
					AGGATACAGC				CTGTTCGGCT
					ACCCTCACCT				AACTATGGCT
					TT				ACACCTTC
	48	S17	CVVSERTSGT	164	TGTGTGGTGA	165	CASSFSTCSA	114	TGTGCCAGCA	115
			YKYIF		GCGAAAGGAC		NYGYTF		GTTTCTCGAC
					CTCAGGAACC				CTGTTCGGCT
					TACAAATACA				AACTATGGCT
					TCTTT				ACACCTTC
	49	S17	CALGGFKTIF	186	TGTGCCTTGG	187	CASSFSTCSA	114	TGTGCCAGCA	115
					GAGGCTTCAA		NYGYTF		GTTTCTCGAC
					AACTATCTTT				CTGTTCGGCT
									AACTATGGCT
									ACACCTTC
	50	SI8	CGERRNSGYS	184	TGTGGTGAGC	185	CASSQDRETQ	188	TGTGCCAGCA	189
			TLTF		GCAGGAATTC		YF		GCCAAGATAG
					AGGATACAGC				GGAGACCCAG
					ACCCTCACCT				TACTTC
					TT
	51	S18	CGERRNSGYS	184	TGTGGTGAGC	185	CASSQDRETQ	188	TGTGCCAGCA	189
			TLTF		GCAGGAATTC		YF		GCCAAGATAG
					AGGATACAGC				GGAGACCCAG
					ACCCTCACCT				TACTTC
	52	S18	CGERRNSGYS	184	TGTGGTGAGC	185	CASSQDRETQ	188	TGTGCCAGCA	189
			TLTF		GCAGGAATTC		YF		GCCAAGATAG
					AGGATACAGC				GGAGACCCAG
					ACCCTCACCT				TACTTC
					TT
	53	S18	CAGHAITRPM	190	TGTGCAGGGC	191	CASSYGSPAQ	192	TGTGCCAGCA	193
			DSSYKLIF		ACGCGATAAC		DEQYF		GTTACGGGTC
					CCGACCGATG				CCCCGCTCAG
					GATAGCAGCT				GACGAGCAGT
					ATAAATTGAT				ACTTC
					CTTC
	54	S18	CGERRNSGYS	194	TGTGGTGAGC	195	CASSFSTCSA	114	TGTGCCAGCA	115
			NLTF		GCAGGAATTC		NYGYTF		GTTTCTCGAC
					AGGATACAGC				CTGTTCGGCT
					AACCTCACCT				AACTATGGCT
					TT				ACACCTTC
	55	S19	CAVSDLEPNS	152	TGTGCTGTGA	153	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTCGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T
	56	S19	CAVSDLEPNS	152	TGTGCTGTGA	153	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTCGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T
	57	S20	out_of_	N/A	TGTTTCCCTG	196	CASSFSTCSA	114	TGTGCCAGCA	115
			frame		ACAATCATGA		NYGYTF		GTTTCTCGAC
					CTTTCAGTGA				CTGTTCGGCT
					GAACACAAAG				AACTATGGCT
					TCGAACGGAA				ACACCTTC
					GAGATACAGC
					AACACTGGAG
					GAAGACACAA
					AGCAAAGATC
					AAGGCACAAC
					ACAGCCTCCC
					AGCTCAGCGA
					TAGAGCCTCC
					TACATCTGGG
					TGATGAGCGA
					AGGAATAGAG
					GGTACAGCAA
					CCTCATCTTT
	58	S20	CWSERTSGTY	164	TGTGTGGTGA	165	CASSFGSYHN	180	TGTGCCAGCA	181
			KYIF		GCGAAAGGAC		EQFF		GTTTTGGCTC
					CTCAGGAACC				TTATCACAAT
					TACAAATACA				GAGCAGTTCT
					TCTTT				TC
	59	S22	CVVSERTSGT	164	TGTGTGGTGA	165	CASSLGGPGE	166	TGTGCCAGCA	167
			YKYIF		GCGAAAGGAC		LFF		GCCTAGGGGG
					CTCAGGAACC				ACCCGGGGAG
					TACAAATACA				CTGTTTTTT
					TCTTT
	60	S23	CVVSERTSGT	164	TGTGTGGTGA	165	CASSYGQLAD	197	TGTGCCAGCA	198
			YKYIF		GCGAAAGGAC		TQYF		GTTACGGCCA
					CTCAGGAACC				GTTGGCCGAT
					TACAAATACA				ACGCAGTATT
					TCTTT				TT
Donor	61	S38	CREHGDDMRF	168	TGCCGTGAAC	169	CASSSTGTGN	199	TGTGCCAGCA	200
17042765					ATGGCGATGA		TGELFF		GTTCAACCGG
					CATGCGCTTT				GACAGGGAAC
									ACCGGGGAGC
									TGTTTTTT
	62	S38	CKLQLLNLET	172	TGCAAATTGC	173	CASSFSTCSA	114	TGTGCCAGCA	115
			QLSTFVPENT		AGCTACTCAA		NYGYTF		GTTTCTCGAC
			GGFKTIF		CCTGGAGACT				CTGTTCGGCT
					CAGCTGTCTA				AACTATGGCT
					CTTTTGTGCC				ACACCTTC
					TGAAAATACT
					GGAGGCTTCA
					AAACTATCTT
					T
	63	S38	CREHGDDMRF	168	TGCCGTGAAC	169	CASSFSTCSA	114	TGTGCCAGCA	115
					ATGGCGATGA		NYGYTF		GTTTCTCGAC
					CATGCGCTTT				CTGTTCGGCT
									AACTATGGCT
									ACACCTTC
	64	S38	CREHGDDMRF	168	TGCCGTGAAC	169	CASSFSTCSA	114	TGTGCCAGCA	115
					ATGGCGATGA		NYGYTF		GTTTCTCGAC
					CATGCGCTTT				CTGTTCGGCT
									AACTATGGCT
									ACACCTTC
	65	S38	CREHGDDMRF	168	TGCCGTGAAC	169	CASSFSTCSA	114	TGTGCCAGCA	115
					ATGGCGATGA		NYGYTF		GTTTCTCGAC
					CATGCGCTTT				CTGTTCGGCT
									AACTATGGCT
									ACACCTTC
	66	S39	CIVGRDFGNE	201	TGCATCGTGG	202	CASSLERAGA	203	TGTGCCAGCA	204
			KLTF		GCCGGGACTT		YEQYF		GCTTAGAGCG
					TGGAAATGAG				GGCAGGGGCC
					AAATTAACCT				TACGAGCAGT
					TT				ACTTC
	67	S40	CAASIPARSN	205	TGTGCAGCAA	206	CASSFSTCSA	114	TGTGCCAGCA	115
			TGKLIF		GTATACCCGC		NYGYTF		GTTTCTCGAC
					CAGGAGCAAC				CTGTTCGGCT
					ACAGGCAAAC				AACTATGGCT
					TAATCTTT				ACACCTTC
	68	S42	CAVSDLEPNS	152	TGTGCTGTGA	153	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTCGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T
	69	S42	CAVSDLEPNS	152	TGTGCTGTGA	153	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTCGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T
	70	S43	CAVSDLEPNS	152	TGTGCTGTGA	153	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTCGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T
	71	S44	CREHGDDMRF	168	TGCCGTGAAC	169	CASSFSTCSA	114	TGTGCCAGCA	115
					ATGGCGATGA		NYGYTF		GTTTCTCGAC
					CATGCGCTTT				CTGTTCGGCT
									AACTATGGCT
									ACACCTTC
	72	S45	CALGGFKTIF	186	TGTGCCTTGG	187	CASSFSTCSA	114	TGTGCCAGCA	115
					GAGGCTTCAA		NYGYTF		GTTTCTCGAC
					AACTATCTTT				CTGTTCGGCT
									AACTATGGCT
									ACACCTTC
	73	S46	CAVSDLEPNS	152	TGTGCTGTGA	153	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTCGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T
	74	S46	CAVSDLEPNS	152	TGTGCTGTGA	153	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTCGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T
	75	S46	CAVSDLEPNS	152	TGTGCTGTGA	153	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTCGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T
	76	S46	CREHGDDMRF	168	TGCCGTGAAC	169	CASSFSTCSA	114	TGTGCCAGCA	115
					ATGGCGATGA		NYGYTF		GTTTCTCGAC
					CATGCGCTTT				CTGTTCGGCT
									AACTATGGCT
									ACACCTTC
	77	S46	CALSEDRGST	118	TGTGCTCTGA	119	CASSFSTCSA	114	TGTGCCAGCA	115
			LGRLYF		GTGAAGACAG		NYGYTF		GTTTCTCGAC
					AGGCTCAACC				CTGTTCGGCT
					CTGGGGAGGC				AACTATGGCT
					TATACTTT				ACACCTTC
	78	S46	CALQAGGGAN	207	TGTGCTCTGC	208	CASSFSTCSA	114	TGTGCCAGCA	115
			SKLTF		AAGCGGGAGG		NYGYTF		GTTTCTCGAC
					TGGAGCCAAT				CTGTTCGGCT
					AGTAAGCTGA				AACTATGGCT
					CATTT				ACACCTTC
	79	S46	CAVSDLEPNS	152	TGTGCTGTGA	153	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTCGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T
	80	S46	CAVSDLEPNS	152	TGTGCTGTGA	209	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTGGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T
	81	S47	CALSEDRGST	118	TGTGCTCTGA	119	CASSFSTCSA	114	TGTGCCAGCA	115
			LGRLYF		GTGAAGACAG		NYGYTF		GTTTCTCGAC
					AGGCTCAACC				CTGTTCGGCT
					CTGGGGAGGC				AACTATGGCT
					TATACTTT				ACACCTTC
	82	S47	CVLLHKKTTG	112	TGTGTACTAC	113	CASSFSTCSA	114	TGTGCCAGCA	115
			KLIF		TGCATAAAAA		NYGYTF		GTTTCTCGAC
					AACAACAGGC				CTGTTCGGCT
					AAACTAATCT				AACTATGGCT
					TT				ACACCTTC
	83	S48	CAVSDEDTGF	210	TGTGCTGTGA	211	CASSFSTCSA	114	TGTGCCAGCA	115
			QKLVF		GTGACGAGGA		NYGYTF		GTTTCTCGAC
					CACAGGCTTT				CTGTTCGGCT
					CAGAAACTTG				AACTATGGCT
					TATTT				ACACCTTC
	84	S48	out_of_	N/A	TGCAATACCC	212	CASSFSTCSA	114	TGTGCCAGCA	115
			frame		CAACCAAGGA		NYGYTF		GTTTCTCGAC
					CTCCAGCTTC				CTGTTCGGCT
					TCCTGAAGTA				AACTATGGCT
					CACATCAGCG				ACACCTTC
					GCCACCCTGG
					TTAAAGGCAT
					CAACGGTTTT
					GAGGCTGAAT
					TTAAGAAGAG
					TGAAACCTCC
					TTCCACCTGA
					CGAAACCCTC
					AGCCCATATG
					AGCGACGCGG
					CTGAGTACTT
					CTGTGCTGAG
					TGATCTCGAA
					CCGAACAGCA
					GTGCTTCCAA
					GATAATCTTT
	85	S48	CALSEDRGST	118	TGTGCTCTGA	119	CASSFSTCSA	114	TGTGCCAGCA	115
			LGRLYF		GTGAAGACAG		NYGYTF		GTTTCTCGAC
					AGGCTCAACC				CTGTTCGGCT
					CTGGGGAGGC				AACTATGGCT
					TATACTTT				ACACCTTC
	86	S48	CALQAGGGAN	207	TGTGCTCTGC	208	CASSFSTCSA	114	TGTGCCAGCA	115
			SKLTF		AAGCGGGAGG		NYGYTF		GTTTCTCGAC
					TGGAGCCAAT				CTGTTCGGCT
					AGTAAGCTGA				AACTATGGCT
					CATTT				ACACCTTC
	87	S48	CAVSDLEPNS	152	TGTGCTGTGA	153	CASSFSTCSA	114	TGTGCCAGCA	115
			SASKIIF		GTGATCTCGA		NYGYTF		GTTTCTCGAC
					ACCGAACAGC				CTGTTCGGCT
					AGTGCTTCCA				AACTATGGCT
					AGATAATCTT				ACACCTTC
					T

TABLE 16
			TCRA	TCRB
				SEQ		SEQ		SEQ		SEQ
		Sam-	CDR3	ID	CDR3	ID	CDR3	ID	CDR3	ID
	Row	ple	(aa)	NO	(nt)	NO	(aa)	NO	(nt)	NO
Donor	1	S26	CKLQL	172	TGCAA	173	CASSQ	213	TGTGC	214
19053796			LNLET		ATTGC		GDRGP		CAGCA
			QLSTF		AGCTA		SNTEA		GCCAA
			VPENT		CTCAA		FF		GGGGA
			GGFKT		CCTGG				CAGGG
			IF		AGACT				GGCCG
					CAGCT				TCGAA
					GTCTA				CACTG
					CTTTT				AAGCT
					GTGCC				TTCTT
					TGAAA				T
					ATACT
					GGAGG
					CTTCA
					AAACT
					ATCTT
					T
	2	S26	CREHG	168	TGCCG	169	CASSP	170	TGTGC	171
			DDMRF		TGAAC		LRDNT		CAGCT
					ATGGC		EAFF		CACCA
					GATGA				CTTCG
					CATGC				GGACA
					GCTTT				ACACC
									GAAGC
									TTTCT
									TT
	3	S26	CKLQL	172	TGCAA	173	CASSQ	213	TGTGC	214
			LNLET		ATTGC		GDRGP		CAGCA
			QLSTF		AGCTA		SNTEA		GCCAA
			VPENT		CTCAA		FF		GGGGA
			GGFKT		CCTGG				CAGGG
			IF		AGACT				GGCCG
					CAGCT				TCGAA
					GTCTA				CACTG
					CTTTT				AAGCT
					GTGCC				TTCTT
					TGAAA				T
					ATACT
					GGAGG
					CTTCA
					AAACT
					ATCTT
					T
	4	S26	CKLQL	172	TGCAA	173	CASSP	170	TGTGC	171
			LNLET		ATTGC		LRDNT		CAGCT
			QLSTF		AGCTA		EAFF		CACCA
			VPENT		CTCAA				CTTCG
			GGFKT		CCTGG				GGACA
			IF		AGACT				ACACC
					CAGCT				GAAGC
					GTCTA				TTTCT
					CTTTT				TT
					GTGCC
					TGAAA
					ATACT
					GGAGG
					CTTCA
					AAACT
					ATCTT
					T
	5	S27	CAPEE	215	TGTGC	216	CASSF	114	TGTGC	115
			NYGKN		TCCCG		STCSA		CAGCA
			FVF		AGGAG		NYGYT		GTTTC
					AACTA		F		TCGAC
					TGGTA				CTGTT
					AGAAT				CGGCT
					TTTGT				AACTA
					CTTT				TGGCT
									ACACC
									TTC
	6	S29	CKLQL	172	TGCAA	173	CASSF	114	TGTGC	115
			LNLET		ATTGC		STCSA		CAGCA
			QLSTF		AGCTA		NYGYT		GTTTC
			VPENT		CTCAA		F		TCGAC
			GGFKT		CCTGG				CTGTT
			IF		AGACT				CGGCT
					CAGCT				AACTA
					GTCTA				TGGCT
					CTTTT				ACACC
					GTGCC				TTC
					TGAAA
					ATACT
					GGAGG
					CTTCA
					AAACT
					ATCTT
					T
	7	S30	CAAIG	217	TGTGC	218	CASSF	114	TGTGC	115
			YGENF		AGCAA		STCSA		CAGCA
			VF		TCGGC		NYGYT		GTTTC
					TATGG		F		TCGAC
					TGAGA				CTGTT
					ATTTT				CGGCT
					GTCTT				AACTA
					T				TGGCT
									ACACC
									TTC
	8	S30	CAVSD	152	TGTGC	153	CASSF	114	TGTGC	115
			LEPNS		TGTGA		STCSA		CAGCA
			SASKI		GTGAT		NYGYT		GTTTC
			IF		CTCGA		F		TCGAC
					ACCGA				CTGTT
					ACAGC				CGGCT
					AGTGC				AACTA
					TTCCA				TGGCT
					AGATA				ACACC
					ATCTT				TTC
					T
	9	S32	CKLQL	172	TGCAA	173	CASSF	114	TGTGC	115
			LNLET		ATTGC		STCSA		CAGCA
			QLSTF		AGCTA		NYGYT		GTTTC
			VPENT		CTCAA		F		TCGAC
			GGFKT		CCTGG				CTGTT
			IF		AGACT				CGGCT
					CAGCT				AACTA
					GTCTA				TGGCT
					CTTTT				ACACC
					GTGCC				TTC
					TGAAA
					ATACT
					GGAGG
					CTTCA
					AAACT
					ATCTT
					T
	10	S32	CAMTR	219	TGTGC	220	CASSF	114	TGTGC	115
			SSNTG		AATGA		STCSA		CAGCA
			KLIF		CCCGT		NYGYT		GTTTC
					TCTAG		F		TCGAC
					CAACA				CTGTT
					CAGGC				CGGCT
					AAACT				AACTA
					AATCT				TGGCT
					TT				ACACC
									TTC
	11	S34	CKLQL	172	TGCAA	173	CASSQ	213	TGTGC	214
			LNLET		ATTGC		GDRGP		CAGCA
			QLSTF		AGCTA		SNTEA		GCCAA
			VPENT		CTCAA		FF		GGGGA
			GGFKT		CCTGG				CAGGG
			IF		AGACT				GGCCG
					CAGCT				TCGAA
					GTCTA				CACTG
					CTTTT				AAGCT
					GTGCC				TTCTT
					TGAAA				T
					ATACT
					GGAGG
					CTTCA
					AAACT
					ATCTT
					T
	12	S34	CAMTR	219	TGTGC	220	CASSR	221	TGTGC	222
			SSNTG		AATGA		DRVGQ		CAGCA
			KLIF		CCCGT		YF		GCCGG
					TCTAG				GACAG
					CAACA				GGTCG
					CAGGC				GGCAG
					AAACT				TACTT
					AATCT				C
					TT
	13	S35	CAMTR	219	TGTGC	220	CASSR	221	TGTGC	222
			SSNTG		AATGA		DRVGQ		CAGCA
			KLIF		CCCGT		YF		GCCGG
					TCTAG				GACAG
					CAACA				GGTCG
					CAGGC				GGCAG
					AAACT				TACTT
					AATCT				C
					TT
	14	S35	CALDM	223	TGTGC	224	CASSR	221	TGTGC	222
			NYGGA		TCTAG		DRVGQ		CAGCA
			TNKLI		ACATG		YF		GCCGG
			F		AATTA				GACAG
					TGGTG				GGTCG
					GTGCT				GGCAG
					ACAAA				TACTT
					CAAGC				C
					TCATC
					TTT
	15	S35	CKLQL	172	TGCAA	173	CASSQ	213	TGTGC	214
			LNLET		ATTGC		GDRGP		CAGCA
			QLSTF		AGCTA		SNTEA		GCCAA
			VPENT		CTCAA		FF		GGGGA
			GGFKT		CCTGG				CAGGG
			IF		AGACT				GGCCG
					CAGCT				TCGAA
					GTCTA				CACTG
					CTTTT				AAGCT
					GTGCC				TTCTT
					TGAAA				T
					ATACT
					GGAGG
					CTTCA
					AAACT
					ATCTT
					T
	16	S35	CAMTR	219	TGTGC	220	CASSR	221	TGTGC	222
			SSNTG		AATGA		DRVGQ		CAGCA
			KLIF		CCCGT		YF		GCCGG
					TCTAG				GACAG
					CAACA				GGTCG
					CAGGC				GGCAG
					AAACT				TACTT
					AATCT				C
					TT
	17	S3 5	CAMTR	219	TGTGC	220	CASSR	221	TGTGC	222
			SSNTG		AATGA		DRVGQ		CAGCA
			KLIF		CCCGT		YF		GCCGG
					TCTAG				GACAG
					CAACA				GGTCG
					CAGGC				GGCAG
					AAACT				TACTT
					AATCT				C
					TT

TABLE 17
			TCRA	TCRB
				SEQ		SEQ		SEQ		SEQ
			CDR3	ID	CDR3	ID	CDR3	ID	CDR3	ID
	Row	Well	(aa)	NO	(nt)	NO	(aa)	NO	(nt)	NO
Donor	1	B1	CILDN	248	TGCAT	249	CASSL	250	TGTGC	251
2001476			NNDMR		CCTTG		APGAT		CAGCA
			F		ACAAT		NEKLF		GCTTA
					AACAA		F		GCGCC
					TGACA				GGGTG
					TGCGC				CAACT
					TTT				AATGA
									AAAAC
									TGTTT
									TTT
	2	C1	CILDN	248	TGCAT	249	CASSL	250	TGTGC	251
			NNDMR		CCTTG		APGAT		CAGCA
			F		ACAAT		NEKLF		GCTTA
					AACAA		F		GCGCC
					TGACA				GGGTG
					TGCGC				CAACT
					TTT				AATGA
									AAAAC
									TGTTT
									TTT
	3	D1	CAVSD	152	TGTGC	153	CATSR	252	TGTGC	253
			LEPNS		TGTGA		DLPLA		CACCA
			SASKI		GTGAT		GGRGE		GCAGA
			IF		CTCGA		QFF		GATCT
					ACCGA				CCCGC
					ACAGC				TAGCG
					AGTGC				GGGGG
					TTCCA				GCGAG
					AGATA				GTGAG
					ATCTT				CAGTT
					T				CTTC
	4	E1	CAVSD	152	TGTGC	153	CASSF	114	TGTGC	115
			LEPNS		TGTGA		STCSA		CAGCA
			SASKI		GTGAT		NYGYT		GTTTC
			IF		CTCGA		F		TCGAC
					ACCGA				CTGTT
					ACAGC				CGGCT
					AGTGC				AACTA
					TTCCA				TGGCT
					AGATA				ACACC
					ATCTT				TTC
					T
	5	F1	CLVVY	254	TGCCT	255	CASSH	256	TGCGC	257
			DYKLS		CGTGG		LTGLA		CAGCA
			F		TCTAC		EAFF		GCCAT
					GACTA				CTGAC
					CAAGC				AGGGT
					TCAGC				TGGCT
					TTT				GAAGC
									TTTCT
									TT
	6	H1	CLVVY	254	TGCCT	255	CASSF	114	TGTGC	115
			DYKLS		CGTGG		STCSA		CAGCA
			F		TCTAC		NYGYT		GTTTC
					GACTA		F		TCGAC
					CAAGC				CTGTT
					TCAGC				CGGCT
					TTT				AACTA
									TGGCT
									ACACC
									TTC
	7	C2	CILDN	248	TGCAT	249	CASSL	258	TGCGC	259
			NNDMR		CCTTG		LGNSP		CAGCA
			F		ACAAT		LHF		GTCTG
					AACAA				CTGGG
					TGACA				TAATT
					TGCGC				CACCC
					TTT				CTCCA
									CTTT
	8	D2	out_		TGTTC	260	CASSG	261	TGCGC	262
			of_		ATCAG		LAGAY		CAGCA
			frame		AGACT		NEQFF		GCGGG
					CACAG				CTAGC
					CCCAG				GGGGG
					TGATT				CCTAC
					CAGCC				AATGA
					ACCTA				GCAGT
					CCTCT				TCTTC
					GTGCA
					ATGAC
					CTGCC
					ACTGA
					CCTTC
					AGGGA
					GCCCA
					GAAGC
					TGGTA
					TTT
	9	E2	CAATH	263	TGTGC	264	CASSL	265	TGTGC	266
			SKSGY		AGCAA		WVMNT		CAGCA
			ALNF		CCCAC		EAFF		GCTTG
					TCAAA				TGGGT
					TTCCG				TATGA
					GGTAT				ACACT
					GCACT				GAAGC
					CAACT				TTTCT
					TC				TT
	10	E3	CELDN	248	TGCAT	249	CASSL	250	TGTGC	251
			NNDMR		CCTTG		APGAT		CAGCA
			F		ACAAT		NEKLF		GCTTA
					AACAA		F		GCGCC
					TGACA				GGGTG
					TGCGC				CAACT
					TTT				AATGA
									AAAAC
									TGTTT
									TTT
	11	D6	CAVYG	267	TGTGC	268	CASSI	269	TGTGC	270
			GATKK		TGTTT		GEAFF		CAGCA
			LIF		ATGGT				GTATT
					GGTGC				GGGGA
					TACAA				AGCTT
					ACAAG				TCTTT
					CTCAT
					CTTT
	12	H6	CAVYG	267	TGTGC	268	CASTP	271	TGTGC	272
			GATKK		TGTTT		GTGAY		CAGCA
			LIF		ATGGT		EQYF		CCCCC
					GGTGC				GGGAC
					TACAA				AGGGG
					ACAAG				CGTAC
					CTCAT				GAGCA
					CTTT				GTACT
									TC
	13	E7	CAVSD	152	TGTGC	153	CASSF	114	TGTGC	115
			LEPNS		TGTGA		STCSA		CAGCA
			SASKI		GTGAT		NYGYT		GTTTC
			IF		CTCGA		F		TCGAC
					ACCGA				CTGTT
					ACAGC				CGGCT
					AGTGC				AACTA
					TTCCA				TGGCT
					AGATA				ACACC
					ATCTT				TTC
					T
	14	F7	CAVRA	273	TGTGC	274	CASND	275	TGTGC	276
			LVPGA		TGTGA		YSSPL		CAGCA
			GSYQL		GAGCC		HF		ACGAC
			TF		CTCGT				TATAG
					CCCTG				TTCAC
					GGGCT				CCCTC
					GGGAG				CACTT
					TTACC				T
					AACTC
					ACTTT
					C
	15	B8	CAVNR	277	TGTGC	278	CASSY	279	TGTGC	280
			GGGNK		CGTGA		GGAYE		CAGCA
			LTF		ACCGG		QYF		GTTAT
					GGAGG				GGGGG
					AGGAA				AGCCT
					ACAAA				ACGAG
					CTCAC				CAGTA
					CTTT				CTTC
	16	D8	CAVSD	152	TGTGC	153	CASSF	114	TGTGC	115
			LEPNS		TGTGA		STCSA		CAGCA
			SASKI		GTGAT		NYGYT		GTTTC
			IF		CTCGA		F		TCGAC
					ACCGA				CTGTT
					ACAGC				CGGCT
					AGTGC				AACTA
					TTCCA				TGGCT
					AGATA				ACACC
					ATCTT				TTC
					T
	17	F8	CAVSD	152	TGTGC	153	CASSF	114	TGTGC	115
			LEPNS		TGTGA		STCSA		CAGCA
			SASKI		GTGAT		NYGYT		GTTTC
			IF		CTCGA		F		TCGAC
					ACCGA				CTGTT
					ACAGC				CGGCT
					AGTGC				AACTA
					TTCCA				TGGCT
					AGATA				ACACC
					ATCTT				TTC
					T
	18	E9	CISWI	281	TGCAT	282	CASSP	283	TGTGC	284
			PSLET		ATCAT		VQGVY		CAGCA
			QPPPL		GGATT		NEQFF		GCCCA
			RSQLI		CCCAG				GTCCA
			F		CCTGG				GGGGG
					AGACT				TTTAC
					CAGCC				AATGA
					ACCCC				GCAGT
					CCTTG				TCTTC
					AGGTC
					GCAAC
					TCATC
					TTT
	19	F9	CATPR	285	TGTGC	286	CASSL	287	TGTGC	288
			YF		AACCC		AGETQ		CAGCA
					CGCGC		YF		GCCTC
					TATTT				GCGGG
					T				AGAGA
									CCCAG
									TACTT
									C
	20	B10	CILDN	248	TGCAT	249	CASSL	250	TGTGC	251
			NNDMR		CCTTG		APGAT		CAGCA
			F		ACAAT		NEKLF		GCTTA
					AACAA		F		GCGCC
					TGACA				GGGTG
					TGCGC				CAACT
					TTT				AATGA
									AAAAC
									TGTTT
									TTT
	21	C10	CILDN	248	TGCAT	249	CASSL	250	TGTGC	251
			NNDMR		CCTTG		APGAT		CAGCA
			F		ACAAT		NEKLF		GCTTA
					AACAA		F		GCGCC
					TGACA				GGGTG
					TGCGC				CAACT
					TTT				AATGA
									AAAAC
									TGTTT
									TTT

TABLE 18
			TCRA	TCRB
				SEQ		SEQ		SEQ		SEQ
			CDR3	ID	CDR3	ID	CDR3	ID	CDR3	ID
	Row	Well	(aa)	NO	(nt)	NO	(aa)	NO	(nt)	NO
Donor	1	E3	CILDN	248	TGCAT	249	CASSF	114	TGTGC	115
2001476			NNDMR		CCTTG		STCSA		CAGCA
			F		ACAAT		NYGYT		GTTTC
					AACAA		F		TCGAC
					TGACA				CTGTT
					TGCGC				CGGCT
					TTT				AACTA
									TGGCT
									ACACC
									TTC
	2	B4	CAASL	289	TGTGC	290	CASSQ	291	TGTGC	292
			IGKJL		AGCAA		LQSSY		CAGCA
			TF		GCCTT		NEQFF		GCCAA
					ATAGG				TTACA
					GAAAC				GAGCT
					TGACA				CCTAC
					TTT				AATGA
									GCAGT
									TCTTC
	3	C4	CAASL	289	TGTGC	290	CASSQ	291	TGTGC	292
			IGKLT		AGCAA		LQSSY		CAGCA
			F		GCCTT		NEQFF		GCCAA
					ATAGG				TTACA
					GAAAC				GAGCT
					TGACA				CCTAC
					TTT				AATGA
									GCAGT
									TCTTC
	4	E4	CAASL	289	TGTGC	290	CASSF	114	TGTGC	115
			IGKLT		AGCAA		STCSA		CAGCA
			F		GCCTT		NYGYT		GTTTC
					ATAGG		F		TCGAC
					GAAAC				CTGTT
					TGACA				CGGCT
					TTT				AACTA
									TGGCT
									ACACC
									TTC
	5	F4	CAASL	289	TGTGC	290	CASSF	114	TGTGC	115
			IGKLT		AGCAA		STCSA		CAGCA
			F		GCCTT		NYGYT		GTTTC
					ATAGG		F		TCGAC
					GAAAC				CTGTT
					TGACA				CGGCT
					TTT				AACTA
									TGGCT
									ACACC
									TTC

Table 19, Table 20, Table 21, Table 22 and Table 23 provide the CDR1 and CDR2 amino acid (aa) sequences and the CDR1 and CDR2 nucleotide (nt) sequences and SEQ ID NOs for select mutant-mimic pair SEQ ID NO: 9 and SEQ ID NO: 45 tetramer positive TCRs identified in Table 9, the mutant-mimic pair SEQ ID NO: 13 and SEQ ID NO: 59 tetramer positive TCRs identified in Table 10, the mutant-mimic pair SEQ ID NO: 18 and SEQ ID NO: 68 tetramer positive TCRs identified in Table 11, the mutant-mimic pair SEQ ID NO: 3 and SEQ ID NO: 32 tetramer positive TCRs identified in Table 12, and the mutant-mimic pair SEQ ID NO: 23 and SEQ ID NO: 78 tetramer positive TCRs identified in Table 13, respectively. Each row in Table 19, Table 20, Table 21, Table 22, and Table 23 corresponds to the matching row in Table 9, Table 10, Table 11, Table 12 and Table 13, respectively, e.g., the V(J) or V(D)J genes in row 1 of Table 9 correspond to the CDR1 and CDR2 sequences in row 1 of Table 19, and so on.

TABLE 19
			TCRA	TCRB
			CDR	SEQ	CDR	SEQ	CDR	SEQ	CDR	SEQ	CDR	SEQ	CDR	SEQ	CDR	SEQ	CDR	SEQ
		Sam-	1	ID	1	ID	2	ID	2	ID	1	ID	1	ID	2	ID	2	ID
	Row	ple	(aa)	NO	(nt)	NO	(aa)	NO	(nt)	NO	(aa)	NO	(nt)	NO	(aa)	NO	(nt)	NO
Donor	1	S17	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—
19054445
	2	S19	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—
	3	S20	—	—	—	—	—	—	—	—	SGH	293	TCA	294	FNN	295	TTT	296
											NS		GGC		NVP		AAC
													CAC				AAC
													AAC				AAC
													TCC				GTT
																	CCG
	4	S21	—	—	—	—	—	—	—	—	SGH	293	TCA	294	FNN	295	TTT	296
											NS		GGC		NVP		AAC
													CAC				AAC
													AAC				AAC
													TCC				GTT
																	CCG
	5	S22	—	—	—	—	—	—	—	—	SGH	293	TCA	294	FNN	295	TTT	296
											NS		GGC		NVP		AAC
													CAC				AAC
													AAC				AAC
																	GTT
																	CCG
	6	S24	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—
	7	S24	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—
	8	S25	—	—	—	—	—	—	—	—	SGH	293	TCA	294	FNN	295	TTT	296
											NS		GGC		NVP		AAC
													CAC				AAC
													AAC				AAC
													TCC				GTT
																	CCG
Donor	9	S39	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—
19063796
	10	S41	—	—	—	—	—	—	—	—	DFQ	297	GAC	298	SNE	299	TCC	300
											ATT		TTTC		GSK		AAT
													AGG		A		GAG
													CCA				GGC
													CAA				TCC
													CT				AAG
																	GCC
	11	S41	YSG	301	TAT	302	HISR	303	CAC	304	DFQ	297	GAC	298	SNE	299	TCC	300
			SPE		TCT				ATC		ATT		TTTC		GSK		AAT
					GGG				TCT				AGG		A		GAG
					AGT				AGA				CCA				GGC
					CCT								CAA				TCC
					GAA								CT				AAG
																	GCC
	12	S41	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—
	13	S41	—	—	—	—	—	—	—	—	SGH	305	TCT	306	FQD	307	TTTC	308
											AT		GGC		ESV		AGG
													CAT				ATG
													GCT				AGA
													ACC				GTG
																	TA
	14	S42	DSSS	309	GAC	310	IFSN	311	ATT	312	—	—	—	—	—	—	—	—
			TY		AGC		MDM		TTTT
					TCC				CAA
					TCC				ATA
					ACC				TGG
					TAC				ACA
									TG
	15	S42	—	—	—	—	—	—	—	—	SGH	293	TCA	294	FNN	295	TTT	296
											NS		GGC		NVP		AAC
													CAC				AAC
													AAC				AAC
													TCC				GTT
																	CCG
	16	S44	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—
	17	S44	—	—	—	—	—	—	—	—	SGH	305	TCT	306	FQD	307	TTTC	308
											AT		GGC		ESV		AGG
													CAT				ATG
													GCT				AGA
													ACC				GTG
																	TA
	18	S45	DSSS	309	GAC	310	IFSN	311	ATT	312	SGH	305	TCT	306	FQD	307	TTTC	308
			TY		AGC		MDM		TTTT		AT		GGC		ESV		AGG
					TCC				CAA				CAT				ATG
					TCC				ATA				GCT				AGA
					ACC				TGG				ACC				GTG
					TAC				ACA								TA
									TG
	19	S45	DSSS	309	GAC	310	IFSN	311	ATT	312	SGH	305	TCT	306	FQD	307	TTTC	308
			TY		AGC		MDM		TTTT		AT		GGC		ESV		AGG
					TCC				CAA				CAT				ATG
					TCC				ATA				GCT				AGA
					ACC				TGG				ACC				GTG
					TAC				ACA								TA
									TG
	20	S45	DSSS	309	GAC	310	IFSN	311	ATT	312	SGH	305	TCT	306	FQD	307	TTTC	308
			TY		AGC		MDM		TTTT		AT		GGC		ESV		AGG
					TCC				CAA				CAT				ATG
					TCC				ATA				GCT				AGA
					ACC				TGG				ACC				GTG
					TAC				ACA								TA
									TG
	21	S45	DSSS	309	GAC	310	IFSN	311	ATT	312	SGH	313	TCT	314	YYE	315	TAT	316
			TY		AGC		MDM		TTTT		DT		GGG		EEE		TAT
					TCC				CAA				CAT				GAG
					TCC				ATA				GAC				GAG
					ACC				TGG				ACT				GAA
					TAC				ACA								GAG
									TG
	22	S46	—	—	—	—	—	—	—	—	LNH	317	TTG	318	YYD	319	TAC	320
											NV		AAC		KDF		TAT
													CAT				GAC
													AAC				AAA
													GTC				GAT
																	TTT
	23	S48	—	—	—	—	—	—	—	—		—		—		—		—
	24	S50	—	—	—	—	—	—	—	—		—		—		—		—

TABLE 20
			TCRA
				SEQ		SEQ		SEQ		SEQ
			CDR	ID	CDR	ID	CDR	ID	CDR	ID
	Row	Sample	1 (aa)	NO	1 (nt)	NO	2 (aa)	NO	2 (nt)	NO
Donor	1	S27	SSNF	321	TCC	322	MTL	323	ATG	324
19054445			YA		AGC		NGD		ACT
					AAT		E		TTA
					TTTT				AAT
					ATG				GGG
					CC				GAT
									GAA
	2	S28	—	—	—	—	—	—	—	—
	3	S29	—	—	—	—	—	—	—	—
	4	S29	—	—	—	—	—	—	—	—
	5	S29	SSNF	321	TCC	322	MTL	323	ATG	324
			YA		AGC		NGD		ACT
					AAT		E		TTA
					TTTT				AAT
					ATG				GGG
					CC				GAT
									GAA
	6	S29	SSNF	321	TCC	322	MTL	323	ATG	324
			YA		AGC		NGD		ACT
					AAT		E		TTA
					TTTT				AAT
					ATG				GGG
					CC				GAT
									GAA
	7	S30	—	—	—	—	—	—	—	—
	8	S30	—	—	—	—	—	—	—	—
	9	S30	SSVP	333	TCG	334	YTS	335	TAC	336
			PY		TCT		AAT		ACA
					GTT		LV		TCA
					CCA				GCG
					CCA				GCC
					TAT				ACC
									CTG
									GTT
	10	S31	—	—	—	—	—	—	—	—
	11	S31	—	—	—	—	—	—	—	—
	12	S32	—	—	—	—	—	—	—	—
	13	S32	—	—	—	—	—	—	—	—
	14	S32	ATG	337	GCC	338	ATK	399	GCC	340
			YPS		ACA		ADD		ACA
					GGA		K		AAG
					TAC				GCT
					CCT				GAT
					TCC				GAC
									AAG
	15	S32	—	—	—	—	—	—	—	—
	16	S32	—	—	—	—	—	—	—	—
	17	S32	ATG	337	GCC	338	ATK	399	GCC	340
			YPS		ACA		ADD		ACG
					GGA		K		AAG
					TAC				GCT
					CCT				GAT
					TCC				GAC
									AAG
	18	S33	DSA	341	GAC	342	IRSN	343	ATT	344
			SNY		AGT		VGE		CGT
					GCC				TCA
					TCA				AAT
					AAC				GTG
					TAC				GGC
									GAA
	19	S33	—	—	—	—	—	—	—	—
	20	S33	SSVP	333	TCG	334	YTS	335	TAC	336
			PY		TCT		AAT		ACA
					GTT		LV		TCA
					CCA				GCG
					CCA				GCC
					TAT				ACC
									CTG
									GTT
	21	S33	—	—	—	—	—	—	—	—
	22	S33	SSVP	333	TCG	334	YTS	349	TAC	350
			PY		TCT		AAT		ACA
					GTT		LA		TCA
					CCA				GCG
					CCA				GCC
					TAT				ACC
									CTG
									GCT
	23	S33	—	—	—	—	—	—	—	—
	24	S33	—	—	—	—	—	—	—	—
	25	S34	NIAT	351	AAC	352	GYK	353	GGA	354
			NDY		ATT		TK		TAC
					GCT				AAG
					ACA				ACA
					AAT				AAA
					GAT
					TAT
	26	S33	—	—	—	—	—	—	—	—
	27	S33	—	—	—	—	—	—	—	—
	28	S34	NIAT	351	AAC	352	GYK	353	GGA	354
			NDY		ATT		TK		TAC
					GCT				AAG
					ACA				ACA
					AAT				AAA
					GAT
					TAT
	29	S33	—	—	—	—	—	—	—	—
	30	S33	—	—	—	—	—	—	—	—
	31	S33	—	—	—	—	—	—	—	—
	32	S33	—	—	—	—	—	—	—	—
	33	S33	—	—	—	—	—	—	—	—
	34	S33	—	—	—	—	—	—	—	—
Donor	35	S14	—	—	—	—	—	—	—	—
19053796	36	S14	—	—	—	—	—	—	—	—
	37	S14	—	—	—	—	—	—	—	—
	38	S15	AQK	366	ACA	367	QGS	368	CAG	369
			VTQ		AGT				GGT
			AQS		TGG				TCT
			SVS		TGG
			MPV		TCA
			RKA		TAT
			VTL		TAT
			NCL
			YE
	39	S15	AQK	366	ACA	367	QGS	368	CAG	369
			VTQ		AGT				GGT
			AQS		TGG				TCT
			SVS		TGG
			MPV		TCA
			RKA		TAT
			VTL		TAT
			NCL
			YE
	40	S15	INAE	376	TCG	334	YTS	335	TAC	336
			YQIG		TCT		AAT		ACA
			SHV		GTT		LV		TCA
			SVSE		CCA				GCG
			GAL		CCA				GCC
			VLL		TAT				ACC
			RCN						CTG
			YS						GTT
	41	S15	—	—	—	—	—	—	—	—
	42	S16	KNQ	377	GTG	378	MTF	379	ATG	380
			VEQ		AGC		SENT		ACT
			SPQS		CCC				TTC
			LIILE		TTC				AGT
			GKN		AGC				GAG
			CTL		AAC				AAC
			QCN						ACA
			YT
	43	S16	KNQ	377	GTG	378	MTF	379	ATG	380
			VEQ		AGC		SENT		ACT
			SPQS		CCC				TTC
			LIILE		TTC				AGT
			GKN		AGC				GAG
			CTL		AAC				AAC
			QCN						ACA
			YT
	44	S16	KNQ	377	GTG	378	MTF	379	ATG	380
			VEQ		AGC		SENT		ACT
			SPQS		CCC				TTC
			LIILE		TTC				AGT
			GKN		AGC				GAG
			CTL		AAC				AAC
			QCN						ACA
			YT
	45	S16	KNQ	377	GTG	378	MTF	379	ATG	380
			VEQ		AGC		SENT		ACT
			SPQS		CCC				TTC
			LIILE		TTC				AGT
			GKN		AGC				GAG
			CTL		AAC				AAC
			QCN						ACA
			YT
	46	S16	KNQ	377	GTG	378	MTF	379	ATG	380
			VEQ		AGC		SENT		ACT
			SPQS		CCC				TTC
			LIILE		TTC				AGT
			GKN		AGC				GAG
			CTL		AAC				AAC
			QCN						ACA
			YT
	47	S17	—	—	—	—	—	—	—	—
	48	S17	—	—	—	—	—	—	—	—
	49	S17	—	—	—	—	—	—	—	—
	50	S18	—	—	—	—	—	—	—	—
	51	S18	—	—	—	—	—	—	—	—
	52	S18	—	—	—	—	—	—	—	—
	53	S18	GQQ	389	GTG	378	MTF	379	ATG	380
			VMQ		AGC		SENT		ACT
			IPQY		CCC				TTC
			QHV		TTC				AGT
			QEG		AGC				GAG
			EDFT		AAC				AAC
			TYC						ACA
			NSS
	54	S18	—	—	—	—	—	—	—	—
	55	S19	INAE	393	TCG	334	YTS	335	TAC	336
			YRC		TCT		AAT		ACA
			GSH		GTT		LV		TCA
			VSV		CCA				GCG
			SEG		CCA				GCC
			ALV		TAT				ACC
			LLR						CTG
			CNY						GTT
			S
	56	S19	INAE	376	TCG	334	YTS	335	TAC	336
			YQIG		TCT		AAT		ACA
			SHV		GTT		LV		TCA
			SVSE		CCA				GCG
			GAL		CCA				GCC
			VLL		TAT				ACC
			RCN						CTG
			YS						GTT
	57	S20	—	—	—	—	—	—	—	—
	58	S20	—	—	—	—	—	—	—	—
	59	S20	—	—	—	—	—	—	—	—
	60	S23	—	—	—	—	—	—	—	—
Donor	61	S38	TISG	394	ACC	395	GLK	396	GGT	397
			NEY		ATC		NN		CTA
					AGT				AAA
					GGA				AAC
					AAT				AAT
					GAG
					TAT
	62	S38	ATG	337	GCC	338	ATK	339	GCC	340
			YPS		ACA		ADD		ACG
					GGA		K		AAG
					TAC				GCT
					CCT				GAT
					TCC				GAC
									AAG
	63	S38	—	—	—	—	—	—	—	—
	64	S38	—	—	—	—	—	—	—	—
	65	S38	—	—	—	—	—	—	—	—
	66	S39	—	—	—	—	—	—	—	—
	67	S40	—	—	—	—	—	—	—	—
	68	S42	—	—	—	—	—	—	—	—
	69	S42	—	—	—	—	—	—	—	—
	70	S43	—	—	—	—	—	—	—	—
	71	S44	—	—	—	—	—	—	—	—
	72	S45	—	—	—	—	—	—	—	—
	73	S46	SSVP	333	TCG	334	YTS	335	TAC	336
			PY		TCT		AAT		ACA
					GTT		LV		TCA
					CCA				GCG
					CCA				GCC
					TAT				ACC
									CTG
									GTT
	74	S46	NIAT	351	AAC	352	GYK	353	GGA	354
			NDY		ATT		TK		TAC
					GCT				AAG
					ACA				ACA
					AAT				AAA
					GAT
					TAT
	75	S46	—	—	—	—	—	—	—	—
	76	S46	—	—	—	—	—	—	—	—
	77	S46	ATG	337	GCC	338	ATK	339	GCC	340
			YPS		ACA		ADD		ACG
					GGA		K		AAG
					TAC				GCT
					CCT				GAT
					TCC				GAC
									AAG
	78	S46	ATG	337	GCC	338	ATK	339	GCC	340
			YPS		ACA		ADD		ACG
					GGA		K		AAG
					TAC				GCT
					CCT				GAT
					TCC				GAC
									AAG
	79	S46	—	—	—	—	—	—	—	—
	80	S46	ATG	337	GCC	338	ATK	339	GCC	340
			YPS		ACA		ADD		ACG
					GGA		K		AAG
					TAC				GCT
					CCT				GAT
					TCC				GAC
									AAG
	81	S47	ATG	337	GCC	338	ATK	339	GCC	340
			YPS		ACA		ADD		ACG
					GGA		K		AAG
					TAC				GCT
					CCT				GAT
					TCC				GAC
									AAG
	82	S47	ATG	337	GCC	338	ATK	339	GCC	340
			YPS		ACA		ADD		ACG
					GGA		K		AAG
					TAC				GCT
					CCT				GAT
					TCC				GAC
									AAG
	83	S48	—	—	—	—	—	—	—	—
	84	S48	—	—	—	—	—	—	—	—
	85	S48	—	—	—	—	—	—	—	—
	86	S48	ATG	337	GCC	338	ATK	339	GCC	340
			YPS		ACA		ADD		ACG
					GGA		K		AAG
					TAC				GCT
					CCT				GAT
					TCC				GAC
									AAG
	87	S48	—	—	—	—	—	—	—	—
			TCRB
				SEQ		SEQ		SEQ		SEQ
			CDR	ID	CDR	ID	CDR	ID	CDR	ID
	Row	Sample	1 (aa)	NO	1 (nt)	NO	2 (aa)	NO	2 (nt)	NO
Donor	1	S27	SNH	325	TCT	326	FYN	327	TTTT	328
19054445			LY		AAT		NEI		ATA
					CAC				ATA
					TTA				ATG
					TAC				AAA
									TC
	2	S28
	3	S29	SGH	329	TCG	330	FQN	331	TTC	332
			VS		GGT		EAQ		CAG
					CAT				AAT
					GTA				GAA
					TCC				GCT
									CAA
	4	S29	—	—	—	—	—	—	—	—
	5	S29	—	—	—	—	—	—	—	—
	6	S29	—	—	—	—	—	—	—	—
	7	S30	—	—	—	—	—	—	—	—
	8	S30	—	—	—	—	—	—	—	—
	9	S30	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	10	S31	—	—	—	—	—	—	—	—
	11	S31	—	—	—	—	—	—	—	—
	12	S32	—	—	—	—	—	—	—	—
	13	S32	—	—	—	—	—	—	—	—
	14	S32	—	—	—	—	—	—	—	—
	15	S32	—	—	—	—	—	—	—	—
	16	S32	—	—	—	—	—	—	—	—
	17	S32	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	18	S33	MNH	345	ATG	346	SVG	347	TCA	348
			NY		AAC		AGI		GGT
					CAT				GGT
					AAC				GCT
					TAC				GGT
									ATC
	19	S33	—	—	—	—	—	—	—	—
	20	S33	—	—	—	—	—	—	—	—
	21	S33	—	—	—	—	—	—	—	—
	22	S33	—	—	—	—	—	—	—	—
	23	S33	—	—	—	—	—	—	—	—
	24	S33	—	—	—	—	—	—	—	—
	25	S34	—	—	—	—	—	—	—	—
	26	S34	—	—	—	—	—	—	—	—
	27	S34	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	28	S34	MNH	355	ATG	356	SVG	357	TCA	358
			EY		AAC		EGT		GTT
					CAT				GGT
					GAA				GAG
					TAC				GGT
									ACA
	29	S34	—	—	—	—	—	—	—	—
	30	S36	MNH	355	ATG	359	SMN	360	TCA	361
			EY		AAC		VEV		ATG
					CAT				AAT
					GAG				GTT
					TAT				GAG
									GTG
	31	S38	SGH	362	TCA	363	FNN	295	TTT	296
			DY		GGA		NVP		AAC
					CAC				AAC
					GAC				AAC
					TAC				GTT
									CCG
	32	S38	SGH	362	TCA	363	FNN	295	TTT	296
			DY		GGA		NVP		AAC
					CAC				AAC
					GAC				AAC
					TAC				GTT
									CCG
	33	S38	SGH	362	TCA	363	FNN	295	TTT	296
			DY		GGA		NVP		AAC
					CAC				AAC
					GAC				AAC
					TAC				GTT
									CCG
	34	S38	—	—	—	—	—	—	—	—
Donor	35	S14	SEH	364	TCT	365	FQN	331	TTC	332
19053796			NR		GAA		EAQ		CAG
					CAC				AAT
					AAC				GAA
					CGC				GCT
									CAA
	36	S14	—	—	—	—	—	—	—	—
	37	S14	—	—	—	—	—	—	—	—
	38	S15	SGH	305	TCT	306	FQN	370	TTTC	371
			AT		GGC		NGV		AGA
					CAT				ATA
					GCT				ACG
					ACC				GTG
									TA
	39	S15	SGH	372	TCT	373	YFSE	374	TAC	375
			RS		GGG		TQ		TTC
					CAT				AGT
					AGG				GAG
					AGT				ACA
									CAG
	40	S15	—	—	—	—	—	—	—	—
	41	S15	SGH	305	TCT	306	FQN	370	TTTC	371
			AT		GGC		NGV		AGA
					CAT				ATA
					GCT				ACG
					ACC				GTG
									TA
	42	S16	—	—	—	—	—	—	—	—
	43	S16	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	44	S16	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	45	S16	—	—	—	—	—	—	—	—
	46	S16	SQV	381	AGC	382	AVQ	383	GCA	384
			TM		CAA		GSE		AAT
					GTC		A		CAG
					ACC				GGC
					ATG				TCT
									GAG
									GCC
	47	S17	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	48	S17	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	49	S17	—	—	—	—	—	—	—	—
	50	S18	WSH	385	TGG	386	SAA	387	TCA	388
			SY		AGC		ADI		GCA
					CAC				GCT
					AGC				GCT
					TAT				GAT
									ATT
	51	S18	WSH	385	TGG	386	SAA	387	TCA	388
			SY		AGC		ADI		GCA
					CAC				GCT
					AGC				GCT
					TAT				GAT
									ATT
	52	S18	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	53	S18	MNH	355	ATG	356	SVG	347	TCA	348
			EY		AAC		AGI		GTT
					CAT				GGT
					GAA				GCT
					TAC				GGT
									ATC
	54	S18	—	—	—	—	—	—	—	—
	55	S19	—	—	—	—	—	—	—	—
	56	S19	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	57	S20	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	58	S20	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	59	S22	SEH	364	TCT	365	FQN	331	TTC	332
			NR		GAA		EAQ		CAG
					CAC				AAT
					AAC				GAA
					CGC				GCT
									CAA
	60	S23	—	—	—	—	—	—	—	—
Donor	61	S38	PRH	398	CCT	399	FYE	400	TTTT	401
			DT		AGA		KMQ		ATG
					CAC				AAA
					GAC				AGA
					ACT				TGC
									AG
	62	S38	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	63	S38	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	64	S38	—	—	—	—	—	—	—	—
	65	S38	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	66	S39	SGH	305	TCT	306	FQN	370	TTTC	371
			AT		GGC		NGV		AGA
					CAT				ATA
					GCT				ACG
					ACC				GTG
									TA
	67	S40	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	68	S42	—	—	—	—	—	—	—	—
	69	S42	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	70	S43	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	71	S44	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	72	S45	—	—	—	—	—	—	—	—
	73	S46	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	74	S46	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	75	S46	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	76	S46	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	77	S46	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	78	S46	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	79	S46	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	80	S46	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	81	S47	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	82	S47	SGH	293	TCA	294	FNN	402	TTT	403
			NS		GGC		NVS		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									TCG
	83	S48	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	84	S48	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	85	S48	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	86	S48	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	87	S48	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
indicates data missing or illegible when filed

TABLE 21
			TCRA
				SEQ		SEQ		SEQ		SEQ
			CDR	ID	CDR	ID	CDR	ID	CDR	ID
	Row	Sample	1 (aa)	NO	1 (nt)	NO	2 (aa)	NO	2 (nt)	NO
Donor	1	S26	—	—	—	—	—	—	—	—
	2	S26	—	—	—	—	—	—	—	—
	3	S26	—	—	—	—	—	—	—	—
	4	S26	—	—	—	—	—	—	—	—
	5	S27	—	—	—	—	—	—	—	—
	6	S29	—	—	—	—	—	—	—	—
	7	S30	—	—	—	—	—	—	—	—
	8	S30	—	—	—	—	—	—	—	—
	9	S32	—	—	—	—	—	—	—	—
	10	S32	—	—	—	—	—	—	—	—
	11	S34	—	—	—	—	—	—	—	—
	12	S34	NSA	408	AAC	409	TYSS	410	ACA	411
			FQY		AGT		GN		TAC
					GCT				TCC
					TTTC				AGT
					AAT				GGT
					AC				AAC
	13	S35	—	—	—	—	—	—	—	—
	14	S35	—	—	—	—	—	—	—	—
	15	S35	—	—	—	—	—	—	—	—
	16	S35	—	—	—	—	—	—	—	—
	17	S35	—	—	—	—	—	—	—	—
			TCRB
				SEQ		SEQ		SEQ		SEQ
			CDR	ID	CDR	ID	CDR	ID	CDR	ID
	Row	Sample	1 (aa)	NO	1 (nt)	NO	2 (aa)	NO	2 (nt)	NO
Donor	1	S26	—	—	—	—	—	—	—	—
	2	S26	—	—	—	—	—	—	—	—
	3	S26	—	—	—	—	—	—	—	—
	4	S26	—	—	—	—	—	—	—	—
	5	S27	—	—	—	—	—	—	—	—
	6	S29	—	—	—	—	—	—	—	—
	7	S30	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	8	S30	—	—	—	—	—	—	—	—
	9	S32	—	—	—	—	—	—	—	—
	10	S32	—	—	—	—	—	—	—	—
	11	S34	LGH	404	CTG	405	YNN	406	TAC	407
			DT		GGC		KEL		AAT
					CAT				AAT
					GAT				AAG
					ACT				GAG
									CTC
	12	S34	SGH	412	TCT	413	FVK	414	TTT	415
			DN		GGA		ESK		GTG
					CAT				AAA
					GAT				GAG
					AAT				TCT
									AAA
	13	S35	—	—	—	—	—	—	—	—
	14	S35	—	—	—	—	—	—	—	—
	15	S35	SGH	412	TCT	413	FVK	414	TTT	415
			DN		GGA		ESK		GTG
					CAT				AAA
					GAT				GAG
					AAT				TCT
									AAA
	16	S35	—	—	—	—	—	—	—	—
	17	S35	SGH	412	TCT	413	FVK	414	TTT	415
			DN		GGA		ESK		GTG
					CAT				AAA
					GAT				GAG
					AAT				TCT
									AAA
indicates data missing or illegible when filed

TABLE 22
			TCRA
				SEQ		SEQ		SEQ		SEQ
			CDR	ID	CDR	ID	CDR	ID	CDR	ID
	Row	Well	1 (aa)	NO	1 (nt)	NO	2 (aa)	NO	2 (nt)	NO
Donor	1	B1	—	—	—	—	—	—	—	—
2001476	2	C1	—	—	—	—	—	—	—	—
	3	D1	—	—	—	—	—	—	—	—
	4	E1	—	—	—	—	—	—	—	—
	5	F1	—	—	—	—	—	—	—	—
	6	H1	—	—	—	—	—	—	—	—
	7	C2	—	—	—	—	—	—	—	—
	8	D2	—	—	—	—	—	—	—	—
	9	E2	—	—	—	—	—	—	—	—
	10	E3	TISG	424	ACA	425	GLTS	426	GGT	427
			TDY		ATC		N		CTT
					AGT				ACA
					GGA				AGC
					ACT				AAT
					GAT
					TAC
	11	D6	DSV	430	GAC	431	IPSG	432	ATT	433
			NN		TCT		T		CCC
					GTG				TCA
					AAC				GGG
					AAT				ACA
	12	H6	DSV	430	GAC	431	IPSG	432	ATT	433
			NN		TCT		T		CCC
					GTG				TCA
					AAC				GGG
					AAT				ACA
	13	E7	SSVP	333	TCG	434	YTS	335	TAC	336
			PY		TCT		AAT		ACA
					GTT		LV		TCA
					CCA				GCG
					CCG				GCC
					TAT				ACC
									CTG
									GTT
	14	F7	TSGF	435	ACA	436	NVL	437	AAT	438
			NG		TCT		DGL		GTT
					GGG				CTG
					TTC				GAT
					AAC				GGT
					GGG				TTG
	15	B8	DRG	439	GAC	440	IYSN	441	ATA	442
		SQS		CGA		GD		TAC
				GGT				TCC
				TCC				AAT
				CAG				GGT
				TCC				GAC
	16	D8	—	—	—	—	—	—	—	—
	17	F8	—	—	—	—	—	—	—	—
	18	E9	—	—	—	—	—	—	—	—
	19	F9	NSA	408	AAC	409	TYSS	410	ACA	411
			FQY		AGT		GN		TAC
					GCT				TCC
					TTTC				AGT
					AAT				GGT
					AC				AAC
	20	B10	—	—	—	—	—	—	—	—
	21	C10	TISG	424	ACA	425	GLTS	426	GGT	427
			TDY		ATC		N		CTT
					AGT				ACA
					GGA				AGC
					ACT				AAT
					GAT
					TAC
			TCRB
				SEQ		SEQ		SEQ		SEQ
			CDR	ID	CDR	ID	CDR	ID	CDR	ID
	Row	Well	1 (aa)	NO	1 (nt)	NO	2 (aa)	NO	2 (nt)	NO
Donor	1	B1	—	—	—	—	—	—	—	—
2001476	2	C1	—	—	—	—	—	—	—	—
	3	D1	—	—	—	—	—	—	—	—
	4	E1	LGH	416	CTG	417	YSLE	418	TAC	419
			NA		GGT		ER		AGT
					CAT				CTT
					AAC				GAA
					GCT				GAA
									CGG
	5	F1	DFQ	297	GAC	298	SNE	299	TCC	300
			ATT		TTTC		GSK		AAT
					AGG		A		GAG
					CCA				GGC
					CAA				TCC
					CT				AAG
									GCC
	6	H1	—	—	—	—	—	—	—	—
	7	C2	SGH	372	TCT	373	YFSE	374	TAC	375
			RS		GGG		TQ		TTC
					CAT				AGT
					AGG				GAG
					AGT				ACA
									CAG
	8	D2	MGH	420	ATG	421	YSY	422	TAC	423
			RA		GGG		EKL		AGC
					CAC				TAT
					AGG				GAG
					GCT				AAA
									CTC
	9	E2	—	—	—	—	—	—	—	—
	10	E3	SGH	329	TCG	330	FNY	428	TTC	429
			VS		GGT		EAQ		AAT
					CAT				TAT
					GTA				GAA
					TCC				GCC
									CAA
	11	D6	SNH	325	TCT	326	FYN	327	TTTT	328
			LY		AAT		NEI		ATA
					CAC				ATA
					TTA				ATG
					TAC				AAA
									TC
	12	H6	SGH	313	TCT	314	YYE	315	TAT	316
			DT		GGG		EEE		TAT
					CAT				GAG
					GAC				GAG
					ACT				GAA
									GAG
	13	E7	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	14	F7	MNH	355	ATG	356	SVG	357	TCA	358
			EY		AAC		EGT		GTT
					CAT				GGT
					GAA				GAG
					TAC				GGT
									ACA
	15	B8	—	—	—	—	—	—	—	—
	16	D8	—	—	—	—	—	—	—	—
	17	F8	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	18	E9	SGH	293	TCA	294	FNN	295	TTT	296
			NS		GGC		NVP		AAC
					CAC				AAC
					AAC				AAC
					TCC				GTT
									CCG
	19	F9	SGH	362	TCA	363	FNN	295	TTT	296
			DY		GGC		NVP		AAC
					CAC				AAC
					GAC				AAC
					TAC				GTT
									CCG
	20	B10	—	—	—	—	—	—	—	—
	21	C10	SGH	329	TCG	330	FNY	428	TTC	429
			VS		GGT		EAQ		AAT
					CAT				TAT
					GTA				GAA
					TCC				GCC
									CAA

TABLE 23
		TCRA
			SEQ		SEQ		SEQ		SEQ
		CDR	ID	CDR	ID	CDR	ID	CDR	ID
Row	Well	1 (aa)	NO	1 (nt)	NO	2 (aa)	NO	2 (nt)	NO
1	E3	—	—	—	—	—	—	—	—
2	B4	NSM	443	AAC	444	ISSIK	445	ATA	446
		FDY		AGC		DK		AGT
				ATG				TCC
				TTT				ATT
				GAT				AAG
				TAT				GAT
								AAA
3	C4	NSM	443	AAC	444	ISSIK	445	ATA	446
		FDY		AGC		DK		AGT
				ATG				TCC
				TTT				ATT
				GAT				AAG
				TAT				GAT
								AAA
4	E4	—	—	—	—	—	—	—	—
5	F4	—	—	—	—	—	—	—	—
		TCRB
			SEQ		SEQ		SEQ		SEQ
		CDR	ID	CDR	ID	CDR	ID	CDR	ID
Row	Well	1 (aa)	NO	1 (nt)	NO	2 (aa)	NO	2 (nt)	NO
1	E3	—	—	—	—	—	—	—	—
2	B4	SGH	412	TCT	413	FVK	414	TTT	415
		DN		GGA		ESK		GTG
				CAT				AAA
				GAT				GAG
				AAT				TCT
								AAA
3	C4	SGH	412	TCT	413	FVK	414	TTT	415
		DN		GGA		ESK		GTG
				CAT				AAA
				GAT				GAG
				AAT				TCT
								AAA
4	E4	—	—	—	—	—	—	—	—
5	F4	—	—	—	—	—	—	—	—

It will be appreciated by those skilled in the art that changes could be made to the embodiments described above without departing from the broad inventive concept thereof. It is understood, therefore, that this invention is not limited to the particular embodiments disclosed, but it is intended to cover modifications within the spirit and scope of the present invention as defined by the present description.

The following list of embodiments is intended to complement, rather than displace or supersede, the previous descriptions:

Embodiment 1. A capicua transcriptional repressor (CIC) polypeptide fragment comprising an arginine to tryptophan amino acid substitution at a position corresponding to position 215 of SEQ ID NO: 102 (R215W), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02: 01.

Embodiment 2. The CIC polypeptide fragment of embodiment 1, wherein the CIC polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment.

Embodiment 3. The CIC polypeptide fragment of embodiment 1 or 2, wherein the R215W substitution is at amino acid position 8 of the fragment.

Embodiment 4. The CIC polypeptide fragment of any one of embodiments 1-3, wherein the CIC polypeptide fragment is selected from the group consisting of SEQ ID NO: 25, SEQ ID NO: 26 and SEQ ID NO: 27.

Embodiment 5. A catenin beta 1 (CTNNB1) polypeptide fragment comprising: (a) a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C), or (b) a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02:01.

Embodiment 6. The CTNNB1 polypeptide fragment of embodiment 5, wherein the CTNNB1 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment.

Embodiment 7. The CTNNB1 polypeptide fragment of embodiment 5 or 6, wherein the S33C substitution is at amino acid position 4 of the fragment.

Embodiment 8. The CTNNB1 polypeptide fragment of embodiment 5 or 6, wherein the S37F substitution is at amino acid position 8 of the fragment.

Embodiment 9. The CTNNB1 polypeptide fragment of any one of embodiments 5-8, wherein the CTNNB1 polypeptide fragment is selected from the group consisting of SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 80, and SEQ ID NO: 81.

Embodiment 10. An v-erb-b2 erythroblastic leukemia viral oncogene homolog B (ERBB2) polypeptide fragment comprising a valine to isoleucine amino acid substitution at a position corresponding to position 842 of SEQ ID NO: 104 (V842I), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02:01.

Embodiment 11. The ERBB2 polypeptide fragment of embodiment 10, wherein the ERBB2 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment.

Embodiment 12. The ERBB2 polypeptide fragment of embodiment 10 or 11, wherein the V8421 substitution at amino acid position 3 of the fragment.

Embodiment 13. The ERBB2 polypeptide fragment of any one of embodiments 10-13, wherein the ERBB2 polypeptide fragment is selected from the group consisting of SEQ ID NO: 84, SEQ ID NO: 85, and SEQ ID NO: 86.

Embodiment 14. A kirsten rat sarcoma (KRAS) polypeptide fragment comprising: (a) a glycine to alanine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12A), (b) a glycine to cysteine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12C), or (c) a glycine to valine amino acid substitution at a position corresponding to at position 12 of SEQ ID NO: 105 (G12V), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02:01.

Embodiment 15. The KRAS polypeptide fragment of embodiment 14, wherein the KRAS polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment.

Embodiment 16. The KRAS polypeptide fragment of embodiment 14 or 15, wherein the G12A substitution is at amino acid position 7 of the fragment.

Embodiment 17. The KRAS polypeptide fragment of embodiment 14 or 15, wherein the G12C substitution is at amino acid position 7 of the fragment.

Embodiment 18. The KRAS polypeptide fragment of embodiment 14 or 5, wherein the G12V substitution is at amino acid position 7 of the fragment.

Embodiment 19. The KRAS polypeptide fragment of any one of embodiments 14-18, wherein the KRAS polypeptide fragment is selected from the group consisting of SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, and SEQ ID NO: 42.

Embodiment 20. A phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) polypeptide fragment comprising: (a) a glutamic acid to lysine amino acid substitution at a position corresponding to position 453 of SEQ ID NO: 106 (E453K), or (b) a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02:01.

Embodiment 21. The PIK3CA polypeptide fragment of embodiment 20, wherein the PIK3CA polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment.

Embodiment 22. The PIK3CA polypeptide fragment of embodiment 20 or 21, wherein the E453K substitution is at amino acid position 3 of the fragment.

Embodiment 23. The PIK3CA polypeptide fragment of embodiment 20 or 21, wherein the G118D substitution is at amino acid position 7 of the fragment.

Embodiment 24. The PIK3CA polypeptide fragment of any one of embodiments 20-23, wherein the PIK3CA polypeptide fragment is selected from the group consisting of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, and SEQ ID NO: 47.

Embodiment 25. A phosphatase and tensin homolog (PTEN) polypeptide fragment comprising an arginine to cysteine amino acid substitution at a position corresponding to position 173 of SEQ ID NO: 107 (R173C), and an amino acid substitution at amino acid position 3 of the fragment, amino acid position 10 of the fragment, or both, wherein the fragment is ten amino acids in length, and wherein the fragment binds to HLA-A*02:01.

Embodiment 26. The PTEN polypeptide fragment of embodiment 25, wherein the PTEN polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment.

Embodiment 27. The PTEN polypeptide fragment of embodiment 25 or 26, wherein the R173C substitution is at amino acid position 1 of the fragment.

Embodiment 28. The PTEN polypeptide fragment of any one of embodiments 25-27, wherein the PTEN polypeptide fragment is selected from the group consisting of SEQ ID NO: 48, SEQ ID NO: 49, and SEQ ID NO: 88.

Embodiment 29. A splicing factor 3b subunit 1 (SF3B1) polypeptide fragment comprising an arginine to histidine amino acid substitution at a position corresponding to position 625 of SEQ ID NO: 108 (R625H), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02: 01.

Embodiment 30. The SF3B1 polypeptide fragment of embodiment 29, wherein the SF3B1 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native epitope.

Embodiment 31. The SF3B1 polypeptide fragment of embodiment 29 or 30, wherein the R625H substitution is at amino acid position 7 of the fragment.

Embodiment 32. The SF3B1 polypeptide fragment of any one of embodiments 29-31, wherein the SF3B1 polypeptide fragment is selected from the group consisting of SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 90, SEQ ID NO: 91 and SEQ ID NO: 92.

Embodiment 33. A SRY-box transcription factor 17 (SOX17) polypeptide fragment comprising a serine to isoleucine amino acid substitution at a position corresponding to position 403 of SEQ ID NO: 109 (S403I), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02: 01.

Embodiment 34. The SOX17 polypeptide fragment of embodiment 33, wherein the SOX17 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment.

Embodiment 35. The SOX17 polypeptide fragment of embodiment 33 or 34, wherein the S403I substitution is at amino acid position 6 of the fragment.

Embodiment 36. The SOX17 polypeptide fragment of any one of embodiments 33-35, wherein the SOX17 polypeptide fragment is selected from the group consisting of SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, and SEQ ID NO: 93.

Embodiment 37. A tumor protein 53 (TP53) polypeptide fragment comprising: (a) an arginine to leucine amino acid substitution at a position corresponding to position 110 of SEQ ID NO: 110 (R110L), (b) a serine to phenylalanine amino acid substitution at a position corresponding to position 127 of SEQ ID NO: 110 (S127F), (c) a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N), (d) a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y), (e) a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L), (f) a histidine to leucine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193L), (g) a histidine to tyrosine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193Y), (h) a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C), or (i) a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M), and an amino acid substitution at amino acid position 2 of the fragment, amino acid position 9 of the fragment, or both, wherein the fragment is at least nine amino acids in length, and wherein the fragment binds to HLA-A*02:01.

Embodiment 38. The TP53 polypeptide fragment of embodiment 37, wherein the TP53 polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment.

Embodiment 39. The TP53 polypeptide fragment of embodiment 37 or 38, wherein the R110L substitution is at amino acid position 8 of the fragment.

Embodiment 40. The TP53 polypeptide fragment of embodiment 37 or 38, wherein the S127F substitution is at amino acid position 7 of the fragment.

Embodiment 41. The TP53 polypeptide fragment of embodiment 37 or 38, wherein the K132N substitution is at amino acid position 4 of the fragment.

Embodiment 42. The TP53 polypeptide fragment of embodiment 37 or 38, wherein the C141Y substitution is at amino acid position 3 of the fragment.

Embodiment 43. The TP53 polypeptide fragment of embodiment 37 or 38, wherein the P152L substitution is at amino acid position 9 of the fragment.

Embodiment 44. The TP53 polypeptide fragment of embodiment 37 or 38, wherein the H193L substitution is at amino acid position 7 of the fragment.

Embodiment 45. The TP53 polypeptide fragment of embodiment 37 or 38, wherein the Y220C substitution is at amino acid position 4 of the fragment.

Embodiment 46. The TP53 polypeptide fragment of embodiment 37 or 38, wherein the V272M substitution is at amino acid position 9 of the fragment.

Embodiment 47. The TP53 polypeptide fragment of any one of embodiments 37-46, wherein the TP53 polypeptide fragment is selected from the group consisting of SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 94, SEQ ID NO: 95, and SEQ ID NO: 96.

Embodiment 48. A polypeptide fragment selected from the group consisting of SEQ ID NO: 25, SEQ ID NO: 26 and SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, and SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81; SEQ ID NO: 84, SEQ ID NO: 85, and SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 91,SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, and SEQ ID NO: 96.

Embodiment 49. A polypeptide fragment selected from the group consisting of SEQ ID NO: 29, SEQ ID NO: 32, SEQ ID NO: 45, SEQ ID NO: 59, SEQ ID NO: 64, SEQ ID NO: 68, SEQ ID NO: 75 and SEQ ID NO: 78.

Embodiment 50. A polynucleotide encoding at least one or more polypeptide fragments according to any one of embodiment 1-49.

Embodiment 51. The polynucleotide of embodiment 50, wherein the polynucleotide is cDNA.

Embodiment 52. A vector comprising at least one or more polynucleotides of embodiment 50 or 51.

Embodiment 53. The vector of embodiment 52, wherein the vector is selected from an adenovirus vector, an alphaviral vector, a poxvirus vector, an adeno-associated virus vector, a retrovirus vector, a self-replicating RNA molecule, and a combination thereof.

Embodiment 54. The vector of embodiment 53, wherein the adenovirus vector is selected from hAd5, hAd7, hAd11, hAd26, hAd34, hAd35, hAd48, hAd49, hAd50, GAd20, Gad19, GAd21, GAd25, GAd26, GAd27, GAd28, GAd29, GAd30, GAd31, ChAd3, ChAd4, ChAd5, ChAd6, ChAd7, ChAd8, ChAd9, ChAd10, ChAd11, ChAd16, ChAdI7, ChAd19, ChAd20, ChAd22, ChAd24, ChAd26, ChAd30, ChAd31, ChAd37, ChAd38, ChAd44, ChAd55, ChAd63, ChAd73, ChAd82, ChAd83, ChAd146, ChAd147, PanAd1, PanAd2, and PanAd3.

Embodiment 55. The vector of embodiment 53, wherein the poxvirus vector is selected from smallpox virus vector, vaccinia virus vector, cowpox virus vector, monkeypox virus vector, Copenhagen vaccinia virus (W) vector, New York Attenuated Vaccinia Virus (NYVAC) vector, and Modified Vaccinia Ankara (MVA) vector.

Embodiment 56. The vector of embodiment 53, wherein the vector is the adenovirus vector comprising a polynucleotide encoding at least one or more polypeptide fragments according to any one of embodiment 1-55.

Embodiment 57. A pharmaceutical composition comprising at least one or more polypeptide fragments according to any one of embodiments 1-49.

Embodiment 58. A pharmaceutical composition comprising a polynucleotide according to embodiment 50 or 51.

Embodiment 59. A pharmaceutical composition comprising a vector according to any one of embodiments 52-56.

Embodiment 60. A method of treating cancer in a subject comprising administering to the subject in need thereof the polypeptide of any one of embodiments 1-49, the polynucleotide of embodiment 50 or 51, the vector of any one of embodiments 52-56, or the pharmaceutical composition of any one of embodiments 57-59.

Embodiment 61. A method of inducing an immune response in a subject comprising administering to the subject in need thereof the polypeptide of any one of embodiments 1-49, the polynucleotide of embodiment 50 or 51, the vector of any one of embodiments 52-56, or the pharmaceutical composition of any one of embodiments 57-59.

Embodiment 62. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a catenin beta 1 (CTNNB1) mutant comprising a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C) in a subject, the method comprising administering to the subject in need thereof (a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 2, (b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 29, or (c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Embodiment 63. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a catenin beta 1 (CTNNB1) mutant comprising a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F) in a subject, the method comprising administering to the subject in need thereof (a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 3, (b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 32, or (c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Embodiment 64. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) mutant comprising a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D) in a subject, the method comprising administering to the subject in need thereof (a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 9, (b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 45, or (c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Embodiment 65. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y) in a subject, the method comprising administering to the subject in need thereof (a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 13, (b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 59, or (c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Embodiment 66. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N) in a subject, the method comprising administering to the subject in need thereof (a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 16, (b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 64, or (c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Embodiment 67. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L) in a subject, the method comprising administering to the subject in need thereof (a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 18, (b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 68, or (c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Embodiment 68. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M) in a subject, the method comprising administering to the subject in need thereof (a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 22, (b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 75, or (c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Embodiment 69. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C) in a subject, the method comprising administering to the subject in need thereof (a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 23, (b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 78, or (c) a combination thereof in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

Embodiment 70. The method of any one of embodiments 62-69, comprising administering the polynucleotide in part a) prior to administering the polynucleotide in part b).

Embodiment 71. The method of any one of embodiments 62-69, comprising administering the polynucleotide in part b) prior to administering the polynucleotide in part a).

Embodiment 72. The method of any one of embodiments 62-71, comprising administering the polynucleotide in part a) concurrently with the polynucleotide in part b).

Embodiment 73. The method of any one of embodiments 62-69, comprising administering a vector encoding the polynucleotide of part a) and a vector encoding the polynucleotide of part b).

Embodiment 74. The method of embodiment 73, wherein the vectors are independently selected from an adenovirus vector, an alphaviral vector, a poxvirus vector, an adeno-associated virus vector, a retrovirus vector, and a self-replicating RNA molecule.

Embodiment 75. The method of embodiment 74, wherein the adenovirus vector is selected from hAd5, hAd7, hAd11, hAd26, hAd34, hAd35, hAd48, hAd49, hAd50, GAd20, Gad19, GAd21, GAd25, GAd26, GAd27, GAd28, GAd29, GAd30, GAd31, ChAd3, ChAd4, ChAd5, ChAd6, ChAd7, ChAd8, ChAd9, ChAd10, ChAd11, ChAd16, ChAdI7, ChAd19, ChAd20, ChAd22, ChAd24, ChAd26, ChAd30, ChAd31, ChAd37, ChAd38, ChAd44, ChAd55, ChAd63, ChAd73, ChAd82, ChAd83, ChAd146, ChAd147, PanAd1, PanAd2, and PanAd3.

Embodiment 76. The method of embodiment 74, wherein the poxvirus vector is selected from smallpox virus vector, vaccinia virus vector, cowpox virus vector, monkeypox virus vector, Copenhagen vaccinia virus (W) vector, New York Attenuated Vaccinia Virus (NYVAC) vector, and Modified Vaccinia Ankara (MVA) vector.

Embodiment 77. A kit of parts comprising a pair of polypeptide fragments selected from the group consisting of: (a) SEQ ID NO: 2 and SEQ ID NO: 29; (b) SEQ ID NO: 3 and SEQ ID NO: 32; (c) SEQ ID NO: 9 and SEQ ID NO: 45; (d) SEQ ID NO: 13 and SEQ ID NO: 59; (e) SEQ ID NO: 16 and SEQ ID NO: 64; (f) SEQ ID NO: 18 and SEQ ID NO 68; (g) SEQ ID NO: 22 and SEQ ID NO: 75; and (h) SEQ ID NO: 23 and SEQ ID NO: 78.

Embodiment 78. A kit of parts comprising a pair of polypeptide fragments selected from the group consisting of: (a) SEQ ID NO: 9 and SEQ ID NO: 45 (b) SEQ ID NO: 13 and SEQ ID NO: 59; and (c) SEQ ID NO: 18 and SEQ ID NO 68.

Embodiment 79. A method for generating CD8+ T-cells that are positive for an HLA-A*02:01-restricted polypeptide fragment and a cognate native polypeptide fragment, comprising exposing CD8+ T-cells to the HLA-A*02:01-restricted polypeptide fragment and cognate native polypeptide fragment selected from the group consisting of: (a) SEQ ID NO: 2 and SEQ ID NO: 29; (b) SEQ ID NO: 3 and SEQ ID NO: 32; (c) SEQ ID NO: 9 and SEQ ID NO: 45; (d) SEQ ID NO: 13 and SEQ ID NO: 59; (e) SEQ ID NO: 16 and SEQ ID NO: 64; (f) SEQ ID NO: 18 and SEQ ID NO 68; (g) SEQ ID NO: 22 and SEQ ID NO: 75; and (h) SEQ ID NO: 23 and SEQ ID NO: 78; and selecting CD8+ T cells that are positive to both the HLA-A*02:01-restricted polypeptide fragment and a cognate neoantigen polypeptide fragment.

Embodiment 80. A method for generating CD8+ T-cells that are positive for an HLA-A*02:01-restricted polypeptide fragment and a cognate native polypeptide fragment, comprising exposing CD8+ T-cells to the HLA-A*02:01-restricted polypeptide fragment and cognate native polypeptide fragment selected from the group consisting of: (a) SEQ ID NO: 9 and SEQ ID NO: 45; (b) SEQ ID NO: 13 and SEQ ID NO: 59; and (c) SEQ ID NO: 18 and SEQ ID NO 68; and selecting CD8+ T cells that are positive to both the HLA-A*02:01-restricted polypeptide fragment and a cognate neoantigen polypeptide fragment.

Embodiment 81. A T-cell receptor (TCR) comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 120 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (b) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 118 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (c) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 122 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (d) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 122 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 124; (e) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 122 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 134; (f) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 112 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (g) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 116 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (h) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 116 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 128; (i) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 126 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (j) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (k) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 132; or (l) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 134.

Embodiment 82. A T-cell receptor (TCR) comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 118 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (b) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 118 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 142; (c) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 207 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (d) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 112 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (e) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 205 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (f) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (g) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 166; (h) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 186 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (i) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 152 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (j) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (k) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 142; (l) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 150; (m) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 162; (n) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 136 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (o) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 136 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 138; (p) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 136 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 142; (q) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 140 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (r) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 140 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 160; (s) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 144 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (t) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 144 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 146; (u) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 144 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 158; (v) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 148 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (w) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 148 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 150; (x) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 154 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 156; (y) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (z) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 166; (aa) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 180; (bb) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 182; (cc) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 197; (dd) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 168 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (ee) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 168 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 170; (ff) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 168 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 199; (gg) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 174 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 176; (hh) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 174 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 178; (ii) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 184 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (jj) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 184 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 188; (kk) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 190 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 192; (ll) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 194 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (mm) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 201 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 203; or (nn) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 210 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114.

Embodiment 83. A T-cell receptor (TCR) comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (b) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 170; (c) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 213; (d) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 152 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (e) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 168 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 170; (f) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 215 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (g) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 217 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (h) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 219 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (i) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 219 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 221; or (j) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 223 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 221.

Embodiment 84. A T-cell receptor (TCR) comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 152 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 252; (b) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 152 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (c) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 248 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 250; (d) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 248 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 258; (e) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 254 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 256; (f) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 254 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (g) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 263 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 265; (h) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 267 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 269; (i) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 267 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 271; (j) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 273 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 275; (k) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 277 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 279; (l) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 281 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 283; or (m) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 285 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 287.

Embodiment 85. A T-cell receptor (TCR) comprising an alpha chain and a beta chain, wherein: (a) the alpha chain comprises a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 248 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114; (b) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 289 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 291; or (c) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 289 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114.

Embodiment 86. A polynucleotide encoding the TCR of any one of embodiments 81-85.

Embodiment 87. A vector comprising the polynucleotide of embodiment 86.

Embodiment 88. A cell transformed to express the polynucleotide of embodiment 86.

Embodiment 89. A cell comprising the vector of embodiment 87.

Embodiment 90. The cell of embodiment 88 or 89, wherein the cell is a CD8+ T cell.

Embodiment 91. A pharmaceutical composition comprising the TCR of any one of embodiment 82-85, the polynucleotide of embodiment 86, the vector of embodiment 87, or the cell of any one of embodiments 88-90.

Embodiment 92. A method of treating cancer in a subject comprising administering to the subject in need thereof a pharmaceutical composition of embodiment 91.

Embodiment 93. A method of inducing an immune response in a subject comprising administering to the subject in need thereof a pharmaceutical composition of embodiment 91.

Embodiment 94. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) mutant comprising a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D) in a subject comprising administering to the subject in need thereof the TCR of embodiment 81.

Embodiment 95. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y) in a subject comprising administering to the subject in need thereof the TCR of embodiment 82.

Embodiment 96. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L) in a subject comprising administering to the subject in need thereof the TCR of embodiment 83.

Embodiment 97. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a catenin beta 1 (CTNNB1) mutant comprising a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F) in a subject comprising administering to the subject in need thereof the TCR of embodiment 84.

Embodiment 98. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition characterized by expression of a tumor protein 53 (TP53) mutant comprising a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C) in a subject comprising administering to the subject in need thereof the TCR of embodiment 85.

Claims

1. A polypeptide fragment selected from: a) a capicua transcriptional repressor (CIC) polypeptide fragment comprising an arginine to tryptophan amino acid substitution at a position corresponding to position 215 of SEQ ID NO: 102 (R215W), wherein the R215W substitution is at amino acid position 8 of the fragment;b) a catenin beta 1 (CTNNB1) polypeptide fragment comprising: i. a serine to cysteine amino acid substitution at a position corresponding to position 33 of SEQ ID NO: 103 (S33C), wherein the S33C substitution is at amino acid position 4 of the fragment, orii. a serine to phenylalanine amino acid substitution at a position corresponding to position 37 of SEQ ID NO: 103 (S37F), wherein the S37F substitution is at amino acid position 8 of the fragment,c) a v-erb-b2 erythroblastic leukemia viral oncogene homolog B (ERBB2) polypeptide fragment comprising a valine to isoleucine amino acid substitution at a position corresponding to position 842 of SEQ ID NO: 104 (V842I), wherein the V842I substitution at amino acid position 3 of the fragmentd) kirsten rat sarcoma (KRAS) polypeptide fragment comprising: i. a glycine to alanine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12A), wherein the G12A substitution is at amino acid position 7 of the fragment,ii. a glycine to cysteine amino acid substitution at a position corresponding to position 12 of SEQ ID NO: 105 (G12C), wherein the G12C substitution is at amino acid position 7 of the fragment, oriii. a glycine to valine amino acid substitution at a position corresponding to at position 12 of SEQ ID NO: 105 (G12V), wherein the G12V substitution is at amino acid position 7 of the fragment,e) a phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) polypeptide fragment comprising: i. a glutamic acid to lysine amino acid substitution at a position corresponding to position 453 of SEQ ID NO: 106 (E453K), wherein the E453K substitution is at amino acid position 3 of the fragment, orii. a glycine to aspartic acid amino acid substitution at a position corresponding to position 118 of SEQ ID NO: 106 (G118D), wherein the G118D substitution is at amino acid position 7 of the fragment,f) a phosphatase and tensin homolog (PTEN) polypeptide fragment comprising an arginine to cysteine amino acid substitution at a position corresponding to position 173 of SEQ ID NO: 107 (R173C), wherein the R173C substitution is at amino acid position 1 of the fragmentg) a splicing factor 3b subunit 1 (SF3B1) polypeptide fragment comprising an arginine to histidine amino acid substitution at a position corresponding to position 625 of SEQ ID NO: 108 (R625H), wherein the R625H substitution is at amino acid position 7 of the fragmenth) a SRY-box transcription factor 17 (SOX17) polypeptide fragment comprising a serine to isoleucine amino acid substitution at a position corresponding to position 403 of SEQ ID NO: 109 (S403I), wherein the S403I substitution is at amino acid position 6 of the fragmenti) a tumor protein 53 (TP53) polypeptide fragment comprising: a) an arginine to leucine amino acid substitution at a position corresponding to position 110 of SEQ ID NO: 110 (R110L), wherein the R110L substitution is at amino acid position 8 of the fragment,b) a serine to phenylalanine amino acid substitution at a position corresponding to position 127 of SEQ ID NO: 110 (S127F), wherein the S127F substitution is at amino acid position 7 of the fragment,c) a lysine to asparagine amino acid substitution at a position corresponding to position 132 of SEQ ID NO: 110 (K132N), wherein the K132N substitution is at amino acid position 4 of the fragment,d) a cysteine to tyrosine amino acid substitution at a position corresponding to position 141 of SEQ ID NO: 110 (C141Y), wherein the C141Y substitution is at amino acid position 3 of the fragment,e) a proline to leucine amino acid substitution at a position corresponding to position 152 of SEQ ID NO: 110 (P152L), wherein the P152L substitution is at amino acid position 9 of the fragment,f) a histidine to leucine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193L), wherein the H193L substitution is at amino acid position 7 of the fragment,g) a histidine to tyrosine amino acid substitution at a position corresponding to position 193 of SEQ ID NO: 110 (H193Y), wherein the H193L substitution is at amino acid position 7 of the fragment,h) a tyrosine to cysteine amino acid substitution at a position corresponding to position 220 of SEQ ID NO: 110 (Y220C), wherein the Y220C substitution is at amino acid position 4 of the fragment, ori) a valine to methionine amino acid substitution at a position corresponding to position 272 of SEQ ID NO: 110 (V272M), wherein the V272M substitution is at amino acid position 9 of the fragment

2. The polypeptide fragment of claim 1, wherein the polypeptide fragment has greater affinity for HLA-A*02:01 than a cognate native polypeptide fragment.

3. A polypeptide fragment selected from the group consisting of SEQ ID NO: 25, SEQ ID NO: 26 and SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, and SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81; SEQ ID NO: 84, SEQ ID NO: 85, and SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 91,SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, and SEQ ID NO: 96.

4. A polypeptide fragment selected from the group consisting of SEQ ID NO: 29, SEQ ID NO: 32, SEQ ID NO: 45, SEQ ID NO: 59, SEQ ID NO: 64, SEQ ID NO: 68, SEQ ID NO: 75 and SEQ ID NO: 78.

5. A polynucleotide encoding at least one or more polypeptide fragments according to claim 1.

6. The polynucleotide of claim 5, wherein the polynucleotide is cDNA.

7. A vector comprising at least one or more polynucleotides of claim 5.

8. The vector of claim 7, wherein the vector is selected from an adenovirus vector, an alphaviral vector, a poxvirus vector, an adeno-associated virus vector, a retrovirus vector, a self-replicating RNA molecule, and a combination thereof

9. A pharmaceutical composition comprising at least one or more polypeptide fragments according to claim 1.

10. A method of treating cancer in a subject comprising administering to the subject in need thereof the polypeptide of claim 1.

11. A method of inducing an immune response in a subject comprising administering to the subject in need thereof the polypeptide of claim 1.

12. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition, the method comprising administering to the subject in need thereof a) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 2 and/or a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 29,b) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 3 and/or a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 32,c) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 9 and/or a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 45,d) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 13 and/or a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 59,e) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 16 and/or a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 64,f) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 18 and/or a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 68,g) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 22 and/or a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 75, orh) a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 23 and/or a polynucleotide encoding a polypeptide fragment comprising SEQ ID NO: 78, in an amount effective to treat, prevent, reduce the risk of onset or delay the onset of the clinical condition.

13. The method of claim 12, comprising administering a vector encoding the polynucleotide.

14. The method of claim 13, wherein the vectors are independently selected from an adenovirus vector, an alphaviral vector, a poxvirus vector, an adeno-associated virus vector, a retrovirus vector, and a self-replicating RNA molecule.

15. A kit of parts comprising a pair of polypeptide fragments selected from the group consisting of: a. SEQ ID NO: 2 and SEQ ID NO: 29;b. SEQ ID NO: 3 and SEQ ID NO: 32;c. SEQ ID NO: 9 and SEQ ID NO: 45;d. SEQ ID NO: 13 and SEQ ID NO: 59;e. SEQ ID NO: 16 and SEQ ID NO: 64;f SEQ ID NO: 18 and SEQ ID NO 68;g. SEQ ID NO: 22 and SEQ ID NO: 75; andh. SEQ ID NO: 23 and SEQ ID NO: 78.

16. A method for generating CD8+ T-cells that are positive for an HLA-A*02:01-restricted polypeptide fragment and a cognate native polypeptide fragment, comprising exposing CD8+ T-cells to the HLA-A*02:01-restricted polypeptide fragment and cognate native polypeptide fragment selected from the group consisting of: a. SEQ ID NO: 2 and SEQ ID NO: 29;b. SEQ ID NO: 3 and SEQ ID NO: 32;c. SEQ ID NO: 9 and SEQ ID NO: 45;d. SEQ ID NO: 13 and SEQ ID NO: 59;e. SEQ ID NO: 16 and SEQ ID NO: 64;f. SEQ ID NO: 18 and SEQ ID NO 68;g. SEQ ID NO: 22 and SEQ ID NO: 75; andh. SEQ ID NO: 23 and SEQ ID NO: 78;

17. A T-cell receptor (TCR) comprising an alpha chain and a beta chain, wherein: a. the alpha chain comprises a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 120 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;b. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 118 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;c. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 122 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;d. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 122 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 124;e. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 122 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 134;f. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 112 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;g. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 116 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;h. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 116 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 128;i. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 126 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;j. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;k. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 132; orl. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 134.m. the alpha chain comprises a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 118 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;n. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 118 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 142;o. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 207 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;p. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 112 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;q. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 205 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;r. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;s. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 166;t. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 186 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;u. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 152 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;v. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;w. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 142;x. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 150;y. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 130 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 162;z. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 136 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;aa. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 136 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 138;bb. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 136 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 142;cc. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 140 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;dd. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 140 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 160;ee. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 144 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;ff. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 144 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 146;gg. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 144 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 158;hh. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 148 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;ii. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 148 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 150;jj. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 154 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 156;kk. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;ll. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 166;mm. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 180;nn. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 182;oo. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 164 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 197;pp. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 168 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;qq. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 168 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 170;rr. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 168 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 199;ss. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 174 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 176;tt. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 174 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 178;uu. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 184 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;vv. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 184 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 188;ww. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 190 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 192;xx. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 194 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;yy. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 201 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 203; orzz. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 210 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114.aaa. the alpha chain comprises a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;bbb. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 170;ccc. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 172 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 213;ddd. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 152 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;eee. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 168 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 170;fff. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 215 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;ggg. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 217 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;hhh. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 219 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;iii. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 219 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 221; orjjj. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 223 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 221.kkk. the alpha chain comprises a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 152 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 252;lll. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 152 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;mmm. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 248 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 250;nnn. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 248 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 258;ooo. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 254 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 256;ppp. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 254 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;qqq. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 263 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 265;rrr. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 267 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 269;sss. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 267 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 271;ttt. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 273 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 275;uuu. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 277 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 279;vvv. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 281 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 283; orwww. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 285 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 287.xxx. the alpha chain comprises a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 248 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114;yyy. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 289 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 291; orzzz. the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 289 and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 114.

18. A polynucleotide encoding the TCR of claim 17.

19. A vector comprising the polynucleotide of claim 18.

20. A cell transformed to express the polynucleotide of claim 19.

21. A cell comprising the vector of claim 17.

22. A method of treating, preventing, reducing a risk of onset or delaying the onset of a clinical condition in a subject comprising administering to the subject in need thereof the TCR of claim 17.