Patent Application
20230079750
The instant application contains a sequence listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Feb. 24, 2022, is named JBI6418USNP1_SL.txt and is 511,902 bytes in size.
Provided are T-cell receptors (TCRs) that bind to CALR or JAK2 antigens. Also provided are TCRs, polynucleotides and vectors that encode the TCRs, cells comprising the TCRs, and methods of treatment.
Janus Kinase 2 (JAK2) is a cytoplasmic protein involved in a signaling pathway known to regulate cell growth and proliferation, mainly through cytokine receptors on the cell surface. This pathway is important for generation and differentiation of blood cells from hemopoietic stem cells in the bone marrow. Lack of JAK2 is embryonic lethal due to lack of erythropoiesis (red blood cell production). JAK2 mutations have been correlated with myeloproliferative neoplasms (MPNs), such as polycythemia vera (PV), essential thromocythemia (ET), and primary myelofibrosis (PM). The most clinically relevant mutation is V617F, where the valine at amino acid position 617 is replaced by the amino acid phenylalanine. This mutation is an activating mutation making cells more sensitive to growth factors, leading to increased blood cell proliferation in these diseases. The prevalence of this mutation in MPN is high. Almost 100% of PV, 50% of PM, and 60% of ET patients harbor this mutation. The current standard of care treatment for MPN is Ruxolitinib, a JAK inhibitor, but this treatment is not curative. An unmet medical need still remains for treatment of these diseases.
Described herein are T-cell receptors (TCRs) comprising an alpha chain and a beta chain, wherein the alpha chain comprises a complementarity determining region 3 (CDR3) comprising an amino acid sequence provided in Table 4, and the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 5 (i.e. the beta chain CDR3 provided in Table 5 is in the same row number (row 1, for example) as the alpha chain CDR3 provided in Table 4 (row 1, for example)).
Also described herein are TCRs comprising an alpha chain and a beta chain, wherein the alpha chain comprises a complementarity determining region 1 (CDR1) comprising an amino acid sequence provided in Table 4, a complementarity determining region 2 (CDR2) comprising an amino acid sequence provided in Table 4, and a CDR3 comprising an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 5, a CDR2 comprising an amino acid sequence provided in Table 5, and a CDR3 comprising an amino acid sequence provided in Table 5 (i.e. the beta chain CDRs provided in Table 5 are in the same row number (row 1, for example) as the alpha chain CDRs provided in Table 4 (row 1, for example)).
In certain embodiments, the alpha chain comprises a variable and joining (VJ) region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a variable, diversity and joining (VDJ) region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5. An alpha chain VJ region corresponds to a beta chain VDJ region if they appear in the same row in Table 4 and Table 5 (i.e. the beta chain VDJ region provided in Table 5 is in the same row number (row 1, for example) as the alpha chain VJ region provided in Table 4 (row 1, for example)).
In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5 (i.e. the beta chain provided in Table 5 is in the same row number (row 1, for example) as the alpha chain provided in Table 4 (row 1, for example)).
Also described herein are nucleic acid molecules encoding the disclosed TCRs.
Also described herein are vectors comprising a disclosed nucleic acid molecule.
Further described provided are cells transformed to express any of the nucleic acid molecules described herein.
Still further described are cells comprising any of the disclosed vectors. In certain embodiments, the cell is a CD8+ T cell.
Also provided are pharmaceutical compositions comprising any of the disclosed TCRs, any of the disclosed nucleic acid molecules, any of the disclosed vectors, or any of the disclosed cells.
The disclosed T-cell receptors (TCRs), polynucleotides, vectors, compositions, kits, and cells may be understood more readily by reference to the following detailed description, which forms a part of this disclosure. It is to be understood that the disclosed TCRs, polynucleotides, vectors, compositions, kits, 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 TCRs, polynucleotides, vectors, compositions, kits, 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 TCRs, polynucleotides, vectors, compositions, kits, 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.
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.
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.
“9-mer” or “9mer” refers to a polypeptide that is nine amino acids in length.
“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.
Disclosed herein are TCRs that bind to CALR and JAK2 epitopes. In some embodiments, the TCRs bind to a JAK2 epitope comprising the amino acid sequence of VLNYGVCFC (SEQ ID NO: 681). In some embodiments, the TCRs bind to a JAK2 epitope comprising the amino acid sequence of FCGDENILV (SEQ ID NO: 682). In some embodiments, the TCRs bind to a CALR epitope contained within the amino acid sequence
MKDKQDEEQRTRRMMRTKMRMRRMRRTRRKMRRKMSPARPRTSCREACLQ
GWTEAAYEEAEDNCRRMMRTKAAYVLNYGVCFCAAYFCGDENILV,
wherein the mutant CALR type 1 sequence is underlined, the mutant CALR type II sequence is underlined and bold, and the mutant JAK2 sequences are underlined an italics. In some embodiments, the TCRs bind to a mutJAK2 and mutCALR antigen construct comprising an amino acid sequence of SEQ ID NO: 702.
The disclosed TCRs may be identified based on T-cell binding to the CALR and JAK2 epitopes, followed by sequencing of the TCR. The TCRs may be obtained from αβ T cells. The 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 a 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, or domain swapped with α and β chain variable and/or constant domains swapped as described in U.S. Pat. Nos. 7,329,731, 7,569,664, 9,133,264, 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 the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 4, and the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 5. (i.e. the beta chain CDR3 provided in Table 5 is in the same row number (row 1, for example) as the alpha chain CDR3 provided in Table 4 (row 1, for example)).
Also described herein are TCRs comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 4, a CDR2 comprising an amino acid sequence provided in Table 4, and a CDR3 comprising an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 5, a CDR2 comprising an amino acid sequence provided in Table 5, and a CDR3 comprising an amino acid sequence provided in Table 5 (i.e. the beta chain CDRs provided in Table 5 are in the same row number (row 1, for example) as the alpha chain CDRs provided in Table 4 (row 1, for example)).
In certain embodiments, the alpha chain comprises a variable and joining (VJ) region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a variable, diversity and joining (VDJ) region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5 (i.e. the beta chain VDJ region provided in Table 5 is in the same row number (row 1, for example) as the alpha chain VJ region provided in Table 4 (row 1, for example)).
In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5 (i.e. the beta chain provided in Table 5 is in the same row number (row 1, for example) as the alpha chain provided in Table 4 (row 1, for example)).
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 7, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 245. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 3, a CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 7, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 241, a CDR2 comprising the amino acid sequence of SEQ ID NO: 243, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 245. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 479, and the beta chain comprises a VDJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 581. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 477, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 579.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 14, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 252. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 10, a CDR2 comprising the amino acid sequence of SEQ ID NO: 12, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 14, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 248, a CDR2 comprising the amino acid sequence of SEQ ID NO: 250, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 252. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 482, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 584. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 480, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 582.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 21, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 259. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 17, a CDR2 comprising the amino acid sequence of SEQ ID NO: 19, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 21, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 255, a CDR2 comprising the amino acid sequence of SEQ ID NO: 257, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 259. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 485, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 587. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 483, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 585.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 28, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 266. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 24, a CDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 28, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 262, a CDR2 comprising the amino acid sequence of SEQ ID NO: 264, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 266. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 488, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 590. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 486, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 588.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 35, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 273. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 31, a CDR2 comprising the amino acid sequence of SEQ ID NO: 33, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 35, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 269, a CDR2 comprising the amino acid sequence of SEQ ID NO: 271, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 273. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 491, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 593. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 489, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 591.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 42, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 280. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 38, a CDR2 comprising the amino acid sequence of SEQ ID NO: 40, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 42, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 276, a CDR2 comprising the amino acid sequence of SEQ ID NO: 278, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 280. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 494, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 596. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 492, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 594.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 49, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 287. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a CDR2 comprising the amino acid sequence of SEQ ID NO: 47, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 49, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 283, a CDR2 comprising the amino acid sequence of SEQ ID NO: 285, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 287. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 497, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 599. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 495, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 597.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 56, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 294. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 52, a CDR2 comprising the amino acid sequence of SEQ ID NO: 54, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 56, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 290, a CDR2 comprising the amino acid sequence of SEQ ID NO: 292, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 294. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 500, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 602. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 498, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 600.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 63, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 301. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 59, a CDR2 comprising the amino acid sequence of SEQ ID NO: 61, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 63, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 297, a CDR2 comprising the amino acid sequence of SEQ ID NO: 299, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 301. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 503, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 605. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 501, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 603.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 70, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 308. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 66, a CDR2 comprising the amino acid sequence of SEQ ID NO: 68, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 70, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 304, a CDR2 comprising the amino acid sequence of SEQ ID NO: 306, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 308. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 506, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 608. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 504, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 606.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 77, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 315. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 73, a CDR2 comprising the amino acid sequence of SEQ ID NO: 75, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 77, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 311, a CDR2 comprising the amino acid sequence of SEQ ID NO: 313, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 315. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 509, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 611. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 507, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 609.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 84, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 322. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 80, a CDR2 comprising the amino acid sequence of SEQ ID NO: 82, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 84, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 318, a CDR2 comprising the amino acid sequence of SEQ ID NO: 320, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 322. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 512, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 614. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 510, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 612.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 91, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 329. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 87, a CDR2 comprising the amino acid sequence of SEQ ID NO: 89, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 91, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 325, a CDR2 comprising the amino acid sequence of SEQ ID NO: 327, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 329. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 515, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 617. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 513, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 615.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 98, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 336. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 94, a CDR2 comprising the amino acid sequence of SEQ ID NO: 96, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 98, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 332, a CDR2 comprising the amino acid sequence of SEQ ID NO: 334, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 336. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 518, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 620. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 516, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 618.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 105, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 343. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 101, a CDR2 comprising the amino acid sequence of SEQ ID NO: 103, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 105, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 339, a CDR2 comprising the amino acid sequence of SEQ ID NO: 341, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 343. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 521, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 623. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 519, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 621.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein 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: 350. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 108, a CDR2 comprising the amino acid sequence of SEQ ID NO: 110, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 112, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 346, a CDR2 comprising the amino acid sequence of SEQ ID NO: 348, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 350. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 524, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 626. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 522, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 624.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 119, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 357. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 115, a CDR2 comprising the amino acid sequence of SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 119, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 353, a CDR2 comprising the amino acid sequence of SEQ ID NO: 355, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 357. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 527, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 629. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 525, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 627.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein 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: 364. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 122, a CDR2 comprising the amino acid sequence of SEQ ID NO: 124, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 126, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 360, a CDR2 comprising the amino acid sequence of SEQ ID NO: 362, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 364. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 530, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 632. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 528, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 630.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 133, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 371. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 129, a CDR2 comprising the amino acid sequence of SEQ ID NO: 131, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 133, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 367, a CDR2 comprising the amino acid sequence of SEQ ID NO: 369, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 371. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 533, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 635. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 531, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 633.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein 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: 378. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 136, a CDR2 comprising the amino acid sequence of SEQ ID NO: 138, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 140, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 374, a CDR2 comprising the amino acid sequence of SEQ ID NO: 376, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 378. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 536, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 638. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 534, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 636.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 147, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 385. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 143, a CDR2 comprising the amino acid sequence of SEQ ID NO: 145, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 147, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 381, a CDR2 comprising the amino acid sequence of SEQ ID NO: 383, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 385. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 539, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 641. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 537, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 639.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein 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: 392. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 150, a CDR2 comprising the amino acid sequence of SEQ ID NO: 152, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 154, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 388, a CDR2 comprising the amino acid sequence of SEQ ID NO: 390, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 392. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 542, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 644. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 540, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 642.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 161, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 399. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 157, a CDR2 comprising the amino acid sequence of SEQ ID NO: 159, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 161, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 395, a CDR2 comprising the amino acid sequence of SEQ ID NO: 397, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 399. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 545, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 647. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 543, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 645.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein 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: 406. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 164, a CDR2 comprising the amino acid sequence of SEQ ID NO: 166, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 168, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 402, a CDR2 comprising the amino acid sequence of SEQ ID NO: 404, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 406. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 548, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 650. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 546, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 648.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 175, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 413. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 171, a CDR2 comprising the amino acid sequence of SEQ ID NO: 173, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 175, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 409, a CDR2 comprising the amino acid sequence of SEQ ID NO: 411, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 413. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 551, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 653. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 549, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 651.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein, the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 182, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 420. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 178, a CDR2 comprising the amino acid sequence of SEQ ID NO: 180, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 182, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 416, a CDR2 comprising the amino acid sequence of SEQ ID NO: 418, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 420. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 554, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 656. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 552, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 654.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 189, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 427. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 185, a CDR2 comprising the amino acid sequence of SEQ ID NO: 187, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 189, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 423, a CDR2 comprising the amino acid sequence of SEQ ID NO: 425, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 427. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 557, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 659. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 555, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 657.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein, the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 196, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 434. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 192, a CDR2 comprising the amino acid sequence of SEQ ID NO: 194, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 196, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 430, a CDR2 comprising the amino acid sequence of SEQ ID NO: 432, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 434. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 560, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 662. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 558, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 660.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 203, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 441. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 199, a CDR2 comprising the amino acid sequence of SEQ ID NO: 201, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 203, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 437, a CDR2 comprising the amino acid sequence of SEQ ID NO: 439, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 441. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 563, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 665. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 561, and the beta chain of comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 663.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein 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: 448. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 206, a CDR2 comprising the amino acid sequence of SEQ ID NO: 208, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 210, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 444, a CDR2 comprising the amino acid sequence of SEQ ID NO: 446, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 448. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 566, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 668. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 564, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 666.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein 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: 455. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 213, a CDR2 comprising the amino acid sequence of SEQ ID NO: 215, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 217, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 451, a CDR2 comprising the amino acid sequence of SEQ ID NO: 453, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 455. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 569, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 671. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 567, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 669.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 224, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 462. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 220, a CDR2 comprising the amino acid sequence of SEQ ID NO: 222, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 224, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 458, a CDR2 comprising the amino acid sequence of SEQ ID NO: 460, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 462. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 572, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 674. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 570, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 672.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 231, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 469. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 227, a CDR2 comprising the amino acid sequence of SEQ ID NO: 229, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 231, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 465, a CDR2 comprising the amino acid sequence of SEQ ID NO: 467, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 469. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 575, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 677. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 573, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 675.
Described herein is a TCR comprising an alpha chain and a beta chain, wherein, the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 238, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 476. Also described herein is a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 234, a CDR2 comprising the amino acid sequence of SEQ ID NO: 236, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 238, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 472, a CDR2 comprising the amino acid sequence of SEQ ID NO: 474, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 476. In certain embodiments, the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 578, and the beta chain comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 680. In certain embodiments, the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 576, and the beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 678.
The disclosure also provides polynucleotides that encode any of the disclosed TCRs.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 5. Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by a nucleic acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR3 that is encoded by a corresponding nucleic acid sequence provided in Table 5 (i.e. the beta chain CDR3 provided in Table 5 is in the same row number (row 1, for example) as the alpha chain CDR3 provided in Table 4 (row 1, for example)).
Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence provided in Table 4, a CDR2 comprising an amino acid sequence provided in Table 4, and a CDR3 comprising an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 5, a CDR2 comprising an amino acid sequence provided in Table 5, and a CDR3 comprising an amino acid sequence provided in Table 5. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by a nucleic acid sequence provided in Table 4, a CDR2 that is encoded by a nucleic acid sequence provided in Table 4, and a CDR3 that is encoded by a nucleic acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR1 that is encoded by a nucleic acid sequence provided in Table 5, a CDR2 that is encoded by a nucleic acid sequence provided in Table 5, and a CDR3 that is encoded by a nucleic acid sequence provided in Table 5 (i.e. the beta chain CDRs provided in Table 5 are in the same row number (row 1, for example) as the alpha chain CDRs provided in Table 4 (row 1, for example)).
Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a VDJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid sequence provided in Table 4, and the corresponding beta chain comprises a VDJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid sequence provided in Table 5 (i.e. the beta chain VDJ region provided in Table 5 is in the same row number (row 1, for example) as the alpha chain VJ region provided in Table 4 (row 1, for example)).
Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid sequence provided in Table 4, and the corresponding beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid sequence provided in Table 5 (i.e. the beta chain provided in Table 5 is in the same row number (row 1, for example) as the alpha chain provided in Table 4 (row 1, for example)).
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 6, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 244. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 2, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 4, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 6, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 240, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 242, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 244. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 478, and the beta chain comprises a variable, diversity and joining (VDJ) region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 580. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 1, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 239.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 13, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 251. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 9, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 11, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 13, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 247, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 249, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 251. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 481, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 583. In certain embodiments, the alpha chain of is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 8, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 246.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 20, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 258. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 16, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 18, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 20, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 254, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 256, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 258. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 484, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 586. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 15, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 253.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 27, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 265. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 23, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 25, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 27, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 261, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 263, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 265. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 487, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 589. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 22, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 260.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 34, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 272. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 30, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 32, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 34, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 268, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 270, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 272. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 490, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 592. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 29, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 267.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 41, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 279. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 37, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 39, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 41, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 275, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 277, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 279. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 493, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 595. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 36, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 274.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 48, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 286. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 44, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 46, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 48, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 282, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 284, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 286. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 496, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 598. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 43, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 281.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 55, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 293. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 51, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 53, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 55, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 289, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 291, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 293. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 499, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 601. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 50, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 288.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 62, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 300. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 58, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 60, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 62, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 296, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 298, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 300. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 502, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 604. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 57, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 295.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 69, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 307. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 65, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 67, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 69, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 303, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 305, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 307. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 505, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 607. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 64, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 302.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 76, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 314. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 72, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 74, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 76, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 310, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 312, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 314. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 508, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 610. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 71, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 309.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 83, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 321. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 79, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 81, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 83, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 317, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 319, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 321. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 511, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 613. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 78, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 316.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 90, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 328. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 86, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 88, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 90, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 324, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 326, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 328. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 514, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 616. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 85, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 323.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 97, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 335. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 93, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 95, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 97, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 331, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 333, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 335. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 517, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 619. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 92, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 330.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 104, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 342. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 100, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 102, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 104, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 338, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 340, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 342. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 520, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 622. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 99, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 337.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 111, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 349. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 107, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 109, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 111, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 345, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 347, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 349. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 523, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 625. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 106, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 344.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 118, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 356. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 114, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 116, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 118, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 352, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 354, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 356. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 526, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 628. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 113, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 351.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 125, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 363. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 121, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 123, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 125, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 359, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 361, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 363. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 529, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 631. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 120, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 358.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 132, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 370. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 128, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 130, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 132, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 366, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 368, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 370. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 532, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 634. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 127, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 365.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 139, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 377. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 135, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 137, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 139, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 373, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 375, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 377. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 535, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 637. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 134, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 372.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 146, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 384. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 142, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 144, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 146, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 380, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 382, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 384. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 538, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 640. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 141, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 379.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 153, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 391. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 149, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 151, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 153, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 387, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 389, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 391. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 541, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 643. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 148, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 386.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 160, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 398. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 156, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 158, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 160, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 394, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 396, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 398. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 544, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 646. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 155, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 393.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 167, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 405. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 163, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 165, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 167, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 401, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 403, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 405. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 547, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 649. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 162, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 400.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 174, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 412. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 170, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 172, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 174, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 408, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 410, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 412. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 550, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 652. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 169, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 407.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein, the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 181, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 419. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 177, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 179, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 181, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 415, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 417, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 419. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 553, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 655. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 176, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 414.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 188, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 426. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 184, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 186, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 188, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 422, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 424, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 426. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 556, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 658. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 183, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 421.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 195, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 433. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 191, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 193, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 195, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 429, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 431, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 433. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 559, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 661. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 190, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 428.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 202, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 440. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 198, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 200, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 202, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 436, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 438, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 440. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 562, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 664. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 197, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 435.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 209, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 447. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 205, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 207, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 209, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 443, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 445, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 447. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 565, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 667. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 204, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 442.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 216, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 454. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 212, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 214, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 216, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 450, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 452, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 454. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 568, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 670. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 211, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 449.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 223, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 461. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 219, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 221, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 223, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 457, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 459, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 461. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 571, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 673. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 218, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 456.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 230, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 468. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 226, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 228, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 230, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 464, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 466, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 468. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 574, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 676. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 225, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 463.
Described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 237, and the beta chain comprises a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 475. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 233, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 235, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 237, and the beta chain comprises a CDR1 that is encoded by the nucleic acid sequence of SEQ ID NO: 471, a CDR2 that is encoded by the nucleic acid sequence of SEQ ID NO: 473, and a CDR3 that is encoded by the nucleic acid sequence of SEQ ID NO: 475. In certain embodiments, the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 577, and the beta chain comprises a VJD region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 679. In certain embodiments, the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 232, and the beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequence of SEQ ID NO: 470.
In some embodiments, the polynucleotide comprises DNA.
In some embodiments, the polynucleotide comprises RNA.
In some embodiments, the 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.
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.
Described herein are vectors comprising polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 5. Described herein are vectors comprising polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by a nucleic acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR3 that is encoded by a corresponding nucleic acid sequence provided in Table 5.
Also described herein are vectors comprising polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence provided in Table 4, a CDR2 comprising an amino acid sequence provided in Table 4, and a CDR3 comprising an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 5, a CDR2 comprising an amino acid sequence provided in Table 5, and a CDR3 comprising an amino acid sequence provided in Table 5.
Also described herein are vectors comprising polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by a nucleic acid sequence provided in Table 4, a CDR2 that is encoded by a nucleic acid sequence provided in Table 4, and a CDR3 that is encoded by a nucleic acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR1 that is encoded by a nucleic acid sequence provided in Table 5, a CDR2 that is encoded by a nucleic acid sequence provided in Table 5, and a CDR3 that is encoded by a nucleic acid sequence provided in Table 5.
Also described herein are vectors comprising polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a VDJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5.
Also described herein are vectors comprising polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid sequence provided in Table 4, and the corresponding beta chain comprises a VDJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid sequence provided in Table 5.
Also described herein are vectors comprising polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5.
Also described herein are vectors comprising polynucleotides that encode a TCR polypeptide comprising an alpha chain and a beta chain, wherein the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid sequence provided in Table 4, and the corresponding beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid sequence provided in Table 5.
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 include, but are not limited to—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.
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 TCR 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 (VEEV), 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, LNP encapsulated self-replicating RNA derived from alpha virus, 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. Suitable adenovirus vectors include, for example, 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.
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 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 contains deletions 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 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 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 293E16 (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).
The disclosure also provides a host cell comprising any of the above vectors.
Suitable host cells include prokaryotic or eukaryotic host cells. 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 Agel cell line.
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 efficacy or therapeutic impact of a treatment.
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 polynucleotides encoding the polypeptides of the disclosure) by replacing viral sequences encoding structural genes.
Provided herein is a viral vector comprising any of the polynucleotides of the disclosure, wherein the vector is a self-replicating RNA molecule.
Any of the above self-replicating RNA molecules can further comprise one or more of the following:
In some embodiments, for example, the self-replicating RNA molecule can comprise one or more of the following:
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), where the coding sequence for the autoprotease is optionally operably linked upstream to the nucleic acid sequence encoding the GOI.
Generally, any proteolytic cleavage site known in the art can be incorporated into the nucleic acid molecules of the disclosure and can be, for example, proteolytic cleavage sequences that are cleaved post-production by a protease. Further suitable proteolytic cleavage sites also include proteolytic cleavage sequences that can be cleaved following addition of an external protease. As used herein the term “autoprotease” refers to a “self-cleaving” peptide that possesses autoproteolytic activity and is capable of cleaving itself from a larger polypeptide moiety. First identified in the foot-and-mouth disease virus (FMDV), a member of the picornavirus group, several autoproteases have been subsequently identified such as, for example, “2A like” peptides from equine rhinitis A virus (E2A), porcine teschovirus-1 (P2A) and Thosea asigna virus (T2A), and their activities in proteolytic cleavage have been shown in various ex vitro and in vivo eukaryotic systems. As such, the concept of autoproteases is available to one of skill in the art as many naturally occurring autoprotease systems have been identified. Well studied autoprotease systems are e.g. viral proteases, developmental proteins (e.g. HetR, Hedgehog proteins), RumA autoprotease domain, UmuD, etc.). Non-limiting examples of autoprotease peptides suitable for the compositions and methods of the present disclosure include the peptide sequences from 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), or a combination thereof.
In some embodiments, the coding sequence for the autoprotease peptide is operably linked downstream of the DLP motif and upstream to the GOI.
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).
In some embodiments, the autoprotease peptide is selected from the group consisting of porcine teschovirus-1 2A (P2A), foot-and-mouth disease virus (FMDV) 2A (F2A), Equine Rhinitis A Virus (ERAV) 2A (E2A), Thosea asigna virus 2A (T2A), cytoplasmic polyhedrosis virus 2A (BmCPV2A), Flacherie Virus 2A (BmIFV2A), and a combination thereof.
In some embodiments, the autoprotease peptide is porcine teschovirus-1 2A (P2A).
The incorporation of the P2A peptide in the modified viral RNA replicons allows release of protein encoded by GOI (e.g. the polynucleotides encoding the TCR polypeptides of the disclosure) from the capsid-GOI fusion.
In some embodiments disclosed herein, the porcine teschovirus-1 2A (P2A) peptide sequence is engineered in-frame immediately after the DLP sequence and in-frame immediately upstream of all GOI.
Any of the above-disclosed self-replicating RNA molecules can further include a coding sequence downstream Loop (DLP) motif.
Some viruses have sequences capable of forming one or more stem-loop structures which regulate, for example increase, capsid gene expression. Viral capsid enhancer as used herein refers to a regulatory element comprising sequences capable of forming such stem-loop structures. In some examples, the stem-loop structures are formed by sequences within the coding sequence of a capsid protein and named Downstream Loop (DLP) sequence. As disclosed herein, these stem-loop structures or variants thereof can be used to regulate, for example increase, expression level of genes of interest. For example, these stem-loop structures or variants thereof can be used in a recombinant vector (e.g., in a heterologous viral genome) for enhancing transcription and/or translation of coding sequence operably linked downstream thereto.
Alphavirus replication in host cells is known to induce the double-stranded RNA-dependent protein kinase (PKR). PKR phosphorylates the eukaryotic translation initiation factor 2α (eIF2α). Phosphorylation of eIF2α blocks translation initiation of mRNA and in doing so keeps viruses from a completing a productive replication cycle. Infection of cells with Sindbis virus induces PKR that results in phosphorylation of eIF2α, yet the viral subgenomic mRNA is efficiently translated while translation of all other cellular mRNAs is restricted. The efficient translation of the viral subgenomic mRNA in Sindbis virus is made possible by the presence of a stable RNA hairpin loop (or DLP motif) located downstream of the wild type AUG initiator codon for the virus capsid protein (e.g., capsid enhancer). It has been reported that the DLP structure can stall a ribosome on the wild type AUG and this supports translation of the subgenomic mRNA without the requirement for functional eIF2α. Thus, subgenomic mRNAs of Sindbis virus (SINV) as well as of other alphaviruses are efficiently translated even in cells that have highly active PKR resulting in complete phosphorylation of eIF2α.
The DLP structure was first characterized in Sindbis virus (SINV) 26S mRNA and also detected in Semliki Forest virus (SFV). Similar DLP structures have been reported to be present in at least 14 other members of the Alphavirus genus including New World (for example, MAYV, UNAV, EEEV (NA), EEEV (SA), AURAV) and Old World (SV, SFV, BEBV, RRV, SAG, GETV, MIDV, CHIKV, and ONNV) members. The predicted structures of these Alphavirus 26S mRNAs were constructed based on SHAPE (selective 2′-hydroxyl acylation and primer extension) data (Toribio et al., Nucleic Acids Res. May 19; 44(9):4368-80, 2016), the content of which is hereby incorporated by reference). Stable stem-loop structures were detected in all cases except for CHIKV and ONNV, whereas MAYV and EEEV showed DLPs of lower stability (Toribio et al., 2016 supra). The highest DLP activities were reported for those Alphaviruses that contained the most stable DLP structures.
As an example, members of the Alphavirus genus can resist the activation of antiviral RNA-activated protein kinase (PKR) by means of the downstream loop (DLP) present within viral 26S transcripts, which allows an eIF2-independent translation initiation of these mRNAs. The downstream loop (DLP), is located downstream from the AUG in SINV 26S mRNA and in other members of the Alphavirus genus.
In some embodiments, the nucleic acid molecules of the disclosure can include a coding sequence for a GOI operably linked to DLP motif(s) and/or the coding sequence for the DLP motifs.
In some embodiments, the self-replicating RNA molecule of the disclosure comprises a downstream loop (DLP).
In some embodiments, the downstream loop (DLP) comprises at least one RNA-stem-loop.
In some instances, DLP activity depends on the distance between the DLP motif and the initiation codon AUG (AUGi). The AUG-DLP spacing in Alphavirus 26S mRNAs is tuned to the topology of the ES6S region of the ribosomal 18S rRNA in a way that allows the placement of the AUGi in the P site of the 40S subunit stalled by the DLP, allowing the incorporation of Met-tRNA without the participation of eIF2. In the case of Sindbis virus, the DLP motif is found in the first ˜150 nt of the Sindbis subgenomic RNA. The hairpin is located downstream of the Sindbis capsid AUG initiation codon (AUG at nt 50 of the Sindbis subgenomic RNA) and results in stalling a ribosome such that the correct capsid gene AUG is used to initiate translation. Previous studies of sequence comparisons and structural RNA analysis revealed the evolutionary conservation of DLP in SINV and predicted the existence of equivalent DLP structures in many members of the Alphavirus genus (see e.g., Ventoso, J. Virol. 9484-9494, Vol. 86, September 2012).
Without being bound by any particular theory, it is believed that placing the DLP motif upstream of a coding sequence for any GOI typically results in a fusion-protein of N-terminal capsid amino acids that are encoded in the hairpin region to the GOI encoded protein because initiation occurs on the capsid AUG not the GOI AUG.
In some embodiments, the self-replicating RNA molecule comprises a downstream loop placed upstream of the non-structural protein 1 (nsP1).
In some embodiments, the downstream loop is placed upstream of the non-structural protein 1 (nsP1) and is joined to the nsP1 by a porcine teschovirus-1 2A (P2A) ribosome skipping element.
The DLP-containing self-replicating RNA of the disclosure can be useful in conferring a resistance to the innate immune system in a subject. Unmodified RNA replicons are sensitive to the initial innate immune system state of cells they are introduced into. If the cells/individuals are in a highly active innate immune system state, the RNA replicon performance (e.g., replication and expression of a GOI) can be negatively impacted. By engineering a DLP to control initiation of protein translation, particularly of non-structural proteins, the impact of the pre-existing activation state of the innate immune system to influence efficient RNA replicon replication is removed or lessened. The result is more uniform and/or enhanced expression of a GOI that can impact efficacy or therapeutic impact of a treatment.
The DLP motif of the self-replicating RNA can confer efficient mRNA translation in cellular environments where cellular mRNA translation is inhibited. When a DLP is linked with translation of a replicon vector's non-structural protein genes the replicase and transcriptase proteins are capable of initiating functional replication in PKR activated cellular environments. When a DLP is linked with translation of subgenomic mRNAs robust GOI expression is possible even when cellular mRNA is restricted due to innate immune activation. Accordingly, engineering self-replicating RNA that contain DLP structures to help drive translation of both non-structural protein genes and subgenomic mRNAs provides a powerful way to overcome innate immune activation.
Examples of a self-replicating RNA vector comprising a DLP motif are described in US Patent Application Publication US2018/0171340 and the International Patent Application Publication WO2018106615, the content of which is incorporated herein by reference in its entirety.
Any of the above-disclosed self-replicating RNA molecules can further comprise nonstructural genes nsP1, nsP2, nsP3 and/or nsP4. In some embodiments, the self-replicating RNA molecule does not encode a functional viral structural protein.
Alphavirus genomes encode non-structural proteins nsP1, nsP2, nsP3, and nsP4, which are produced as a single polyprotein precursor, sometimes designated P1234 (or nsP1-4 or nsP1234), and which is cleaved into the mature proteins through proteolytic processing. nsP1 can be about 60 kDa in size and may have methyltransferase activity and be involved in the viral capping reaction. nsP2 has a size of about 90 kDa and may have helicase and protease activity while nsP3 is about 60 kDa and contains three domains: a macrodomain, a central (or alphavirus unique) domain, and a hypervariable domain (HVD). nsP4 is about 70 kDa in size and contains the core RNA-dependent RNA polymerase (RdRp) catalytic domain. After infection the alphavirus genomic RNA is translated to yield a P1234 polyprotein, which is cleaved into the individual proteins.
Alphavirus genomes also encode three structural proteins: the core nucleocapsid protein C, and the envelope proteins P62 and E1 that associate as a heterodimer. Structural proteins are under the control of a subgenomic promoter and can be replaced by the GIO.
In some embodiments, the self-replicating RNA can lack (or not contain) the sequence(s) of at least one (or all) of the structural viral proteins (e.g. nucleocapsid protein C, and envelope proteins P62, 6K, and E1). In these embodiments, the sequences encoding one or more structural genes can be substituted with one or more sequences such as, for example, a coding sequence for at least one protein or peptide (or other gene of interest (GOI)) e.g. the TCR polypeptides of the disclosure.
In some embodiments, the self-replicating RNA lack sequences encoding alphavirus structural proteins; or do not encode alphavirus (or, optionally, any other) structural proteins. In some embodiments, the self-replicating RNA molecules are further devoid of a part or the entire coding region for one or more viral structural proteins. For example, the alphavirus expression system may be devoid of a portion of or the entire coding sequence for one or more of the viral capsid protein C, E1 glycoprotein, E2 glycoprotein, E3 protein and 6K protein.
In some embodiments, the self-replicating RNA molecule does not contain coding sequences for at least one of the structural viral proteins. In these instances, the sequences encoding structural genes can be substituted with one or more sequences such as, for example, a coding sequence for a GOI (e.g., the TCR polypeptides).
The disclosure also provides a self-replicating RNA molecule comprising nonstructural genes nsP1, nsP2, nsP3 and nsP4, and wherein the self-replicating RNA molecule does not encode a functional viral structural protein.
In some embodiments, the disclosure provides a self-replicating RNA molecule comprising the coding sequence for at least one, at least two, at least three, or at least four nonstructural viral proteins (e.g. nsP1, nsP2, nsP3, nsP4). The nsP1, nsP2, nsP3, and nsP4 proteins encoded by the replicon are functional or biologically active proteins.
In some embodiments, the self-replicating RNA molecule includes the coding sequence for a portion of the at least one nonstructural viral protein. For example, the self-replicating RNA molecules can include about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 100%, or a range between any two of these values, of the encoding sequence for the at least one nonstructural viral protein. In some embodiments, the self-replicating RNA molecule can include the coding sequence for a substantial portion of the at least one nonstructural viral protein. As used herein, a “substantial portion” of a nucleic acid sequence encoding a nonstructural viral protein comprises enough of the nucleic acid sequence encoding the nonstructural viral protein to afford putative identification of that protein, either by manual evaluation of the sequence by one skilled in the art, or by computer-automated sequence comparison and identification using algorithms such as BLAST (see, for example, in “Basic Local Alignment Search Tool”; Altschul S F et al., J. Mol. Biol. 215:403-410, 1993).
In some embodiments, the self-replicating RNA molecule can include the entire coding sequence for the at least one nonstructural protein. In some embodiments, the self-replicating RNA molecule comprises substantially all the coding sequence for the native viral nonstructural proteins. In certain embodiments, the one or more nonstructural viral proteins are derived from the same virus.
In some embodiments, the downstream loop DLP of the self-replicating RNA molecule placed upstream of the non-structural protein 1 (nsP1) is derived from Sindbis virus.
In some embodiments, the self-replicating RNA molecule comprises nsP1, nsP2, nsP3, and nsP4 sequences derived from the Venezuelan equine encephalitis virus (VEEV) and a DLP motif derived from the Sindbis virus (SIN).
In some embodiments, the self-replicating RNA molecules also have an RNA sub-sequence encoding an amino acid sequence derived from an alphavirus nsP3 macro domain, and an RNA sub-sequence encoding an amino acid sequence derived from an alphavirus nsP3 central domain. The self-replicating RNA molecules can also have an RNA sub-sequence encoding an amino acid sequence derived entirely from an Old World alphavirus nsP3 hypervariable domain, or can have an amino acid sequence having a portion derived from a New World alphavirus nsP3 hypervariable domain and a portion derived from an Old World alphavirus nsP3 hypervariable domain. i.e. the hyper variable domain (HVD) can be a hybrid or chimeric New World/Old World sequence.
In some embodiments, the self-replicating RNA molecules can have an RNA sequence encoding amino acid sequences derived from wild type New World alphavirus nsP1, nsP2, nsP3, and nsP4 protein sequences. In other embodiments, the one or more nonstructural proteins are derived from different viruses.
In some embodiments, the self-replicating RNA molecule may have an RNA sequence encoding an nsP3 macro domain derived from a wild type alphavirus nsP3, and an nsP3 central domain derived from a wild type alphavirus nsP3. In various embodiments, the macro and central domain(s) can both be derived from a New World wild type alphavirus nsP3 or can both be derived from an Old World wild type alphavirus nsP3 protein. In other embodiments, the macro domain can be derived from a New World wild type alphavirus macro domain and the central domain can be derived from an Old World wild type alphavirus central domain, or vice versa. The various domains can be of any sequence described herein.
In some embodiments, the self-replicating RNA molecule contains non VEEV nonstructural proteins nsP1, nsP2, nsP3 and nsP4.
The accumulated experimental evidence has demonstrated that replication/amplification of VEEV and other alphavirus genomes and their defective interfering (DI) RNAs is determined by three promoter elements: (i) the conserved 3′-terminal sequence element (3′ CSE) and the following poly(A) tail; (ii) the 5′ UTR, which functions as a key promoter element for both negative- and positive-strand RNA synthesis; and (iii) the 51-nt conserved sequence element (51-nt CSE), which is located in the nsP1-coding sequence and functions as an enhancer of alphavirus genome replication (Kim et al., PNAS, 2014, 111: 10708-10713, and references therein).
Any of the above-disclosed self-replicating RNA molecules can further include an unmodified 5′ untranslated region (5′UTR).
Previous studies have demonstrated that during VEEV and Sindbis virus infections only a small portion of viral nonstructural proteins (nsPs) is colocalized with dsRNA replication intermediates. Thus, it appears that a large fraction of nsPs are not involved in RNA replication (Gorchakov R, et al. (2008) A new role for ns polyprotein cleavage in Sindbis virus replication. J Virol 82(13):6218-6231). This has provided an opportunity to exploit the under used ns proteins for amplification of the subgenomic RNAs encoding proteins of interest, which is normally transcribed from the subgenomic promoter and is not further amplified.
In some embodiments, a fragment of the nsP1 of the self-replicating RNA molecule of the disclosure is duplicated downstream of the 5′-UTR and upstream of the DLP.
In some embodiments, the first 193 nucleotides of nsP1 are duplicated downstream of the 5′ UTR and upstream of the DLP.
In some embodiments, the self-replicating RNA molecule comprises a modified 5′ untranslated region (5′-UTR). For example, the modified 5′-UTR can comprise one or more nucleotide substitutions at position 1, 2, 4, or a combination thereof. Preferably, the modified 5′-UTR comprises a nucleotide substitution at position 2, more preferably the modified 5′-UTR has a U->G substitution at position 2. Examples of such self-replicating RNA molecules are described in US Patent Application Publication US2018/0104359 and International Patent Application Publication WO2018075235, the content of which is incorporated herein by reference in its entirety.
In some embodiments, the UTRs can be wild type New World or Old World alphavirus UTR sequences, or a sequence derived from any of them. The 5′ UTR can be of any suitable length, such as about 60 nt or 50-70 nt or 40-80 nt. In some embodiments the 5′ UTR can also have conserved primary or secondary structures (e.g. one or more stem-loop(s)) and can participate in the replication of alphavirus or of replicon RNA. The 3′ UTR can be up to several hundred nucleotides, for example it can be 50-900 or 100-900 or 50-800 or 100-700 or 200 nt-700 nt. The ′3 UTR also can have secondary structures, e.g. a step loop, and can be followed by a polyadenylate tract or poly-A tail.
The 5′ and 3′ untranslated regions can be operably linked to any of the other sequences encoded by the replicon. The UTRs can be operably linked to a promoter and/or sequence encoding a protein or peptide by providing sequences and spacing necessary for recognition and transcription of the other encoded sequences.
The GOI (e.g. the polynucleotides encoding the TCR polypeptides of the disclosure) can be expressed under the control of a subgenomic promoter. In certain embodiments, instead of the native subgenomic promoter, the subgenomic RNA can be placed under control of internal ribosome entry site (IRES) derived from encephalomyocarditis viruses (EMCV), Bovine Viral Diarrhea Viruses (BVDV), polioviruses, Foot-and-mouth disease viruses (FMD), enterovirus 71, or hepatitis C viruses. Subgenomic promoters range from 24 nucleotides (Sindbis virus) to over 100 nucleotides (Beet necrotic yellow vein virus) and are usually found upstream of the transcription start.
The self-replicating RNA molecules can have a 3′ poly-A tail. It can also include a poly-A polymerase recognition sequence (e.g. AAUAAA) near its 3′ end.
In those instances where the self-replicating RNA molecule is to be packaged into a recombinant alphavirus particle, it can contain one or more sequences, so-called packaging signals, which serve to initiate interactions with alphavirus structural proteins that lead to particle formation. In some embodiments, the alphavirus particles comprise RNA derived from one or more alphaviruses, and structural proteins wherein at least one of said structural proteins is derived from two or more alphaviruses.
In some embodiments, the self-replicating RNA molecule comprises a VEEV derived vector wherein the structural viral proteins (e.g. nucleocapsid protein C, and envelope proteins P62, 6K, and E1) are removed and replaced by the coding sequence of the polypeptides of the disclosure.
Engineered cells expressing the TCRs that specifically bind the CALR and JAK2 epitopes of the disclosure are within the scope of the disclosure. The present disclosure provides engineered cells (e.g., T cells) comprising any one of an alpha TCR chain and any one of a beta TCR chain, as disclosed in Tables 4 and 5. These engineered cells can be used to treat various disorders or disease, such as cancer, myeloproliferative disease and cardiovascular disease.
Suitable engineered cells include those obtained from, for example, animals and humans.
In some embodiments, the cell comprising the TCR of the disclosure is a T cell, a natural killer (NK) cell, a cytotoxic T lymphocyte (CTL), a regulatory T cell (Treg), a human embryonic stem cell, a lymphoid progenitor cell, a T cell-precursor cell, a pluripotent stem cell or induced pluripotent stem cell (iPSC) from which lymphoid cells may be differentiated. In some embodiments, the isolated cell comprising the TCR of the disclosure is a T cell. The T cell may be any T cell, such as a cultured T cell, e.g., a primary T cell, or a T cell from a cultured T cell line, e.g., Jurkat, SupT1, etc., or a T cell obtained from a mammal. If obtained from a mammal, the T cell can be obtained from any source, including bone marrow, blood, lymph node, thymus, or other tissues or fluids. T cells may also be enriched for or purified. The T cell may be a human T cell. The T cell may be a T cell isolated from a human. The T cell can be any type of T cell and may be of any developmental stage, including, CD4+CD8+ double positive T cells, CD8+ T cells (e.g., cytotoxic T cells), CD4+ helper T cells, e.g., Th1 and Th2 cells, peripheral blood mononuclear cells (PBMCs), peripheral blood leukocytes (PBLs), tumor infiltrating cells, memory T cells, naïve T cells, and the like. The T cell may be a CD8+ T cell or a CD4+ T cell. The T cell may be an αβ T cell or a γδ T cell.
In some embodiments, the isolated cell comprising the TCR of the disclosure is a NK cell.
In some embodiments, the isolated cell comprising the TCR of the disclosure is a αβ T cell.
In some embodiments, the isolated cell comprising the TCR of the disclosure is a γδ T cell.
In some embodiments, the isolated cell comprising the TCR of the disclosure is a CD8+ cytotoxic T lymphocyte
In some embodiments, the isolated cell comprising the TCR of the disclosure is a human embryonic stem cell.
In some embodiments, the isolated cell comprising the TCR of the disclosure is a lymphoid progenitor cell.
In some embodiments, the isolated cell comprising the TCR of the disclosure is a pluripotent stem cell.
In some embodiments, the isolated cell comprising the TCR of the disclosure is an induced pluripotent stem cell (iPSC).
In some embodiments, the isolated cell comprising the TCR of the disclosure is a CD4+T lymphocyte. In certain embodiments, the CD4+T lymphocyte is a Th1 cell. In certain embodiments, the CD4+T lymphocyte is a Th2 cell. In certain embodiments, the CD4+T lymphocyte is a Th17 cell. In certain embodiments, the CD4+T lymphocyte is a Treg cell. In certain embodiments, the CD4+T lymphocyte is a TFH cell
Described herein are methods for generating engineered cells comprising any one of an alpha TCR chain and any one of a beta TCR chain, as disclosed in Tables 4 and 5.
In some embodiments, to produce an engineered cell for use as a cell therapy product, cells, preferably autologous T cells, are obtained from a pheresis sample withdrawn from a subject and are modified with a vector of the disclosure that encodes the desired TCR alpha and beta sequences. When encountered by the engineered cells, targeted cells may be killed by the engineered cells by virtue of the ability of the engineered cells, such as engineered T cells, to exhibit specific target cell cytotoxicity (i.e., specific cell killing).
The pheresis sample may be collected from the subject by any of a number of suitable lymphocytapheresis, lymphapheresis, and leukaphoresis procedures now known or that become available in the art, which provide for the collection of peripheral blood leukocytes (PBLs) from collected peripheral blood, and from which leukocytes may be separated from other plasma components of the sample. Exemplary procedures are illustrated in, e.g., U.S. Patent Application Publication Numbers US 2004/0173778 and US 200/40147021, and U.S. Pat. Nos. 4,690,915, 5,126,132, 6,255,073, 5,846,827, 6,251,385, 6,225,044, 6,210,963, 6,194,207, 5,443,983, 6,040,177, 5,766,920, 6,210,662, 6,204,058, and 6,227,368.
Selected naive cells, preferably naïve T cells, may be substantially separated from non-selected pheresis sample components. In certain embodiments, naive T cells, which may be naive CD4+ T cells, naive CD8+ T cells, or naive CD4+ T cells and naive CD8+ T cells, are substantially separated from other PBLs, e.g., non-T cells. Methods for selection of PBLs include procedures employing Ficoll gradients, technique employing immunopurification (e.g., monoclonal antibodies directed against cell surface markers, such as CD molecules, and beads, such as Sepharose-, Protein A-, and Protein G-conjugated beads to which the antibodies may be adsorbed, and magnetic beads to which antibodies may be adsorbed), flow cytometry, and fluorescence-activated cell sorter (FACS) analysis.
In certain embodiments, selected naive T cells are substantially purified by magnetic bead purification systems such as those available in the art, e.g., Miltenyi beads (Myltenyi Biotec) and Dynabead systems (Dynal Biotech) combined with cell sorting procedures, such as FACS-based methods, or other appropriate cell sorting devices and methodologies.
A subject from whom a pheresis product comprising naive cells may be obtained is preferably a mammal in need of treatment, such as a dog, a cat, a horse, a rat, rabbit, mouse, a non-human primate, or a human. More preferably, the subject is a human patient in need of treatment for a cancer, a myeloproliferative disease, or a cardiovascular disease. Alternatively, in appropriate circumstances immune cells, such as naive T cells, that are not derived from a subject to be treated, but which are derived from another compatible source such as an immune cell donor, or even an immortalized or transformed immune cell line, may be employed to prepare cell therapy products in accordance with the invention.
The thus-selected naive cells, preferably naïve T cells, are then modified introducing a viral vector comprising any of the disclosed polynucleotides into the cells using known methods. The cells are able to replicate in vivo, resulting in long-term persistence that can lead to sustained tumor control.
In some embodiments, the naive cells, preferably naïve T cells, are transduced with any of the disclosed TCRs, resulting in constitutive expression of the TCR on the surface of the cell. The cells expressing the TCR may further be engineered to express one or more co-stimulatory molecules. Exemplary co-stimulatory molecules are CD28, ICOS, LIGHT, GITR, 4-1BB and OX40. The cells expressing the TCR may further be engineered to produce one or more cytokines or chemokines or proinflammatory mediators, such as TNFα, IFNγ, IL-2, IL-3, IL-6, IL-7, IL-11, IL-12, IL-15, IL-17 or IL-21. The cells may have their endogenous TCR locus and/or HLA locus inactivated using known gene editing technologies.
The engineered cells may optionally be expanded to produce therapeutic compositions and cell therapy products comprising engineered cells of a desired phenotype and number. An exemplary T cell expansion procedure includes incubating activated T cells with irradiated non-CD8+ cells in the presence of selected cytokines and an anti-CD3 antibody preparation to promote non-specific activated T cell expansion. Selection of the expansion protocols to be employed are within the purview of the artisan and may be facilitated by guidance in the art.
The disclosure also provides compositions comprising any of the polynucleotides, any of the polypeptides, or any of the vectors disclosed herein. In some embodiments, the compositions may comprise a vector comprising any of the nucleotides disclosed herein.
The compositions 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.
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.
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 provided herein are also prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.
Provided herein are methods for treating a subject with any of the disclosed compositions or engineered cells. The compositions and engineered cells of the disclosure may be used to treat, prevent, or reduce the risk of a clinical condition.
In some embodiments, the clinical condition is a cancer, a myeloproliferative disease, or a cardiovascular disease.
In some embodiments, the myeloproliferative disease is selected from primary myelofibrosis (MPN), polycythemia vera (PV), essential thrombocythemia (ET), primary myelofibrosis (PFM), secondary myelofibrosis, acute myeloid leukemia (AML), secondary AML, chronic myelogenous leukemia (CML), clonal hematopoiesis of indeterminate potential (CHIP), and chronic myelomonocytic leukemia (CMML).
In some embodiments, the cancer is selected from lung cancer, lymphoid cancer, acute lymphoid leukemia, acute myeloid leukemia, chronic myelogenous leukemia, Burkitt's lymphoma, Hodgkin's lymphoma, plasma cell myeloma, biliary tract cancer, bladder cancer, liver cancer, pancreatic cancer, prostate cancer, skin cancer, thyroid cancer, stomach cancer, large intestine cancer, colon cancer, urinary tract cancer, central nervous system cancer, neuroblastoma, kidney cancer, breast cancer, cervical cancer, testicular cancer, and soft tissue cancer.
In some embodiments, the cardiovascular disease is selected from an acute coronary syndrome, an ischemic cerebrovascular disease, an ischemic heart disease, a thrombosis, a venous thromboembolism, a deep vein thrombosis, a pulmonary embolism, a catastrophic intra-abdominal thromboses, a peripheral arterial disease, a hypertension, a heart failure, an atrial fibrillation, a coronary heart disease, an atherosclerosis, and a clonal hematopoiesis.
In some embodiments, the methods comprise administering to the subject in need thereof a composition comprising any of the polynucleotides, any of the polypeptides, or any of the vectors disclosed herein. In some embodiments, the compositions may comprise a vector comprising any of the nucleotides disclosed herein.
In some embodiments, the methods comprise administering to the subject in need thereof a composition comprising any of the polynucleotides disclosed herein, wherein the polynucleotides encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 5. In some embodiments, the methods disclosed herein comprise administering to the subject in need thereof a composition comprising polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 that is encoded by a nucleic acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR3 that is encoded by a corresponding nucleic acid sequence provided in Table 5 (i.e. the beta chain CDR3 provided in Table 5 is in the same row number (row 1, for example) as the alpha chain CDR3 provided in Table 4 (row 1, for example)). For the sake of brevity, each possible alternative is not parsed out, but each constitute as separate embodiments as if fully described.
In some embodiments, the methods comprise administering to the subject in need thereof a composition comprising polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising the amino acid sequence provided in Table 4, a CDR2 comprising an amino acid sequence provided in Table 4, and a CDR3 comprising an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 5, a CDR2 comprising an amino acid sequence provided in Table 5, and a CDR3 comprising an amino acid sequence provided in Table 5. In some embodiments, the methods comprise administering to the subject in need thereof a composition comprising polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 that is encoded by a nucleic acid sequence provided in Table 4, a CDR2 that is encoded by a nucleic acid sequence provided in Table 4, and a CDR3 that is encoded by a nucleic acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR1 that is encoded by a nucleic acid sequence provided in Table 5, a CDR2 that is encoded by a nucleic acid sequence provided in Table 5, and a CDR3 that is encoded by a nucleic acid sequence provided in Table 5 (i.e. the beta chain CDRs provided in Table 5 are in the same row number (row 1, for example) as the alpha chain CDRs provided in Table 4 (row 1, for example)). For the sake of brevity, each possible alternative is not parsed out, but each constitute separate embodiments as if fully described.
In some embodiments, the methods comprise administering to the subject in need thereof a composition comprising polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a VDJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5. In some embodiments, the methods comprise administering to the subject in need thereof a composition comprising polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a VJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid sequence provided in Table 4, and the corresponding beta chain comprises a VDJ region that is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid sequence provided in Table 5 (i.e. the beta chain VDJ region provided in Table 5 is in the same row number (row 1, for example) as the alpha chain VJ region provided in Table 4 (row 1, for example)). For the sake of brevity, each possible alternative is not parsed out, but each constitute separate embodiments as if fully described.
In some embodiments, the methods comprise administering to the subject in need thereof a composition comprising polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5. Also described herein are polynucleotides that encode a TCR comprising an alpha chain and a beta chain, wherein the alpha chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid sequence provided in Table 4, and the corresponding beta chain is encoded by a nucleic acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to a nucleic acid sequence provided in Table 5 (i.e. the beta chain provided in Table 5 is in the same row number (row 1, for example) as the alpha chain provided in Table 4 (row 1, for example)). For the sake of brevity, each possible alternative is not parsed out, but each constitute separate embodiments as if fully described.
In some embodiments, the methods comprise administering to the subject in need thereof a composition comprising a viral vector comprising a polynucleotide encoding any of the TCR polypeptides of the disclosure. In any of the methods disclosed herein, the composition that is administered to a subject may comprise a vector 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 some embodiments, the vector is selected from Ad26 vector, MVA vector, GAd20 vector, a self-replicating RNA molecule, and combinations thereof.
In some embodiments, the methods comprise administering to the subject in need thereof a composition comprising any of the polypeptides disclosed herein, wherein the polypeptides are TCRs. In some embodiments, the methods comprise administering to the subject in need thereof a composition comprising a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 4, and the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 5. (i.e. the beta chain CDR3 provided in Table 5 is in the same row number (row 1, for example) as the alpha chain CDR3 provided in Table 4 (row 1, for example)). For the sake of brevity, each possible alternative is not parsed out, but each constitute separate embodiments as if fully described.
In some embodiments, the methods comprise administering to the subject in need thereof a composition comprising a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 4, a CDR2 comprising an amino acid sequence provided in Table 4, and a CDR3 comprising an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 5, a CDR2 comprising an amino acid sequence provided in Table 5, and a CDR3 comprising an amino acid sequence provided in Table 5 (i.e. the beta chain CDRs provided in Table 5 are in the same row number (row 1, for example) as the alpha chain CDRs provided in Table 4 (row 1, for example)). For the sake of brevity, each possible alternative is not parsed out, but each constitute separate embodiments as if fully described.
In some embodiments, the methods comprise administering to the subject in need thereof a composition comprising a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a variable and joining (VJ) region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a variable, diversity and joining (VDJ) region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5 (i.e. the beta chain VDJ region provided in Table 5 is in the same row number (row 1, for example) as the alpha chain VJ region provided in Table 4 (row 1, for example)). For the sake of brevity, each possible alternative is not parsed out, but each constitute separate embodiments as if fully described.
In some embodiments, the methods comprise administering to the subject in need thereof a composition comprising a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5 (i.e. the beta chain provided in Table 5 is in the same row number (row 1, for example) as the alpha chain provided in Table 4 (row 1, for example)). For the sake of brevity, each possible alternative is not parsed out, but each constitute separate embodiments as if fully described.
In some embodiments, the methods comprise administering to the subject in need thereof any of the engineered cells disclosed herein. In certain embodiments, the engineered cells comprise any one of an alpha TCR chain and any one of a beta TCR chain, as disclosed in Tables 4 and 5. For the sake of brevity, each possible alternative is not parsed out, but each constitute separate embodiments as if fully described.
In some embodiments, the methods comprise administering to the subject in need thereof an engineered cell comprising any of the polypeptides disclosed herein, wherein the polypeptides are TCRs. In some embodiments, the methods comprise administering to the subject in need thereof an engineered cell comprising a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR3 comprising an amino acid sequence provided in Table 4, and the beta chain comprises a CDR3 comprising a corresponding amino acid sequence provided in Table 5. (i.e. the beta chain CDR3 provided in Table 5 is in the same row number (row 1, for example) as the alpha chain CDR3 provided in Table 4 (row 1, for example)). For the sake of brevity, each possible alternative is not parsed out, but each constitute separate embodiments as if fully described.
In some embodiments, the methods comprise administering to the subject in need thereof an engineered cell comprising a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a CDR1 comprising an amino acid sequence provided in Table 4, a CDR2 comprising an amino acid sequence provided in Table 4, and a CDR3 comprising an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a CDR1 comprising an amino acid sequence provided in Table 5, a CDR2 comprising an amino acid sequence provided in Table 5, and a CDR3 comprising an amino acid sequence provided in Table 5 (i.e. the beta chain CDRs provided in Table 5 are in the same row number (row 1, for example) as the alpha chain CDRs provided in Table 4 (row 1, for example)). For the sake of brevity, each possible alternative is not parsed out, but each constitute separate embodiments as if fully described.
In some embodiments, the methods comprise administering to the subject in need thereof an engineered cell comprising a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises a variable and joining (VJ) region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises a variable, diversity and joining (VDJ) region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5 (i.e. the beta chain VDJ region provided in Table 5 is in the same row number (row 1, for example) as the alpha chain VJ region provided in Table 4 (row 1, for example)). For the sake of brevity, each possible alternative is not parsed out, but each constitute separate embodiments as if fully described.
In some embodiments, the methods comprise administering to the subject in need thereof an engineered cell comprising a TCR comprising an alpha chain and a beta chain, wherein the alpha chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 4, and the corresponding beta chain comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence provided in Table 5 (i.e. the beta chain provided in Table 5 is in the same row number (row 1, for example) as the alpha chain provided in Table 4 (row 1, for example)). For the sake of brevity, each possible alternative is not parsed out, but each constitute separate embodiments as if fully described.
In some embodiments, the engineered cell comprising the TCR is a T cell, a natural killer (NK) cell, a cytotoxic T lymphocyte (CTL), a regulatory T cell (Treg), a human embryonic stem cell, a lymphoid progenitor cell, a T cell-precursor cell, a pluripotent stem cell or induced pluripotent stem cell (iPSC) from which lymphoid cells may be differentiated.
In certain embodiments, the engineered cell is derived from naïve cells that are obtained from a subject in need of treatment. In certain embodiments, the engineered cell is not derived from a subject in need of treatment, but is instead derived from another compatible source such as an immune cell donor, or an immortalized or transformed immune cell line.
In some embodiments, desired amounts of engineered cells are employed to prepare a cell therapy product for therapeutic administration. In certain embodiments, the cell therapy product is reinfused or transfused back into the subject from whom the pheresis sample used to derive the cell therapy product was obtained. Reinfusion procedures that may be employed include those procedures disclosed in, for example, U.S. Pat. Nos. 4,844,893 and 4,690,915. In certain embodiments, the cell therapy product is reinfused or transfused back into a different subject from whom the pheresis sample used to derive the cell therapy product was obtained.
In some embodiments, prior to administration one or more quality assurance tests are performed on the activated T lymphocytes or cell therapy product. In some embodiments, the quality assurance testing comprises performing one or more tests to confirm: HLA match between patient and T lymphocytes; flow cytometry analysis (CD8+, TCR+); sterility (no bacterial or fungal growth); gram-stain negative for bacteria; mycoplasma negative for PCR/ELISA; no residual Drosophila DNA; absence of insect virus cDNA; viability (>72% viable); and cytolytic activity by CTL assay.
In some embodiments, desired amounts of viral vectors are employed to prepare a composition for therapeutic administration.
In some embodiments, the compositions comprising viral vectors is administered at a dose from about 1×104 IFU (Infectious Unit) to about 1×1011 IFU per dose. In certain embodiments, the viral vector is an adenovirus vector, an Ad26 vector, a GAd20 vector, a poxvirus vector or an MVA vector.
In some embodiments, the compositions comprising self-replicating RNA molecule is administered at a dose from about 1 microgram to about 100 micrograms.
In some embodiments, the compositions disclosed herein may be administered to a subject by a variety of routes such as subcutaneous, topical, oral and intramuscular. Administration of the compositions may be accomplished orally or parenterally. Methods of parenteral delivery include topical, intra-arterial (directly to the tissue), intramuscular, subcutaneous, intramedullary, intrathecal, intraventricular, intravenous, intraperitoneal, or intranasal administration. The present disclosure also has the objective of providing suitable topical, oral, systemic and parenteral formulations for use in the methods of prophylaxis and treatment.
In some embodiments, intramuscular administration of the composition can be achieved by using a needle. An alternative is the use of a needleless injection device to administer the composition (using, e.g., Biojector™) or a freeze-dried powder containing the composition. For intravenous, cutaneous or subcutaneous injection, or injection at the site of affliction, the composition may be the form of a parenterally acceptable aqueous solution which is pyrogen-free and has suitable pH, isotonicity and stability. Those of skill in the art are well able to prepare suitable solutions using, for example, isotonic vehicles such as Sodium Chloride Injection, Ringer's Injection, Lactated Ringer's Injection. Preservatives, stabilizers, buffers, antioxidants and/or other additives can be included, as required. A slow-release formulation may also be employed.
The actual amount administered, and rate and time-course of administration, will depend on the nature and severity of what is being treated. Prescription of treatment, e.g., decisions on dosage etc., is within the responsibility of general practitioners and other medical doctors, and typically takes account of the disorder to be treated, the condition of the individual patient, the site of delivery, the method of administration and other factors known to practitioners.
In some embodiments, desired amounts of engineered cells are employed to prepare a cell therapy product for therapeutic administration.
To treat a subject, an effective amount of a cell therapy product is administered to a subject suffering from or diagnosed as having a disease, disorder, or condition. The subject can be a human patient in need of treatment for a cancer, a myeloproliferative disease, or a cardiovascular disease.
Effective amounts or doses of the cell therapy products may be ascertained by routine methods such as modeling, dose escalation studies or clinical trials, and by taking into consideration routine factors, e.g., the mode or route of administration or product delivery, the pharmacokinetics of the cell therapy product, the severity and course of the disease, disorder, or condition, the subject's previous or ongoing therapy, the subject's health status and response to drugs, and the judgment of a treating physician. As exemplary dosage amounts, cell populations may comprise from about 1×106 to about 1×1012 activated T cells, such as 1×108 to 1×1011 or 1×109 to 1×1010 activated T cells for an adult human.
The cell therapy product can be prepared as a therapeutic composition comprising engineered cells, preferably engineered T cells, and a vehicle suitable for the maintenance of the engineered cells until they are infused into the subject, such as a pharmaceutically acceptable diluent or solvent.
Any suitable technique for administering compositions comprising cellular components into a subject may be employed. For example, administration of the engineered T cell via intravenous infusion may be employed. Multiple infusions may be required or indicated, and these infusions may occur over a period of several weeks or longer. Exemplary techniques are described in, for example, U.S. Pat. Nos. 4,844,893 and 4,690,915.
These examples are provided for illustrative purposes only and not to limit the scope of the claims provided herein.
Single cell TCR α/β and RNA sequencing was performed on CD8+ T cell samples isolated from a leukopak from four healthy human donors. Dendritic cells from each donor were treated with the following: a) DMSO treatment or exogenously loaded single 9-mer peptide corresponding to one of two mutJAK2 epitopes (“exogenous treatment”), b) transfection with a tandem minigene construct composed of the combined mutJAK2 and mutCALR epitopes (mutCALR type I and mutCALR type II) (“endogenous treatment”), or c) transduction with an Ad5 virus containing the mutJAK2 and mutCALR antigen construct (“Ad5 treatment”).
The 9-mer peptides corresponding to two mutJAK2 epitopes have the following amino acid sequences: VLNYGVCFC (SEQ ID NO: 681) and FCGDENILV (SEQ ID NO: 682). The tandem minigene construct composed of the combined mutJAK2 and mutCALR epitopes has the amino acid sequence:
In SEQ ID NO: 683, the mutCALR type 1 sequence is underlined, the mutCALR type II sequence is underlined and bold, and the two mutJAK2 sequences are underlined an italics.
The mutJAK2 and mutCALR antigen construct (also known as LS_CALR_JAK2-2x9mer (HEME002)) that is contained within the Ad5 virus has the following amino acid sequence:
Subsequently, autologous pan-T cells were added to the co-culture to initiate TCR stimulation.
In all cases, the TCRs were stimulated for 11 days, followed by overnight re-stimulation treatment with individual peptides. Cells were harvested and CD8+ T cells were isolated using negative selection magnetic bead capture techniques. These isolated CD8+ T cells were analyzed using the 10× genomics single cell sequencing platform.
Procedure
Day −2 (for Ad5 treatment) or Day −1 (for exogenous and endogenous treatment): Donor dendritic cells (DC) from the four healthy human donors described, above, were thawed in IMDM5, (IMDM5 Media: Gibco, 12440 supplemented with 5% human antibody serum Sigma, H3667) with 1× anti-aggregate (C.T.L., CTL-AA-001). The cells were washed by centrifuging at 1300 rpm for five minutes and resuspended in IMDM5 containing 50 ng/mL GM-CSF and IL-4. The cells were plated into 6 well plates at ˜1-1.5e6 cells/well in 2 mL IMDM5 and incubated overnight at 37° C./5% CO2.
Day −1 (for Ad5 treatment): DCs were removed from plates, and wells of same donors were combined. Cells were washed in IMDM5 and counted and resuspended at a density of 6e5 cells/mL. 50 μL of cells were seeded (for a density of 3e4 cells/well) to appropriate wells of clear, round bottom 96 well plates according to plate layout. At least one row/column was left empty around outside edges of plates and filled with 100 μL DPBS. In a BSL2+ environment, Ad5 constructs were diluted in IMDM5 media with 2× (final concentration: 5 μg/mL) Polybrene (Millipore, Cat #: TR-1003-G) to concentrations of 5,000 Multiplicity of Infection (MOI) per 50 μL. 50 μL of each construct was added to the appropriate wells of DCs according to plate layout. Plates were returned to incubator at 37° C./5% CO2 overnight.
Day 0 (for endogenous treatment): DCs were removed from the plates, and wells of same donors were combined. Cells were washed in IMDM5 and counted and resuspended at a density of 5e5 cells/mL. 500 μL of cells were seeded (for a density of 2.5e5 cells/well) to appropriate wells of 24 well plates according to plate layout. Following NanoJuice Kit Protocol, (Millipore Sigma, 71902), Opti-MEM was combined with core transfection reagent and transfection booster and incubated at room temperature (RT) for five minutes. DNA was added to the appropriate tubes and mixed by gentle pipetting. Tubes were incubated at RT for 15 minutes. 20 μL of each preparation was added dropwise to the appropriate wells as indicated in plate layout. Plates were rocked gently to mix and incubated at 37° C./5% CO2 for six hours. Wells were transferred from endogenous transfection pates to microcentrifuge tubes. Tubes were centrifuged at 1500×g for five minutes. Cells were resuspended in 500 μL DPBS. Washes were repeated for a total of two washes. After the final wash, cells were resuspended in 500 μL IMDM5. Cells were pipetted to mix and 65 μL was transferred to appropriate wells indicated in the plate layout. The corresponding donor PanT cells (i.e. the PanT cells from the same donor as the dendritic cells) were thawed in IMDM5 with 1× anti-aggregate and washed by centrifuging at 1300 rpm for five minutes. Cells were resuspended in IMDM5 at 3e6 cells/mL. 100 μL of prepared PanT donor cells was added to corresponding donor DC wells according to plate layout (for a density of 3e5 PanT cells/well. Final ratio 1:10 DC:PanT). 200 μL of DPBS was added to outer wells of the plate(s). Plates were returned to incubator at 37° C./5% CO2.
Day 0 (for exogenous treatment): DCs were removed from plates, and wells of same donors were combined. Cells were washed in IMDM5 and counted and resuspended at a density of 3e5 cells/mL. 100 μL of cells were seeded (for a density of 3e4 cells/well) to appropriate wells in clear 96 well round bottom plate(s) as indicated in the plate layout. Peptides were diluted to 20× final concentrations in IMDM5 and 5 μL of the diluted peptides were added to appropriate exogenous wells according to the plate layout. 200 μL DPBS was added to outside wells and incubated at 37° C./5% CO2 for one and a half to two hours. After incubation, plates were placed in a room temperature irradiator for a calculated time to achieve the desired dose of 50 grays of radiation. Corresponding donor PanT cells were thawed in IMDM5 with 1× anti-aggregate and washed by centrifuging at 1300 rpm for five minutes. Cells were resuspended in IMDM5 at 3e6 cells/mL. 100 μL of prepared PanT donor cells were added to corresponding donor DC wells according to plate layout (for a density of 3e5 PanT cells/well. Final ratio 1:10 DC:PanT). The plates were returned to the incubator at 37° C./5% CO2.
Day 0 (for Ad5 treatment): After the plates were incubated with Ad5 for ˜20 hours they were removed from the incubator. The plates were washed by centrifuging at 300×g for five minutes and pellets were resuspended in DPBS (Gibco, 14140). Washes were repeated for a total of three washes. After the final wash, cells were resuspended in 100 μL IMDM5. Corresponding donor PanT cells were thawed in IMDM5 with 1× anti-aggregate. Cells were washed by centrifuging at 1300 rpm for five minutes and then were resuspended in IMDM5 at 3e6 cells/mL. 100 μL of prepared PanT donor cells were added to corresponding donor DC wells according to the plate layout (for a density of 3e5 PanT cells/well. Final ratio 1:10 DC:PanT). 200 μL of DPBS was added to outer wells of plates. Plates were returned to the incubator at 37° C./5% CO2.
Day 3 and following Day 3 (for all treatments): IMDM5 media was prepared with IL-2, IL-7, and IL-15 at 2× final concentrations. (Final concentrations: IL-2: 50 IU/mL, IL-7: 25 ng/mL, and IL-15: 25 ng/mL). 100 μL media was removed from each well and replaced with prepared IMDM5 cytokine media. Plates were gently rocked to mix and returned to the incubator at 37° C./5% CO2. This protocol was repeated every 2-3 days after, up to day 12, with the exception that IL-2 concentration was increased to 100 IU/mL.
Day 11: (If restim fell on a day when media needs to be refreshed, refresh was performed first). Peptides were prepared corresponding to exogenous, endogenous, or Ad5 constructs in IMDM5 for the restim of the PanT cells. For single peptide restim a final concentration of 5 ug/mL was used. For the CALR peptide pool, each peptide was used at a final concentration of 1 ug/mL (peptides were pooled at 500× final concentration prior to adding to plates). DMSO control was prepared in IMDM5 to match the highest concentration of peptide used (5 μg/mL). 5 μL of each preparation were added to appropriate wells. The plates were gently rocked to mix. The plates were returned to the incubator and incubated overnight (˜18 hours) at 37° C./5% CO2.
The sequences for the CALR peptide pool are provided in Table 1:
Day 12 (for all treatments): A vial of PanT cells were thawed from each donor (to be used as baselines) in IMDM5 with 1× anti-aggregate and washed by centrifuging at 1300 rpm for five minutes and resuspend in IMDM5. Replicates of each sample were combined into tubes on ice and centrifuged at 1500 rpm for five minutes at 4° C. Supernatant was removed and 300 μL running buffer was added to wash. The tubes were centrifuged again, and supernatant was removed. The antibody cocktail provided in the CD8 T cells negative selection bead kit was prepared, scaled accordingly (40 μL buffer and 10 μL antibody per sample). The antibody cocktail included: biotin-conjugated monoclonal anti-human antibodies against CD4, CD15, CD16, CD19, CD34, CD36, CD56, CD123, TCR γ/δ, and CD235a (Glycophorin A), MicroBeads conjugated to monoclonal anti-CD14 antibody (isotype: mouse IgG2a), anti-CD61 antibody (isotype: mouse IgG1), and anti-biotin antibody (isotype: mouse IgG1).
50 μL prepared antibody cocktail was added to each sample and incubated for five minutes on ice. Bead cocktail was prepared and scaled accordingly (30 μL buffer and 20 μL beads per sample). 50 μL was added to each sample and incubate for ten minutes on ice. 400 μL buffer was added to bring sample volumes to 500 μL. 400 μL buffer was added to baseline samples to bring volumes to 500 μL. The Manual CD8+ Separation protocol (MACS Miltenyi Biotec, 130-096-495) was followed. Columns and tubes (15 mL) were set up and rinsed with 3 mL buffer to pre-wet. 500 μL of cells was added and flow through was collected. Cells were washed with 3 mL buffer and collected. Cells were counted and washed by centrifuging at 1500 rpm for five minutes. The cells were resuspended in PBS at 1e4 cells/mL.
Single Cell V(D)J 10× Genomics Protocols
The Single Cell V(D)J (variable, diverse, and joining) 10× genomics protocols (Chromium™ Single Cell V(D)J Reagent Kits User Guide—CG000086) was used for measuring immune repertoire information and gene expression from the same cell. Cells obtained from the 12 Day Assay were washed and resuspended in 1× phosphate-buffered saline (PBS) containing 0.04% bovine serum albumin (BSA) to achieve target cell concentration (˜20,000 cells/sample). Gel Bead-in-Emulsions (GEMs) were generated by combining barcoded single cell VDJ 5′ Gel beads, a Master Mix with cells, and partitioning oil on a 10× Chromium Controller Single Cell A Chip. Following GEM generation, the Gel Bead dissolved, and any co-partitioned cell was lysed. Oligonucleotides containing sequencing primers, 10× cell barcode, 10× unique molecular identifier (UMI) barcode, and template switch oligo (TSO) were released and mixed with the cell lysate and Master Mix containing reverse transcription (RT) reagents. Incubation of GEM-RT produced barcoded full-length cDNA from mRNA. 10× barcoded full-length cDNA from GEM-RT was purified and amplified via polymerase chain reaction (PCR) (14 cycles) with primers against the 5′ and 3′ ends added during GEM-RT. Full Length VDJ segments were enriched from amplified cDNA with primers specific to TCR region. Enzymatic fragmentation and size selection were used to optimize the cDNA amplicon size prior to 5′ gene expression (GEX) library construction. P5 (AATGATACGGCGACCACCGA) (SEQ ID NO: 703), P7 (CAAGCAGAAGACGGCATACGAGAT) (SEQ ID NO: 704), a sample index) (SEQ ID NO: 705), and Illumina R2 (AGATCGGAAGAGCACACGTCTGAACTCCAGTCAC) (SEQ ID NO: 706) sequence were added via End Repair, A tailing, Adaptor Ligation and Sample Index PCR. The final libraries contained the P5 and P7 priming sites used in Illumina Sequencing. Pooled/normalized libraries were sequenced at the recommended depth/run parameters. Libraries sequenced on NovaSeq produced high quality data with expected Q30 (sequencing Analysis Viewer) scores. Multiplexed sequences were de-multiplexed and analyzed using The Chromium Single Cell Immune Profiling Software Suite.
Identifying Expanded TCRs
To identify TCRs enriched due to antigen stimulation, T-cell receptor alpha/beta (TCR α/β) sequences from T-cell populations treated with either endogenous minigene or by Ad5 transduction were compared against corresponding sequences from T-cell populations obtained by stimulation against an empty minigene (i.e. no antigen) or an Ad5 null construct (i.e. Ad5 transfection without an antigen cassette). For TCRα/β sequences obtained from stimulation by exogenously loaded dendritic cells (DC's), the corresponding DMSO treated controls (i.e. DMSO+no antigen stimulation) were used. TCR α/β sequences present in cell populations enriched at a false discovery rate (FDR) of <0.0003 and a minimum of 2 fold increase in TCR proportions compared to controls were selected for further analysis. TCR α/β sequences that were highly abundant (˜1.0% or higher) both in the controls and treatment conditions and that were significantly enriched were also chosen for further examination.
Identifying Expanded TCR Sequences from Activated T Cells
To further triage the number of TCRs for experimental validation, single cell RNA (scRNA) sequencing data was obtained from the same cells. The T-cell activation markers considered for mRNA based gating of T cells were TNF, GZMB, CD69 and PRF1 (Perforin). These markers and their cutoffs are listed in Table 2, below. TCRs which were both clonally expanded and present in cells with higher than the cutoff mRNA expression in all 4 genes were given the highest priority. In addition, TCRs that shared sequence identity with known self-antigens or viral antigens were eliminated from further consideration. The IEDB (www.iedb.org) public repository database was used to identify the self/viral antigen responsive TCR sequences.
scRNA Analysis
The Seurat R package was used to perform the scRNA sequencing analysis. The samples were first quality controlled using high mitochondrial content (>5%), excessively low (<300) counts, and library size criteria. Cells that failed to pass quality control (QC) thresholds were removed. After QC filtering, the SCTransform algorithm was used to normalize and transform raw RNA read counts. Samples pre- and post-12 day expansion were combined using the sample Integration method implemented in Seurat. A principal component analysis (PCA), followed by uniform manifold approximation and projection (UMAP) clustering on the first 30 principal components was performed. The normalized mRNA data was then utilized, and the expression profiles of various T cell activation markers were examined to establish the cutoff criteria. The activation profiles showed a generally bimodal distribution, and the cutoff was chosen at the point of the trough between the two neighboring peaks of the bimodal distribution. See table 1 for cut-off values.
Table 3 provides donor number, treatment condition, the V(D)J genes, and the constant genes for thirty-four sequenced T-cell receptors.
Table 4 provides the full length alpha chain sequences as well as the nucleotide (NT) and amino acid (AA) sequences of the alpha chain CDR1, CDR2 and CDR3, defined by IMGT, for each of the thirty-four T-cell receptors described in Table 3. Table 5 provides the full length beta chain sequences as well as the nucleotide and amino acid sequences of the beta chain CDR1, CDR2 and CDR3, defined by IMGT, for each of the thirty-four T-cell receptors described in Table 3. For both Table 4 and Table 5, the TCR alpha or beta (full length) amino acid sequence that was derived from the scRNA analysis had an incomplete constant gene sequence. In order to complete this, alignments of the incomplete constant region sequence were performed against the canonical TCRB1 and TCRB2 sequences, and the additional missing portion appended. In two instances, the alignments had incomplete query coverage, in these cases the overhanging portion in the incomplete gene was deleted and the correct TCRB sequence appended. For the TCR alpha gene, the missing portion of the TCR alpha gene sequence was appended. The full constant gene is identified by name in Table 3. The VJ/VDJ portion of the TCR alpha or beta (full length) amino acid sequence is underlined, the constant region is in underlined and bold, and the unmarked sequence is the leader sequence. For the corresponding nucleotide sequences in Tables 4 and 5, the leader sequence is underlined, the constant region is underlined and bolded, and the VJ/VDJ portion of the full length TCR nucleotide sequence is unmarked.
VEQNPLFLSMQ
EGKNYTIYCNY
STTSDRLYWYR
QDPGKSLESLF
VLLSNGAVKQE
GRLMASLDTKA
GCAATGATTCTGTGGCTTCA
RLSTLHITAAV
ACTAGACCGGTTAAGTGGAG
HDLSATYFCAA
RSYNTDKLIFGT
GTRLQVFPNIQ
VAQPEDQVNV
AEGNPLTVKCT
YSVSGNPYLFW
YVQYPNRGLQF
LLKYITGDNLV
KGSYGFEAEFN
KSQTSFHLKKP
SALVSDSALYF
CAVRDMGGSA
RQLTFGSGTQL
TCTCGATGCTTGCGATGCTCT
TCACATTGAGTGGGCTGAGA
QPISMDSYEGQ
EVNITCSHNNIA
TNDYITWYQQF
PSQGPRFIIQGY
TGGCGAGAGTGATCGTGTTC
KTKVTNEVASL
CTGACCCTGAGTACTTTGAG
FIPADRKSSTLS
CCTTGCTAAGACCACCCAGC
LPRVSLSDTAV
YYCLVQTFSGS
ARQLTFGSGTQ
NEVEQSPQNLT
AQEGEFITINCS
YSVGISALHWL
QQHPGGGIVSL
FMLSSGKKKHG
RLIATINIQEKH
SSLHITASHPRD
SAVYICAVSTS
MYSGGGADGL
TFGKGTHLIIQP
GAAGATCCGGCAATTTTTGT
TGGCTATTTTGTGGCTTCAGC
LSVPEGAIASLN
CTYSDRGSQSF
FWYRQYSGKSP
ELIMSIYSNGDK
EDGRFTAQLNK
AAATCCTTGAGAGTTTTACT
ASQYVSLLIRDS
AGTGATCCTGTGGCTTCAGT
QPSDSATYLCA
TGAGCTGGGTTTGGAGCCAA
VKGGYALNFG
KGTSLLVTPHI
TVTQSQPEMSV
QEAETVTLSCT
YDTSESDYYLF
WYKQPPSRQMI
CCTGTGGGCACTTGTGATCT
LVIRQEAYKQQ
CCACCTGTCTTGAATTTAGC
NATENRFSVNF
ATGGCTCAGACAGTCACTCA
QKAAKSFSLKIS
DSQLGDAAMY
FCAYSPPLPGN
QFYFGTGTSLT
VEQNPLFLSMQ
EGKNYTIYCNY
STTSDRLYWYR
QDPGKSLESLF
GAAGCTACTAGCAATGATTC
VLLSNGAVKQE
TGTGGCTTCAACTAGACCGG
GRLMASLDTKA
TTAAGTGGAGAGCTGAAAGT
RLSTLHITAAV
HDLSATYFCAG
EPDSNYQLIWG
AGTKLIIKPDIQ
QQKEVEQNSGP
LSVPEGAIASLN
CTYSDRGSQSF
FWYRQYSGKSP
ELIMSIYSNGDK
EDGRFTAQLNK
ASQYVSLLIRDS
QPSDSATYLCA
VGGAGGYQKV
TCCTTGAGAGTTTTACTAGTG
TFGTGTKLQVIP
ATCCTGTGGCTTCAGTTGAG
CTGGGTTTGGAGCCAACAGA
(SEQ
LEQPSEVTAVE
GAIVQINCTYQ
TSGFYGLSWYQ
CATGGTATCCTGCAGCAGAT
QHDGGAPTFLS
GTGGGGAGCTTTCCTTCTCTA
YNALDGLEETG
TGTTTCCATGAAGATGGGAG
RFSSFLSRSDSY
GCACTGCAGGACAAAGCCTT
GYLLLQELQMK
DSASYFCAVRD
PLSGGYNKLIFG
AGTRLAVHPYI
(SEQ ID NO: 57)
PNSMESKEEEP
VHLPCNHSTISG
TDYIHWYRQLP
SQGPEYVIHGL
TSNVNNRMASL
TGGTGACAAGCATTACTGTA
AIAEDRKSSTLI
CTCCTATCTTTGGGTATTATG
LHRATLRDAAV
GGTGATGCTAAGACCACACA
YYCILYSGGGA
DGLTFGKGTHL
TEMTATEGAIV
QINCTYQTSGF
NGLFWYQQHA
GEAPTFLSYNV
LDGLEEKGRFS
SFLSRSKGYSYL
LLKELQMKDSA
SYLCAGLFGNT
PLVFGKGTRLS
GGAATGCAGTGCCAGGCTCG
TGGTATCCTGCAGCAGATGT
GGGGAGTTTTCCTTCTTTATG
TTTCCATGAAGATGGGAGGC
ACTACAGGACAAAACATTGA
TVTQSQPEMSV
QEAETVTLSCT
YDTSESDYYLF
GGCATGCCCTGGCTTCCTGT
WYKQPPSRQMI
GGGCACTTGTGATCTCCACC
LVIRQEAYKQQ
TGTCTTGAATTTAGCATGGCT
NATENRFSVNF
QKAAKSFSLKIS
DSQLGDAAMY
FCAYFNTDKLIF
GTGTRLQVFPN
(SEQ
LSVPEGAIVSLN
CTYSNSAFQYF
MWYRQYSRKG
PELLMYTYSSG
NKEDGRFTAQV
DKSSKYISLFIR
DSQPSDSATYL
CAMGGQGGAD
TTGAGAGTTTTACTGGTGAT
GLTFGKGTHLII
CCTGTGGCTTCAGTTAAGCT
GGGTTTGGAGCCAACAGAAG
(SEQ ID
LNQSPQSMFIQE
GEDVSMNCTSS
SIFNTWLWYKQ
TTATTAATAATCTTGTGGATG
DPGEGPVLLIAL
CAGCTGACATGGGTCAGTGG
YKAGELTSNGR
LTAQFGITRKDS
FLNISASIPSDV
GIYFCAGPKRE
YGNKLVFGAGT
PSTVASSEGAV
VEIFCNHSVSN
AYNFFWYLHFP
GCAPRLLVKGS
AGCACTCTGGGGGCGGTGTG
KPSQQGRYNM
GCTAGGGCTTCTCCTCAACT
TYERFSSSLLIL
CTCTCTGGAAGGTTGCAGAA
QVREADAAVY
AGCAAGGACCAAGTGTTTCA
YCAVEDNNNN
DMRFGAGTRLT
(SEQ ID NO: 99)
LEQPSEVTAVE
GAIVQINCTYQ
TSGFYGLSWYQ
QHDGGAPTFLS
YNALDGLEETG
RFSSFLSRSDSY
GYLLLQELQMK
DSASYFCAVNN
NARLMFGDGT
GGAGCTTTCCTTCTCTATGTT
TCCATGAAGATGGGAGGCAC
TGCAGGACAAAGCCTTGAGC
(SEQ ID NO:
SFLSRSKGYSYL
LLKELQMKDSA
SYLCAVRDPGN
DMRFGAGTRLT
GGCTCAGGAACTGGGAATGC
AGTGCCAGGCTCGTGGTATC
CTGCAGCAGATGTGGGGAGT
TTTCCTTCTTTATGTTTCCAT
GAAGATGGGAGGCACTACA
(SEQ ID NO:
GTTAGCTTGCTGTGGGCAGT
CGTGGTCTCCACCTGTCTTGA
ATCCGGCATGGCCCAGACAG
MYFCAFGGETS
GSRLTFGEGTQ
AGGCAAGTGGCGAGAGTGAT
CGTGTTCCTGACCCTGAGTA
IKVTNEVASLFI
CTTTGAGCCTTGCTAAGACC
PADRKSSTLSLP
RVSLSDTAVYY
CLVGDKGSNY
QLIWGAGTKLII
EQHPSTLSVQE
GDSAVIKCTYS
DSASNYFPWYK
QELGKRPQLIID
IRSNVGEKKDQ
RIAVTLNKTAK
HFSLHITETQPE
DSAVYFCAASY
LPDMRFGAGTR
ATCCATTCGAGCTGTATTTAT
ATTCCTGTGGCTGCAGCTGG
ACTTGGTGAATGGAGAGAAT
NNSI
IPEDTFFPSPES
SCDVKLVEKSF
ETDTNLNFQN
LSVIGFRILLL
KVAGFNLLMT
LRLWSS
AQKVTQAQTEI
SVVEKEDVTLD
CVYETRDTTYY
LFWYKQPPSGE
LVFLIRRNSFDE
QNEISGRYSWN
FQKSTSSFNFTI
TASQVVDSAVY
AACATGCTGACTGCCAGCCT
FCALSELKAAG
GTTGAGGGCAGTCATAGCCT
NKLTFGGGTRV
CCATCTGTGTTGTATCCAGC
ATGGCTCAGAAGGTAACTCA
NNSII
PEDTFFPSPESS
CDVKLVEKSF
ETDTNLNFQN
LSVIGFRILLL
KVAGFNLLMT
LRLWSS
(SEQ
PISMDSYEGQE
VNITCSHNNIAT
NDYITWYQQFP
SQGPRFIIQGYK
TKVTNEVASLFI
AGTGATCGTGTTCCTGACCC
PADRKSSTLSLP
TGAGTACTTTGAGCCTTGCT
RVSLSDTAVYY
CLVGDKVYGG
SQGNLIFGKGT
NN
SIIPEDTFFPSP
ESSCDVKLVEK
SFETDTNLNFQ
NLSVIGFRILLL
KVAGFNLLMT
LRLWSS
KITQTQPGMFV
QEKEAVTLDCT
YDTSDPSYGLF
WYKQPSSGEMI
FLIYQGSYDQQ
NATEGRYSLNF
QKARKSANLVI
SASQLGDSAMY
FCAMREGQDSS
YKLIFGSGTRLL
GAAGGTGGTCACAGCTTCAC
TGTGGCTAGGACCTGGCATT
NNSII
PEDTFFPSPESS
CDVKLVEKSF
ETDTNLNFQN
LSVIGFR1LLL
KVAGFNLLMT
LRLWSS
(SEQ ID
CATCAGTGCTGATTCTGTGG
CTTCAGCCAGACTGGGTAAA
CAGTCAACAGAAGAATGATG
SLHIVPSQPGDS
AVYFCAARIYG
GSQGNLIFGKG
TKLSVKPNIQN
NN
SIIPEDTFFPSP
ESSCDVKLVEK
SFETDTNLNFQ
NLSVIGFRILLL
KVAGFNLLMT
LRLWSS
(SEQ ID NO: 162)
KITQTQPGMFV
QEKEAVTLDCT
YDTSDPSYGLF
WYKQPSSGEMI
FLIYQGSYDQQ
NATEGRYSLNF
QKARKSANLVI
SASQLGDSAMY
FCAMREGEPSG
ATGTCACTTTCTAGCCTGCTG
SARQLTFGSGT
AAGGTGGTCACAGCTTCACT
GTGGCTAGGACCTGGCATTG
NN
SIIPEDTFFPSP
ESSCDVKLVEK
SFETDTNLNFQ
NLSVIGFRILLL
KVAGFNLLMT
LRLWSS
QNSPSLSVQEG
RISILNCDYTNS
MFDYFLWYKK
ATGGCCATGCTCCTGGGGGC
YPAEGPTFLISIS
ATCAGTGCTGATTCTGTGGC
SIKDKNEDGRF
TVFLKKSAKHL
AGTCAACAGAAGAATGATGA
SLHIVPSQPGDS
AVYFCAVNAG
GTSYGKLTFGQ
NN
SIIPEDTFFPSP
ESSCDVKLVEK
SFETDTNLNFQ
NLSVIGFRILLL
KVAGFNLLMT
LRLWSS
TQLGSHVSVSE
GALVLLRCNYS
SSVPPYLFWYV
QYPNOGLQLLL
KYTTGATLVKG
INGFEAEFKKSE
TSFHLTKPSAH
MSDAAEYFCA
VSEKFSGGYNK
LIFGAGTRLAV
GCTGCTCGTCCCAGTGCTCG
NNSIIPE
AGGTGATTTTTACCCTGGGA
DTFFPSPESSC
GGAACCAGAGCCCAGTCGGT
DVKLVEKSFET
DTNLNFQNLSV
IGFRILLLKVA
GFNLLMTLRL
WSS
AQKVTQAQTEI
SVVEKEDVTLD
CVYETRDTTYY
LFWYKQPPSGE
LVFLIRRNSFDE
ATGAACATGCTGACTGCCAG
QNEISGRYSWN
CCTGTTGAGGGCAGTCATAG
FQKSTSSFNFTI
CCTCCATCTGTGTTGTATCCA
TASQVVDSAVY
GCATGGCTCAGAAGGTAACT
FCALSEAKDDK
IIFGKGTRLHILP
NNSIIPEDT
FFPSPESSCDV
KLVEKSFETDT
NLNFQNLSVIG
FRILLLKVAGF
NLLMTLRLWS
S
QNSPSLSVQHQ
RISILNCDYTNS
MFDYFLWYKK
YPAEGPTFLISIS
SIKDKNEDGRF
TVFLNKSAKHL
SLHIVPSQPGDS
AVYFCAIYNQG
GKLIFGQGTELS
NNSII
PEDTFFPSPESS
CDVKLVEKSF
ETDTNLNFQN
LSVIGFRILLL
KVAGFNLLMT
LRLWSS
VTQMEGPVTLS
EEAFLTINCTYT
TGAACTATTCTCCAGGCTTA
ATGYPSLFWYV
GTATCTCTGATACTCTTACTG
QYPGEGLQLLL
CTTGGAAGAACCCGTGGAAA
KATKADDKGS
NKGFEATYRKE
TTSFHLEKGSV
QVSDSAVYFCA
LMTDYKLSFGA
NNSIIPEDTFF
PSPESSCDVKL
VEKSFETDTNL
NFQNLSVIGFR
ILLLKVAGFNL
LMTLRLWSS
LSVPEGAIASLN
CTYSDRGSQSF
FVVYRQYSGKSP
ELIMSIYSNGDK
EDGRFTAQLNK
ATCCTTGAGAGTTTTACTAGT
ASQYVSLLIRDS
GATCCTGTGGCTTCAGTTGA
QPSDSATYLCA
GCTGGGTTTGGAGCCAACAG
VRSVGVSGGYN
KLIFGAGTRLA
NNSII
PEDTFFPSPESS
CDVKLVEKSF
ETDTNLNFQN
LSVIGFRILLL
KVAGFNLLMT
LRLWSS
PISMDSYEGQE
VNITCSHNNIAT
NDYITWYQQFP
SQGPRFIIQGYK
GAGTGATCGTGTTCCTGACC
TKVTNEVASLFI
CTGAGTACTTTGAGCCTTGCT
PADRKSSTLSLP
RVSLSDTAVYY
CLVLLSSNTGK
LIFGQGTTLQV
NNSIIPE
DTFFPSPESSC
DVKLVEKSFET
DTNLNFQNLSV
IGFRILLLKVA
GFNLLMTLRL
WSS
TVTQSQPEMSV
QEAETVTLSCT
YDTSESDYYLF
GGCATGCCCTGGCTTCCTGT
WYKQPPSRQMI
GGGCACTTGTGATCTCCACC
LVIRQEAYKQQ
TGTCTTGAATTTAGCATGGCT
NATENRFSVNF
QKAAKSFSLKIS
DSQLGDAAMY
FCAYRSANNND
MRFGAGTRLTV
NNSIIPE
DTFFPSPESSC
DVKLVEKSFET
DTNLNFQNLSV
IGFRILLLKVA
GFNLLMTLRL
WSS
RKEVEQDPGPF
NVPEGATVAFN
CTYSNSASQSFF
WYRQDCRKEP
KLLMSVYSSGN
EDGRFTAQLNR
ASQYISLLIRDS
KLSDSATYLCV
VFPGGSYIPTFG
ATGATATCCTTGAGAGTTTT
ACTGGTGATCCTGTGGCTTC
AGTTAAGCTGGGTTTGGAGC
CAACGGAAGGAGGTGGAGC
NNSIIPEDTFF
PSPESSCDVKL
VEKSFETDTNL
NFQNLSVIGFR
ILLLKVAGFNL
LMTLRLWSS
VTQTPRHKVTE
MGQEVTMRCQ
PILGHNTVFWY
RQTMMQGLEL
LAYFRNRAPLD
DSGMPKDRFSA
EMPDATLATLK
IQPSEPRDSAVY
FCASGLALTEG
GWYEQYFGPG
TRLTVTEDLKN
VFPPEVAVFEP
SEAEISHTQKA
TLVCLATGFYP
CACCAGGCTCCTCTGCTGTG
DHVELSWWVN
TGGTTCTTTGTCTCCTGGGAG
GKEVHSGVST
AAGAGCTTATAGATGCTAGA
DPCIPLKEQPA
LNDSRYCLSSR
LRVSATFWQN
PRNHFRCQVQ
FYGLSENDEW
TQDRAKPVTQI
VSAEAWGRAD
CGFTSESYQQ
GVLSATILYEI
LLGKATLYAV
LVSALVLMAM
VKRKDSRG
GTTCCCACCCGAGGTCGCT
GTGTTTGAGCCATCAGAAG
CAGAGATCTCCCACACCCA
AAAGGCCACACTGGTATGC
CTGGCCACAGGCTTCTACC
CCGACCACGTGGAGCTGAG
CTGGTGGGTGAATGGGAA
GGAGGTGCACAGTGGGGT
CAGCACAGACCCGCAGCCC
CTCAAGGAGCAGCCCGCCC
TCAATGACTCCAGATACTG
CCTGAGCAGCCGCCTGAGG
GTCTCGGCCACCTTCTGGC
AGAACCCCCGCAACCACTT
CCGCTGTCAAGTCCAGTTC
TACGGGCTCTCGGAGAATG
ACGAGTGGACCCAGGATAG
GGCCAAACCTGTCACCCAG
ATCGTCAGCGCCGAGGCCT
GGGGTAGAGCAGACTGTG
GCTTCACCTCCGAGTCTTA
CCAGCAAGGGGTCCTGTCT
GCCACCATCCTCTATGAGA
TCTTGCTAGGGAAGGCCAC
CTTGTATGCCGTGCTGGTC
AGTGCCCTCGTGCTGATGG
CCATGGTCAAGAGAAAGGA
TTCCAGAGGCTAG
(SEQ ID
VTQTPSHQVTQ
MGQEVILRCVPI
SNHLYFYWYR
QILGQKVEFLV
SFYNNEISEKSE
CTCGTATGCTGGGCAATTTTT
IFDDQFSVERPD
AGTCTCTTGAAAGCAGGACT
GSNFTLKIRSTK
CACAGAACCTGAAGTCACCC
LEDSAMYFCAS
LRWDGDNEQFF
GPGTRLTVLED
LKNVFPPEVAV
FEPSEAEISHT
QKATLVCLAT
GFYPDHVELS
WWVNGKEVH
SGVSTDPCIPLK
EQPALNDSRY
CLSSRLRVSAT
FWQNPRNHFR
CQVQFYGLSE
NDEWTQDRAK
PVT
Q
IVSAEAW
GRADCGFTSES
Y
QQ
GVLSATIL
YEILLGKATLY
TGTTCCCACCCGAGGTCGC
AVLVSALVLM
TGTGTTTGAGCCATCAGAA
AMVKRKDSRG
GCAGAGATCTCCCACACCC
AAAAGGCCACACTGGTATG
CCTGGCCACAGGCTTCTAC
CCCGACCACGTGGAGCTGA
GCTGGTGGGTGAATGGGA
AGGAGGTGCACAGTGGGG
TCAGCACAGACCCGCAGCC
CCTCAAGGAGCAGCCCGCC
CTCAATGACTCCAGATACT
GCCTGAGCAGCCGCCTGAG
GGTCTCGGCCACCTTCTGG
CAGAACCCCCGCAACCACT
TCCGCTGTCAAGTCCAGTT
CTACGGGCTCTCGGAGAAT
GACGAGTGGACCCAGGATA
GGGCCAAACCTGTCACCCA
GATCGTCAGCGCCGAGGCC
TGGGGTAGAGCAGACTGTG
GCTTCACCTCCGAGTCTTA
CCAGCAAGGGGTCCTGTCT
GCCACCATCCTCTATGAGA
TCTTGCTAGGGAAGGCCAC
CTTGTATGCCGTGCTGGTC
AGTGCCCTCGTGCTGATGG
CCATGGTCAAGAGAAAGGA
TTCCAGAGGCTAG
(SEQ ID
GAATGACCCCCTTGAGAGTA
CTGTTCCCCTATCACCGATGC
QTPKFQVLKTG
ACAGACCCAGAAGACCCCTC
QSMTLQCAQD
CATCCTGTAGCACCTGCCAT
MNHNSMYWYR
GAGCATCGGGCTCCTGTGCT
QDPGMGLRLIY
GTGTGGCCTTTTCTCTCCTGT
YSASEGTTDKG
GGGCAAGTCCAGTGAATGCT
EVPNGYNVSRL
NKREFSLRLES
AAPSQTSVYFC
ASRDRALNTEA
FFGQGTRLTVV
EDLNKVFPPEV
AVFEPSEAEIS
HTQKATLVCL
ATGFYPDHVE
LSWWVNGKE
VHSGVSTDPQP
LKEQPALNDS
RYCLSSRLRVS
ATFWQNPRNH
FRCQVQFYGL
SENDEWTQDR
AKPVTQIV
SAEAWGRADC
CTGAACAAGGTGTTCCCAC
GFTSVSYQQG
CCGAGGTCGCTGTGTTTGA
VLSATILYEILL
GCCATCAGAAGCAGAGATC
GKATLYAVLV
TCCCACACCCAAAAGGCCA
SALVLMAMVK
CACTGGTGTGCCTGGCCAC
RKDF
AGGCTTCTACCCCGACCAC
GTGGAGCTGAGCTGGTGG
GTGAATGGGAAGGAGGTG
CACAGTGGGGTCAGCACAG
ACCCGCAGCCCCTCAAGGA
GCAGCCCGCCCTCAATGAC
TCCAGATACTGCCTGAGCA
GCCGCCTGAGGGTCTCGGC
CACCTTCTGGCAGAACCCC
CGCAACCACTTCCGCTGTC
AAGTCCAGTTCTACGGGCT
CTCGGAGAATGACGAGTGG
ACCCAGGATAGGGCCAAAC
CCGTCACCCAGATCGTCAG
CGCCGAGGCCTGGGGTAG
AGCAGACTGTGGCTTTACC
TCGGTGTCCTACCAGCAAG
GGGTCCTGTCTGCCACCAT
CCTCTATGAGATCCTGCTA
GGGAAGGCCACCCTGTATG
CTGTGCTGGTCAGCGCCCT
TGTGTTGATGGCCATGGTC
AAGAGAAAGGATTTCTGA
LKTGQSMTLQC
AQDMNHEYMS
WYRQDPGMGL
GCCTCTCCCTCCCTGGGGCC
RLIHYSVGAGIT
CAGGCAGGGAGAATGTCTCA
DQGEVPNGYN
GAATGACTTCCTTGAGAGTC
VSRSTTEDFPLR
CTGCTCCCCTTTCATCAATGC
LLSAAPSQTSV
ACAGATACAGAAGACCCCTC
YFCASRNGGTL
CGTCATGCAGCATCTGCCAT
IYEQYFGPGTRL
GAGCATCGGCCTCCTGTGCT
TVTEDLKNVFP
GTGCAGCCTTGTCTCTCCTGT
PEVAVFEPSEA
GGGCAGGTCCAGTGAATGCT
EISHT
Q
KATLV
CLATGFYPDH
VELSWWVNG
KEVHSGVSTDP
QPLKEQPALN
DSRYCLSSRLR
VSATFWQNPR
NHFRCQVQFY
GLSENDEWTQ
DRAKPVTQIVS
AEAWGRADCG
FTSESYQQGVL
SATILYEILLG
KATLYAVLVS
ALVLMAMVK
RKDSRG
GACCTGAAAAACGTGTTCC
CACCCGAGGTCGCTGTGTT
TGAGCCATCAGAAGCAGAG
ATCTCCCACACCCAAAAGG
CCACACTGGTATGCCTGGC
CACAGGCTTCTACCCCGAC
CACGTGGAGCTGAGCTGGT
GGGTGAATGGGAAGGAGG
TGCACAGTGGGGTCAGCAC
AGACCCGCAGCCCCTCAAG
GAGCAGCCCGCCCTCAATG
ACTCCAGATACTGCCTGAG
CAGCCGCCTGAGGGTCTCG
GCCACCTTCTGGCAGAACC
CCCGCAACCACTTCCGCTG
TCAAGTCCAGTTCTACGGG
CTCTCGGAGAATGACGAGT
GGACCCAGGATAGGGCCA
AACCTGTCACCCAGATCGT
CAGCGCCGAGGCCTGGGG
TAGAGCAGACTGTGGCTTC
ACCTCCGAGTCTTACCAGC
AAGGGGTCCTGTCTGCCAC
CATCCTCTATGAGATCTTG
CTAGGGAAGGCCACCTTGT
ATGCCGTGCTGGTCAGTGC
CCTCGTGCTGATGGCCATG
GTCAAGAGAAAGGATTCCA
GAGGCTAG
GVTQTPRYLIK
TRGQQVTLSCS
PISGHRSVSWY
QQTPGQGLQFL
FEYFSETQRNK
GNFPGRFSGRQ
FSNSRSEMNVS
TLELGDSALYL
CASRQRTELEA
FFGQGTRLTVV
EDLNKVFPPEV
AVFEPSEAEIS
HTQKATLVCL
ATGFYPDHVE
LSWWVNGKE
GCTCCAGGCTGCTCTGTTGG
VHSGVSTDPQP
GTGCTGCTTTGTCTCCTGGGA
LKECIPALNDS
GCAGGCCCAGTAAAGGCTGG
RYCLSSRLRVS
ATFWQNPRNH
FRCQVQFYGL
SENDEWTQDR
AKPVTQIVSAE
AWGRADCGFT
SVSYQQGVLSA
TILYEILLGKA
TLYAVLVSALV
LMAMVKRKD
F
CAAGGTGTTCCCACCCGAG
GTCGCTGTGTTTGAGCCAT
CAGAAGCAGAGATCTCCCA
CACCCAAAAGGCCACACTG
GTGTGCCTGGCCACAGGCT
TCTACCCCGACCACGTGGA
GCTGAGCTGGTGGGTGAAT
GGGAAGGAGGTGCACAGT
GGGGTCAGCACAGACCCG
CAGCCCCTCAAGGAGCAGC
CCGCCCTCAATGACTCCAG
ATACTGCCTGAGCAGCCGC
CTGAGGGTCTCGGCCACCT
TCTGGCAGAACCCCCGCAA
CCACTTCCGCTGTCAAGTC
CAGTTCTACGGGCTCTCGG
AGAATGACGAGTGGACCCA
GGATAGGGCCAAACCCGTC
ACCCAGATCGTCAGCGCCG
AGGCCTGGGGTAGAGCAG
ACTGTGGCTTTACCTCGGT
GTCCTACCAGCAAGGGGTC
CTGTCTGCCACCATCCTCT
ATGAGATCCTGCTAGGGAA
GGCCACCCTGTATGCTGTG
CTGGTCAGCGCCCTTGTGT
TGATGGCCATGGTCAAGAG
AAAGGATTTCTGA
VTQTPSHQVTQ
MGQEVILRCVPI
SNHLYFYWYR
QILGQKVEFLV
SFYNNEISEKSE
TCGTATGCTGGGCAATTTTTA
IFDDQFSVERPD
GTCTCTTGAAAGCAGGACTC
GSNFTLKIRSTK
ACAGAACCTGAAGTCACCCA
LEDSAMYFCAS
SEAPDRGTYEQ
YFGPGTRLTVT
EDLKNVFPPEV
AVFEPSEAEIS
HT
Q
KATLVCL
ATGFYPDHVE
LSWWVNGKE
VHSGVSTDPQP
LKEQPALNDS
RYCLSSRLRVS
ATFWQNPRNH
FRCQVQFYGL
SENDEWTQDR
AKPVTQIVSAE
AWGRADCGFT
SESYQQGVLSA
TILYEILLGKA
AAAACGTGTTCCCACCCGA
TLYAVLVSALV
GGTCGCTGTGTTTGAGCCA
LMAMVKRKDS
TCAGAAGCAGAGATCTCCC
RG
ACACCCAAAAGGCCACACT
GGTGTGCCTGGCCACAGGC
TTCTACCCCGACCACGTGG
AGCTGAGCTGGTGGGTGAA
TGGGAAGGAGGTGCACAG
TGGGGTCAGCACAGACCCG
CAGCCCCTCAAGGAGCAGC
CCGCCCTCAATGACTCCAG
ATACTGCCTGAGCAGCCGC
CTGAGGGTCTCGGCCACCT
TCTGGCAGAACCCCCGCAA
CCACTTCCGCTGTCAAGTC
CAGTTCTACGGGCTCTCGG
AGAATGACGAGTGGACCCA
GGATAGGGCCAAACCTGTC
ACCCAGATCGTCAGCGCCG
AGGCCTGGGGTAGAGCAG
ACTGTGGCTTCACCTCCGA
GTCTTACCAGCAAGGGGTC
CTGTCTGCCACCATCCTCT
ATGAGATCTTGCTAGGGAA
GGCCACCTTGTATGCCGTG
CTGGTCAGTGCCCTCGTGC
TGATGGCCATGGTCAAGAG
AAAGGATTCCAGAGGCTAG
GITQSPRYKITE
TGRQVTLMCH
CAGGCTCTTCTTCTATGTGGC
QTWSHSYMFW
CCTTTGTCTGCTGTGGGCAG
YRQDLGHGLRL
GACACAGGGATGCTGGAATC
IYYSAAADITD
KGEVPDGYVVS
RSKTENFPLTLE
SATRSQTSVYF
CASSELGRGFY
EQYFGPGTRLT
VTEDLKNVFPP
EVAVFEPSEAE
ISHTQKATLVC
LATGFYPDHV
ELSWWVNGKE
VHSGVSTDPQP
LKEQPALNDS
RYCLSSRLRVS
ATFW
Q
NPRNH
FRC
Q
V
Q
FYGL
SENDEWTQDR
AKPVTQIVSAE
AAAAACGTGTTCCCACCCG
AWGRADCGFT
AGGTCGCTGTGTTTGAGCC
SESYQQGVLSA
ATCAGAAGCAGAGATCTCC
TILYEILLGKA
CACACCCAAAAGGCCACAC
TLYAVLVSALV
TGGTGTGCCTGGCCACAGG
LMAMVKRKDS
CTTCTACCCCGACCACGTG
RG
GAGCTGAGCTGGTGGGTG
AATGGGAAGGAGGTGCAC
AGTGGGGTCAGCACAGACC
CGCAGCCCCTCAAGGAGCA
GCCCGCCCTCAATGACTCC
AGATACTGCCTGAGCAGCC
GCCTGAGGGTCTCGGCCAC
CTTCTGGCAGAACCCCCGC
AACCACTTCCGCTGTCAAG
TCCAGTTCTACGGGCTCTC
GGAGAATGACGAGTGGAC
CCAGGATAGGGCCAAACCT
GTCACCCAGATCGTCAGCG
CCGAGGCCTGGGGTAGAG
CAGACTGTGGCTTCACCTC
CGAGTCTTACCAGCAAGGG
GTCCTGTCTGCCACCATCC
TCTATGAGATCTTGCTAGG
GAAGGCCACCTTGTATGCC
GTGCTGGTCAGTGCCCTCG
TGCTGATGGCCATGGTCAA
GAGAAAGGATTCCAGAGGC
TAG
CTCTCCTGGCCTCTCCCTCCC
AGVTQTPKFQV
TGGGGCCCAGGCAGGGAGA
LKTGQSMTLQC
ATGTCTCAGAATGACTTCCTT
AQDMNHEYMS
GAGAGTCCTGCTCCCCTTTC
WYRQDPGMGL
ATCAATGCACAGATACAGAA
RLIHYSVGAGIT
GACCCCTCCGTCATGCAGCA
DQGEVPNGYN
TCTGCCATGAGCATCGGCCT
VSRSTTEDFPLR
CCTGTGCTGTGCAGCCTTGTC
LLSAAPSQTSV
TCTCCTGTGGGCAGGTCCAG
YFCASSHLGAG
TGAATGCTGGTGTCACTCAG
GPHEQYFGPGT
RLTVTEDLKNV
FPPEVAVFEPS
EAEISHT
Q
KAT
LVCLATGFYP
DHVELSWWVN
GKEVHSGVST
DPCIPLKEQPA
LNDSRYCLSSR
LRVSATFWQN
PRNHFRCQVQ
FYGLSENDEW
TQDRAKPVTQI
VSAEAWGRAD
CGFTSESYQ
Q
GVLSATILYEI
LLGKATLYAV
LVSALVLMAM
ACGTGTTCCCACCCGAGGT
VKRKDSRG
CGCTGTGTTTGAGCCATCA
GAAGCAGAGATCTCCCACA
CCCAAAAGGCCACACTGGT
GTGCCTGGCCACAGGCTTC
TACCCCGACCACGTGGAGC
TGAGCTGGTGGGTGAATGG
GAAGGAGGTGCACAGTGG
GGTCAGCACAGACCCGCAG
CCCCTCAAGGAGCAGCCCG
CCCTCAATGACTCCAGATA
CTGCCTGAGCAGCCGCCTG
AGGGTCTCGGCCACCTTCT
GGCAGAACCCCCGCAACCA
CTTCCGCTGTCAAGTCCAG
TTCTACGGGCTCTCGGAGA
ATGACGAGTGGACCCAGGA
TAGGGCCAAACCTGTCACC
CAGATCGTCAGCGCCGAGG
CCTGGGGTAGAGCAGACTG
TGGCTTCACCTCCGAGTCT
TACCAGCAAGGGGTCCTGT
CTGCCACCATCCTCTATGA
GATCTTGCTAGGGAAGGCC
ACCTTGTATGCCGTGCTGG
TCAGTGCCCTCGTGCTGAT
GGCCATGGTCAAGAGAAAG
GATTCCAGAGGCTAG
GVSQSPRYKVT
KRGQDVALRC
DPISGHVSLYW
YRQALGQGPEF
LTYFNYEAQQD
KSGLPNDRFSA
ERPEGSISTLTIQ
RTEQRDSAMYR
CASSHTDGSYE
QYFGPGTRLTV
TEDLKNVFPPE
VAVFEPSEAEI
SHTQKATLVC
LATGFYPDHV
ELSWWVNGKE
VHSGVSTDPQP
LKEQPALNDS
RYCLSSRLRVS
ATFWQNPRNH
FRC
Q
V
Q
FYGL
SENDEWTQDR
GGGCACCAGTCTCCTATGCT
AKPVT
Q
IVSAE
GGGTGGTCCTGGGTTTCCTA
AWGRADCGFT
GGGACAGATCACACAGGTGC
SESYQQGVLSA
TILYEILLGKA
TLYAVLVSALV
LMAMVKRKDS
RG
ACCTGAAAAACGTGTTCCC
ACCCGAGGTCGCTGTGTTT
GAGCCATCAGAAGCAGAGA
TCTCCCACACCCAAAAGGC
CACACTGGTATGCCTGGCC
ACAGGCTTCTACCCCGACC
ACGTGGAGCTGAGCTGGTG
GGTGAATGGGAAGGAGGT
GCACAGTGGGGTCAGCACA
GACCCGCAGCCCCTCAAGG
AGCAGCCCGCCCTCAATGA
CTCCAGATACTGCCTGAGC
AGCCGCCTGAGGGTCTCGG
CCACCTTCTGGCAGAACCC
CCGCAACCACTTCCGCTGT
CAAGTCCAGTTCTACGGGC
TCTCGGAGAATGACGAGTG
GACCCAGGATAGGGCCAAA
CCTGTCACCCAGATCGTCA
GCGCCGAGGCCTGGGGTA
GAGCAGACTGTGGCTTCAC
CTCCGAGTCTTACCAGCAA
GGGGTCCTGTCTGCCACCA
TCCTCTATGAGATCTTGCT
AGGGAAGGCCACCTTGTAT
GCCGTGCTGGTCAGTGCCC
TCGTGCTGATGGCCATGGT
CAAGAGAAAGGATTCCAGA
GGCTAG
(SEQ ID NO: 295)
GITQSPKYLFRK
EGQNVTLSCEQ
NLNHDAMYWY
RQDPGQGLRLI
YYSQIVNDFQK
GDIAEGYSVSR
EKKESFPLTVTS
AQKNPTAFYLC
ASSIGAFAGQP
QHFGDGTRLSIL
AGGTGCTCTGCTGTGTGGTC
EDLNKVFPPEV
CTTTGTCTCCTGGGAGCAAA
AVFEPSEAEIS
CACCGTGGATGGTGGAATCA
HT
Q
KATLVCL
ATGFYPDHVE
LSWWVNGKE
VHSGVSTD
Q
P
LKE
Q
PALNDS
RYCL
CAAGGTGTTCCCACCCGAG
GTCGCTGTGTTTGAGCCAT
CAGAAGCAGAGATCTCCCA
CACCCAAAAGGCCACACTG
GTGTGCCTGGCCACAGGCT
TCTACCCCGACCACGTGGA
GCTGAGCTGGTGGGTGAAT
GGGAAGGAGGTGCACAGT
GGGGTCAGCACAGACCCG
CAGCCCCTCAAGGAGCAGC
CCGCCCTCAATGACTCCAG
ATACTGCCTGAGCAGCCGC
CTGAGGGTCTCGGCCACCT
TCTGGCAGAACCCCCGCAA
CCACTTCCGCTGTCAAGTC
CAGTTCTACGGGCTCTCGG
AGAATGACGAGTGGACCCA
GGATAGGGCCAAACCCGTC
ACCCAGATCGTCAGCGCCG
AGGCCTGGGGTAGAGCAG
ACTGTGGCTTTACCTCGGT
GTCCTACCAGCAAGGGGTC
CTGTCTGCCACCATCCTCT
ATGAGATCCTGCTAGGGAA
GGCCACCCTGTATGCTGTG
CTGGTCAGCGCCCTTGTGT
TGATGGCCATGGTCAAGAG
AAAGGATTTCTGA
GVSQNPRHKIT
KRGQNVTFRCD
PISEHNRLYWY
GACCCTGCCATGGGCACCAG
CCTCCTCTGCTGGATGGCCCT
TYFQNEAQLEK
GTGTCTCCTGGGGGCAGATC
SRLLSDRFSAER
ACGCAGATACTGGAGTCTCC
PKGSFSTLEIQR
TEQGDSAMYLC
ASSLAPTPGPDT
GELFFGEGSRLT
VLEDLKNVFPP
EVAVFEPSEAE
ISHT
Q
KATLVC
LATCFYPDHV
ELSWWVNGKE
VHSGVSTDPQP
LKE
Q
PALNDS
RYCLSSRLKVS
ATFWQNPRNH
KRCQVQFYGL
SENDEWTQDR
AKPVTQIVSAE
AWGRADCGFT
SESYQQGVLSA
TILYEILLGKA
TCCCACCCGAGGTCGCTGT
TLYAVLVSALV
GTTTGAGCCATCAGAAGCA
LMAMVKRKDS
GAGATCTCCCACACCCAAA
RG
AGGCCACACTGGTGTGCCT
GGCCACAGGCTTCTACCCC
GACCACGTGGAGCTGAGCT
GGTGGGTGAATGGGAAGG
AGGTGCACAGTGGGGTCA
GCACAGACCCGCAGCCCCT
CAAGGAGCAGCCCGCCCTC
AATGACTCCAGATACTGCC
TGAGCAGCCGCCTGAGGGT
CTCGGCCACCTTCTGGCAG
AACCCCCGCAACCACTTCC
GCTGTCAAGTCCAGTTCTA
CGGGCTCTCGGAGAATGAC
GAGTGGACCCAGGATAGG
GCCAAACCTGTCACCCAGA
TCGTCAGCGCCGAGGCCTG
GGGTAGAGCAGACTGTGG
CTTCACCTCCGAGTCTTAC
CAGCAAGGGGTCCTGTCTG
CCACCATCCTCTATGAGAT
CTTGCTAGGGAAGGCCACC
TTGTATGCCGTGCTGGTCA
GTGCCCTCGTGCTGATGGC
CATGGTCAAGAGAAAGGAT
TCCAGAGGCTAG
(SEQ ID
QVTQNPRYLIT
VTGKKLTVTCS
QNMNHEYMSW
YRQDPGLGLRQ
IYYSMNVEVTD
KGDVPEGYKVS
TCCTTGGCTATGTGGTCCTTT
RKEKRNFPLILE
GCCTTCTAGGAGCAGGCCCC
SPSPNQTSLYFC
CTGGAAGCCCAAGTGACCCA
ASSLGNIYEQYF
GPGTRLTVTED
LKNVFPPEVAV
FEPSEAEISHT
QKATLVCLAT
GFYPDHVELS
WWVNGKEVH
SGVSTDP
Q
PLK
EQPALNDSRY
CLSSRLRVSAT
FWQNPRNHFR
CQVQFYGLSE
NDEWTQDRAK
PVTQIVSAEAW
GRADCGFTSES
YQQGVLSATIL
YEILLGKATLY
AVLVSALVLM
CCCACCCGAGGTCGCTGTG
AMVKRKDSRG
TTTGAGCCATCAGAAGCAG
AGATCTCCCACACCCAAAA
GGCCACACTGGTGTGCCTG
GCCACAGGCTTCTACCCCG
ACCACGTGGAGCTGAGCTG
GTGGGTGAATGGGAAGGA
GGTGCACAGTGGGGTCAG
CACAGACCCGCAGCCCCTC
AAGGAGCAGCCCGCCCTCA
ATGACTCCAGATACTGCCT
GAGCAGCCGCCTGAGGGT
CTCGGCCACCTTCTGGCAG
AACCCCCGCAACCACTTCC
GCTGTCAAGTCCAGTTCTA
CGGGCTCTCGGAGAATGAC
GAGTGGACCCAGGATAGG
GCCAAACCTGTCACCCAGA
TCGTCAGCGCCGAGGCCTG
GGGTAGAGCAGACTGTGG
CTTCACCTCCGAGTCTTAC
CAGCAAGGGGTCCTGTCTG
CCACCATCCTCTATGAGAT
CTTGCTAGGGAAGGCCACC
TTGTATGCCGTGCTGGTCA
GTGCCCTCGTGCTGATGGC
CATGGTCAAGAGAAAGGAT
TCCAGAGGCTAG
(SEQ ID
GVSQTPSNKVT
EKGKYVELRCD
PISGHTALYWY
RQSLGQGPEFLI
ATGGGCACCAGGCTCCTCTG
YFQGTGAADDS
CTGGGCAGCCCTGTGCCTCC
GLPNDRFFAVR
TGGGGGCAGATCACACAGGT
PEGSVSTLKIQR
TERGDSAVYLC
ASSLGSGEQFF
GPGTRLTVLED
LKNVFPPEVAV
FEPSEAEISHT
QKATLVCLAT
GFYPDHVELS
WWVNGKEVH
SGVSTDPQPLK
EQPALNDSRY
CLSSRLRVSAT
FWQNPRNHER
CQVQFYGLSE
NDEWTQDRAK
PVTQIVSAEAW
GRADCGFTSES
YQQGVLSATIL
GAAAAACGTGTTCCCACCC
YEILLGKATLY
GAGGTCGCTGTGTTTGAGC
AVLVSALVLM
CATCAGAAGCAGAGATCTC
AMVKRKDSRG
CCACACCCAAAAGGCCACA
CTGGTGTGCCTGGCCACAG
GCTTCTACCCCGACCACGT
GGAGCTGAGCTGGTGGGT
GAATGGGAAGGAGGTGCA
CAGTGGGGTCAGCACAGAC
CCGCAGCCCCTCAAGGAGC
AGCCCGCCCTCAATGACTC
CAGATACTGCCTGAGCAGC
CGCCTGAGGGTCTCGGCCA
CCTTCTGGCAGAACCCCCG
CAACCACTTCCGCTGTCAA
GTCCAGTTCTACGGGCTCT
CGGAGAATGACGAGTGGA
CCCAGGATAGGGCCAAACC
TGTCACCCAGATCGTCAGC
GCCGAGGCCTGGGGTAGA
GCAGACTGTGGCTTCACCT
CCGAGTCTTACCAGCAAGG
GGTCCTGTCTGCCACCATC
CTCTATGAGATCTTGCTAG
GGAAGGCCACCTTGTATGC
CGTGCTGGTCAGTGCCCTC
GTGCTGATGGCCATGGTCA
AGAGAAAGGATTCCAGAGG
CTAG
VTQSPTHLIKTR
GQQATLRCSPIS
GHTSVYWYQQ
ALGLGLQFLLW
YDEGEERNRGN
FPPRFSGRQFPN
YSSELNVNALE
LEDSALYLCAS
SLQTSYEQYFG
PGTRLTVTEDL
KNVFPPEVAVF
EPSEAEISHTQ
KATLVCLATG
GGGACCCAGGCTCCTCTTCT
FYPDHVELSW
GGGCACTGCTTTGTCTCCTCG
WVNGKEVHSG
GAACAGGCCCAGTGGAGGCT
VSTDPQPLKEQ
PALNDSRYCLS
SRLRVSATFW
QNPRNHFRCQ
VQFYGLSEND
EWTQDRAKPV
TQIVSAEAWG
RADCGFTSESY
QQGVLSATILY
EILLGKATLYA
VLVSALVLMA
MVKRKDSRG
AAAACGTGTTCCCACCCGA
GGTCGCTGTGTTTGAGCCA
TCAGAAGCAGAGATCTCCC
ACACCCAAAAGGCCACACT
GGTATGCCTGGCCACAGGC
TTCTACCCCGACCACGTGG
AGCTGAGCTGGTGGGTGAA
TGGGAAGGAGGTGCACAG
TGGGGTCAGCACAGACCCG
CAGCCCCTCAAGGAGCAGC
CCGCCCTCAATGACTCCAG
ATACTGCCTGAGCAGCCGC
CTGAGGGTCTCGGCCACCT
TCTGGCAGAACCCCCGCAA
CCACTTCCGCTGTCAAGTC
CAGTTCTACGGGCTCTCGG
AGAATGACGAGTGGACCCA
GGATAGGGCCAAACCTGTC
ACCCAGATCGTCAGCGCCG
AGGCCTGGGGTAGAGCAG
ACTGTGGCTTCACCTCCGA
GTCTTACCAGCAAGGGGTC
CTGTCTGCCACCATCCTCT
ATGAGATCTTGCTAGGGAA
GGCCACCTTGTATGCCGTG
CTGGTCAGTGCCCTCGTGC
TGATGGCCATGGTCAAGAG
AAAGGATTCCAGAGGCTAG
QVTQNPRYLIT
VTGKKLTVTCS
QNMNHEYMSW
YRQDPGLGLRQ
IYYSMNVEVTD
KGDVPEGYKVS
RKEKRNFPLILE
CAGCTCCTTGGCTATGTGGT
SPSPNQTSLYFC
CCTTTGCCTTCTAGGAGCAG
ASSLSWGTGKR
GCCCCCTGGAAGCCCAAGTG
ADTQYFGPGTR
LTVLEDLKNVF
PPEVAVFEPSE
AEISHTQKATL
VCLATGFYPD
HVELSWWVN
GKEVHSGVST
DPQPLKEQPA
LNDSRYCLSSR
LRVSATFWQN
PRNHFRCQVQ
FYGLSENDEW
TQDRAKPVTQI
VSAEAWGRAD
CGFTSESYQQ
GVLSATILYEI
LLGKATLYAV
LVSALVLMAM
VKRKDSRG
CCCACCCGAGGTCGCTGTG
TTTGAGCCATCAGAAGCAG
AGATCTCCCACACCCAAAA
GGCCACACTGGTGTGCCTG
GCCACAGGCTTCTACCCCG
ACCACGTGGAGCTGAGCTG
GTGGGTGAATGGGAAGGA
GGTGCACAGTGGGGTCAG
CACAGACCCGCAGCCCCTC
AAGGAGCAGCCCGCCCTCA
ATGACTCCAGATACTGCCT
GAGCAGCCGCCTGAGGGT
CTCGGCCACCTTCTGGCAG
AACCCCCGCAACCACTTCC
GCTGTCAAGTCCAGTTCTA
CGGGCTCTCGGAGAATGAC
GAGTGGACCCAGGATAGG
GCCAAACCTGTCACCCAGA
TCGTCAGCGCCGAGGCCTG
GGGTAGAGCAGACTGTGG
CTTCACCTCCGAGTCTTAC
CAGCAAGGGGTCCTGTCTG
CCACCATCCTCTATGAGAT
CTTGCTAGGGAAGGCCACC
TTGTATGCCGTGCTGGTCA
GTGCCCTCGTGCTGATGGC
CATGGTCAAGAGAAAGGAT
TCCAGAGGCTAG
(SEQ ID
GVSQDPRHKIT
KRGQNVTFRCD
PISEHNRLYWY
RQTLGQGPEFL
TYFQNEAQLEK
SRLLSDRFSAER
PKGSFSTLEIQR
TEQGDSAMYLC
ASSLVGGGSNE
QFFGPGTRLTV
LEDLKNVFPPE
VAVFEPSEAEI
SHTQKATLVC
LATGFYPDHV
ELSWWVNGKE
VHSGVSTDPQP
LKEQPALNDS
GGCACCAGCCTCCTCTGCTG
RYCLSSRLRVS
GATGGCCCTGTGTCTCCTGG
ATFWQNPRNH
GGGCAGATCACGCAGATACT
FRCQVQFYGL
SENDEWTQDR
AKPVTQIVSAE
AWGRADCGFT
SESYQQGVLSA
TILYEILLGKA
TLYAVLVSALV
LMAMVKRKDS
RG
CCCACCCGAGGTCGCTGTG
TTTGAGCCATCAGAAGCAG
AGATCTCCCACACCCAAAA
GGCCACACTGGTGTGCCTG
GCCACAGGCTTCTACCCCG
ACCACGTGGAGCTGAGCTG
GTGGGTGAATGGGAAGGA
GGTGCACAGTGGGGTCAG
CACAGACCCGCAGCCCCTC
AAGGAGCAGCCCGCCCTCA
ATGACTCCAGATACTGCCT
GAGCAGCCGCCTGAGGGT
CTCGGCCACCTTCTGGCAG
AACCCCCGCAACCACTTCC
GCTGTCAAGTCCAGTTCTA
CGGGCTCTCGGAGAATGAC
GAGTGGACCCAGGATAGG
GCCAAACCTGTCACCCAGA
TCGTCAGCGCCGAGGCCTG
GGGTAGAGCAGACTGTGG
CTTCACCTCCGAGTCTTAC
CAGCAAGGGGTCCTGTCTG
CCACCATCCTCTATGAGAT
CTTGCTAGGGAAGGCCACC
TTGTATGCCGTGCTGGTCA
GTGCCCTCGTGCTGATGGC
CATGGTCAAGAGAAAGGAT
TCCAGAGGCTAG
GVSQDPRHKIT
KRGQNVTFRCD
PISEHNRLYWY
RQTLGQGPEFL
TYFQNEAQLEK
SRLLSDRFSAER
PKGSFSTLEIQR
TEQGDSAMYLC
ASSLVGGTEAF
FGQGTRLTVVE
DLNKVFPPEVA
VFEPSEAEISH
TQKATLVCLA
TGFYPDHVELS
WWVNGKEVH
SGVSTDPQPLK
EQPALNDSRY
CLSSRLRVSAT
FWQNPRNHFR
CQVQFYGLSE
NDEWTQDRAK
PVTQIVSAEAW
GRADCGFTSVS
YQQGVLSATIL
YEILLGKATLY
AVLVSALVLM
GGCACCAGCCTCCTCTGCTG
AMVKRKDF
GATGGCCCTGTGTCTCCTGG
GGGCAGATCACGCAGATACT
TGAACAAGGTGTTCCCACC
CGAGGTCGCTGTGTTTGAG
CCATCAGAAGCAGAGATCT
CCCACACCCAAAAGGCCAC
ACTGGTGTGCCTGGCCACA
GGCTTCTACCCCGACCACG
TGGAGCTGAGCTGGTGGGT
GAATGGGAAGGAGGTGCA
CAGTGGGGTCAGCACAGAC
CCGCAGCCCCTCAAGGAGC
AGCCCGCCCTCAATGACTC
CAGATACTGCCTGAGCAGC
CGCCTGAGGGTCTCGGCCA
CCTTCTGGCAGAACCCCCG
CAACCACTTCCGCTGTCAA
GTCCAGTTCTACGGGCTCT
CGGAGAATGACGAGTGGA
CCCAGGATAGGGCCAAACC
CGTCACCCAGATCGTCAGC
GCCGAGGCCTGGGGTAGA
GCAGACTGTGGCTTTACCT
CGGTGTCCTACCAGCAAGG
GGTCCTGTCTGCCACCATC
CTCTATGAGATCCTGCTAG
GGAAGGCCACCCTGTATGC
TGTGCTGGTCAGCGCCCTT
GTGTTGATGGCCATGGTCA
AGAGAAAGGATTTCTGA
GVSQDPRHKIT
KRGQNVTFRCD
PISEHNRLYWY
RQTLGQGPEFL
TYFQNEAQLEK
SRLLSDRFSAER
GGGCACCAGCCTCCTCTGCT
PKGSFSTLEIQR
GGATGGCCCTGTGTCTCCTG
TEQGDSAMYLC
GGGGCAGATCACGCAGATAC
ASSPDRNLGQY
FGPGTRLTVTE
DLKNVFPPEVA
VFEPSEAEISH
TQKATLVCLA
TGFYPDHVELS
WWVNGKEVH
SGVSTDPQPLK
EQPALNDSRY
CLSSRLRVSAT
FWQNPRNHFR
CQVQFYGLSE
NDEWTQDRAK
PVTQIVSAEAW
GRADCGFTSES
YQQGVLSATIL
YEILLGKATLY
AVLVSALVLM
CCTGAAAAACGTGTTCCCA
AMVKRKDSRG
CCCGAGGTCGCTGTGTTTG
AGCCATCAGAAGCAGAGAT
CTCCCACACCCAAAAGGCC
ACACTGGTGTGCCTGGCCA
CAGGCTTCTACCCCGACCA
CGTGGAGCTGAGCTGGTG
GGTGAATGGGAAGGAGGT
GCACAGTGGGGTCAGCACA
GACCCGCAGCCCCTCAAGG
AGCAGCCCGCCCTCAATGA
CTCCAGATACTGCCTGAGC
AGCCGCCTGAGGGTCTCGG
CCACCTTCTGGCAGAACCC
CCGCAACCACTTCCGCTGT
CAAGTCCAGTTCTACGGGC
TCTCGGAGAATGACGAGTG
GACCCAGGATAGGGCCAAA
CCTGTCACCCAGATCGTCA
GCGCCGAGGCCTGGGGTA
GAGCAGACTGTGGCTTCAC
CTCCGAGTCTTACCAGCAA
GGGGTCCTGTCTGCCACCA
TCCTCTATGAGATCTTGCT
AGGGAAGGCCACCTTGTAT
GCCGTGCTGGTCAGTGCCC
TCGTGCTGATGGCCATGGT
CAAGAGAAAGGATTCCAGA
GGCTAG
(SEQ ID NO: 358)
VTQFPSHSVIEK
QTVTLRCDPIS
GHDNLYWYRR
VMGKEIKFLLH
FVKESKQDESG
CTCAGTTTAGTGTCCCTTTGT
MPNNRFLAERT
CTCCTGGGAGCAAAGCACAT
GGTYSTLKVQP
AGAAGCTGGAGTTACTCAGT
AELEDSGVYFC
ASSQVQAFNEQ
FFGPGTRLTVL
EDLKNVFPPEV
AVFEPSEAEIS
HTQKATLVCL
ATGFYPDHVE
LSWWVNGKE
VHSGVSTDPQP
LKEQPALNDS
RYCLSSRLRVS
ATFWQNPRNH
FRCQVQFYGL
SENDEWTQDR
AKPVTQIVSAE
AWGRADCGFT
SESYQQGVLSA
TILYEILLGKA
TTCCCACCCGAGGTCGCTG
TLYAVLVSALV
TGTTTGAGCCATCAGAAGC
LMAMVKRKDS
AGAGATCTCCCACACCCAA
RG
AAGGCCACACTGGTGTGCC
TGGCCACAGGCTTCTACCC
CGACCACGTGGAGCTGAGC
TGGTGGGTGAATGGGAAG
GAGGTGCACAGTGGGGTC
AGCACAGACCCGCAGCCCC
TCAAGGAGCAGCCCGCCCT
CAATGACTCCAGATACTGC
CTGAGCAGCCGCCTGAGG
GTCTCGGCCACCTTCTGGC
AGAACCCCCGCAACCACTT
CCGCTGTCAAGTCCAGTTC
TACGGGCTCTCGGAGAATG
ACGAGTGGACCCAGGATAG
GGCCAAACCTGTCACCCAG
ATCGTCAGCGCCGAGGCCT
GGGGTAGAGCAGACTGTG
GCTTCACCTCCGAGTCTTA
CCAGCAAGGGGTCCTGTCT
GCCACCATCCTCTATGAGA
TCTTGCTAGGGAAGGCCAC
CTTGTATGCCGTGCTGGTC
AGTGCCCTCGTGCTGATGG
CCATGGTCAAGAGAAAGGA
TTCCAGAGGCTAG
(SEQ ID
PRYKIIEKRQSV
AFWCNPISGHA
TLYWYQQILGQ
GPKLLIQFQNN
GVVDDSQLPKD
CGTCCCTCCTCCTCTGCTTTT
RFSAERLKGVD
GCTCACAGTGACCCTGATTG
STLKIQPAKLED
GGCAAAGCTCCCATCCTTCC
SAVYLCASSRG
CTGACCCTGCCATGGGCACC
EPGSGANVLTF
AGGCTCCTCTGCTGGGCGGC
GAGSRLTVLED
CCTCTGTCTCCTGGGAGCAG
LKNVFPPEVAV
AACTCACAGAAGCTGGAGTT
FEPSEAEISHT
QKATLVCLAT
GFYPDHVELS
WWVNGKEVH
SGVSTDPQPLK
EQPALNDSRY
CLSSRLRVSAT
FWQNPRNHFR
CQVQFYGLSE
NDEWTQDRAK
PVTQIVSAEAW
GRADCGFTSES
YQQGVLSATIL
YEILLGKATLY
AVLVSALVLM
AMVKRKDSRG
CCACCCGAGGTCGCTGTGT
TTGAGCCATCAGAAGCAGA
GATCTCCCACACCCAAAAG
GCCACACTGGTGTGCCTGG
CCACAGGCTTCTACCCCGA
CCACGTGGAGCTGAGCTGG
TGGGTGAATGGGAAGGAG
GTGCACAGTGGGGTCAGCA
CAGACCCGCAGCCCCTCAA
GGAGCAGCCCGCCCTCAAT
GACTCCAGATACTGCCTGA
GCAGCCGCCTGAGGGTCTC
GGCCACCTTCTGGCAGAAC
CCCCGCAACCACTTCCGCT
GTCAAGTCCAGTTCTACGG
GCTCTCGGAGAATGACGAG
TGGACCCAGGATAGGGCCA
AACCTGTCACCCAGATCGT
CAGCGCCGAGGCCTGGGG
TAGAGCAGACTGTGGCTTC
ACCTCCGAGTCTTACCAGC
AAGGGGTCCTGTCTGCCAC
CATCCTCTATGAGATCTTG
CTAGGGAAGGCCACCTTGT
ATGCCGTGCTGGTCAGTGC
CCTCGTGCTGATGGCCATG
GTCAAGAGAAAGGATTCCA
GAGGCTAG
(SEQ ID NO: 372)
VTQSPTHLIKTR
GQQVTLRCSPK
SGHDTVSWYQ
QALGQGPQFIF
QYYEEEERQRG
NFPDRFSGHQF
PNYSSELNVNA
LLLGDSALYLC
ASSSGTDPSGA
CTTCCTGTCCCTCTGTGCCAT
NVLTFGAGSRL
GGGCCCCGGGCTCCTCTGCT
TVLEDLKNVFP
GGGCACTGCTTTGTCTCCTG
PEVAVFEPSEA
GGAGCAGGCTTAGTGGACGC
EISHTQKATLV
CLATGFFPDH
VELSWWVNG
KEVHSGVSTDP
QPLKEQPALN
DSRYCLSSRLR
VSATFWQNPR
NHFRCQVQFY
GLSENDEWTQ
DRAKPVTQIVS
AEAWGRADCG
FTSESYQQGVL
SATILYEILLG
KATLYAVLVS
ALVLMAMVK
RKDSRG
TGTTCCCACCCGAGGTCGC
TGTGTTTGAGCCATCAGAA
GCAGAGATCTCCCACACCC
AAAAGGCCACACTGGTGTG
CCTGGCCACAGGCTTCTTC
CCTGACCACGTGGAGCTGA
GCTGGTGGGTGAATGGGA
AGGAGGTGCACAGTGGGG
TCAGCACAGACCCGCAGCC
CCTCAAGGAGCAGCCCGCC
CTCAATGACTCCAGATACT
GCCTGAGCAGCCGCCTGAG
GGTCTCGGCCACCTTCTGG
CAGAACCCCCGCAACCACT
TCCGCTGTCAAGTCCAGTT
CTACGGGCTCTCGGAGAAT
GACGAGTGGACCCAGGATA
GGGCCAAACCTGTCACCCA
GATCGTCAGCGCCGAGGCC
TGGGGTAGAGCAGACTGTG
GCTTCACCTCCGAGTCTTA
CCAGCAAGGGGTCCTGTCT
GCCACCATCCTCTATGAGA
TCTTGCTAGGGAAGGCCAC
CTTGTATGCCGTGCTGGTC
AGTGCCCTCGTGCTGATGG
CCATGGTCAAGAGAAAGGA
TTCCAGAGGCTAG
(SEQ ID
QVTQNPRYLIT
VTGKKLTVTCS
QNMNHEYMSW
YRQDPGLGLRQ
IYYSMNVEVTD
KGDVPEGYKVS
TCCTTGGCTATGTGGTCCTTT
RKEKRNFPLILE
GCCTTCTAGGAGCAGGCCCC
SPSPNQTSLYFC
CTGGAAGCCCAAGTGACCCA
ASSTGQNIGGE
LFFGEGSRLTVL
EDLKNVFPPEV
AVFEPSEAEIS
HTQKATLVCL
ATGFYPDHVE
LSWWVNGKE
VHSGVSTDPQP
LKEQPALNDS
RYCLSSRLRVS
ATFWQNPRNH
FRCQVQFYGL
SENDEWTQDR
AKPVTQIVSAE
AWGRADCGFT
SESYQQGVLSA
TILYEILLGKA
TLYAVLVSALV
CGTGTTCCCACCCGAGGTC
LMAMVKRKDS
GCTGTGTTTGAGCCATCAG
RG
AAGCAGAGATCTCCCACAC
CCAAAAGGCCACACTGGTG
TGCCTGGCCACAGGCTTCT
ACCCCGACCACGTGGAGCT
GAGCTGGTGGGTGAATGG
GAAGGAGGTGCACAGTGG
GGTCAGCACAGACCCGCAG
CCCCTCAAGGAGCAGCCCG
CCCTCAATGACTCCAGATA
CTGCCTGAGCAGCCGCCTG
AGGGTCTCGGCCACCTTCT
GGCAGAACCCCCGCAACCA
CTTCCGCTGTCAAGTCCAG
TTCTACGGGCTCTCGGAGA
ATGACGAGTGGACCCAGGA
TAGGGCCAAACCTGTCACC
CAGATCGTCAGCGCCGAGG
CCTGGGGTAGAGCAGACTG
TGGCTTCACCTCCGAGTCT
TACCAGCAAGGGGTCCTGT
CTGCCACCATCCTCTATGA
GATCTTGCTAGGGAAGGCC
ACCTTGTATGCCGTGCTGG
TCAGTGCCCTCGTGCTGAT
GGCCATGGTCAAGAGAAAG
GATTCCAGAGGCTAG
(SEQ
ACATCAGAGTTGCTGCCATC
TTAGACTTAACTCATCAGTA
TCAGGTGATCCTGAGGCTCA
RHKITKRGQNV
GTGATGTCACTGTGGGAACT
TFRCDPISEHNR
GCTCTGTGGCGACAAGGACG
LYWYRQTLGQ
TCCCTCATCCTCTGCTCCTGC
GPEFLTYFQNE
TCACAGTGACCCTGATCTGG
AQLEKSRLLSD
TAAAGCTCCCATCCTGCCCT
RFSAERPKGSFS
GACCCTGCCATGGGCACCAG
TLEIQRTEQGDS
CCTCCTCTGCTGGATGGCCCT
AMYLCASSTHE
GTGTCTCCTGGGGGCAGATC
KTGWKSPLHFG
ACGCAGATACTGGAGTCTCC
NGTRLTVTEDL
NKVFPPEVAVF
EPSEAEISHTQ
KATLVCLATG
FYPDHVELSW
WVNGKEVHSG
VSTDPQPLKEQ
PALNDSRYCLS
SRLRVSATFW
QNPRNHFRCQ
VQFYGLSEND
EWTQDRAKPV
TQIVSAEAWG
RADCGFTSVSY
QQGVLSATILY
EILLGKATLYA
VLVSALVLMA
MVKRKDF
GACCTGAACAAGGTGTTCC
CACCCGAGGTCGCTGTGTT
TGAGCCATCAGAAGCAGAG
ATCTCCCACACCCAAAAGG
CCACACTGGTGTGCCTGGC
CACAGGCTTCTACCCCGAC
CACGTGGAGCTGAGCTGGT
GGGTGAATGGGAAGGAGG
TGCACAGTGGGGTCAGCAC
AGACCCGCAGCCCCTCAAG
GAGCAGCCCGCCCTCAATG
ACTCCAGATACTGCCTGAG
CAGCCGCCTGAGGGTCTCG
GCCACCTTCTGGCAGAACC
CCCGCAACCACTTCCGCTG
TCAAGTCCAGTTCTACGGG
CTCTCGGAGAATGACGAGT
GGACCCAGGATAGGGCCA
AACCCGTCACCCAGATCGT
CAGCGCCGAGGCCTGGGG
TAGAGCAGACTGTGGCTTT
ACCTCGGTGTCCTACCAGC
AAGGGGTCCTGTCTGCCAC
CATCCTCTATGAGATCCTG
CTAGGGAAGGCCACCCTGT
ATGCTGTGCTGGTCAGCGC
CCTTGTGTTGATGGCCATG
GTCAAGAGAAAGGATTTCT
GA
(SEQ ID NO: 393)
MGQEVILRCVPI
SNHLYFYWYR
QILGQKVEFLV
CTCGTATGCTGGGCAATTTTT
SFYNNEISEKSE
AGTCTCTTGAAAGCAGGACT
IFDDQFSVERPD
CACAGAACCTGAAGTCACCC
GSNFTLKIRSTK
LEDSAMYFCAS
STHSDRNLNTE
AFFGQGTRLTV
VEDLNKVFPPE
VAVFEPSEAEI
SHTMATLVC
LATGFFPDHVE
LSWWVNGKE
VHSGVSTDPQP
LKEQPALNDS
RYCLSSRLRVS
ATFWQNPRNH
FRCQVQFYGL
SENDEWTQDR
AKPVTQIVSAE
AWGRADCGFT
SVSYQQGVLSA
GAACAAGGTGTTCCCACCC
TILYEILLGKA
GAGGTCGCTGTGTTTGAGC
TLYAVLVSALV
CATCAGAAGCAGAGATCTC
LMAMVKRKD
CCACACCCAAAAGGCCACA
F
CTGGTGTGCCTGGCCACAG
GCTTCTTCCCTGACCACGT
GGAGCTGAGCTGGTGGGT
GAATGGGAAGGAGGTGCA
CAGTGGGGTCAGCACAGAC
CCGCAGCCCCTCAAGGAGC
AGCCCGCCCTCAATGACTC
CAGATACTGCCTGAGCAGC
CGCCTGAGGGTCTCGGCCA
CCTTCTGGCAGAACCCCCG
CAACCACTTCCGCTGTCAA
GTCCAGTTCTACGGGCTCT
CGGAGAATGACGAGTGGA
CCCAGGATAGGGCCAAACC
CGTCACCCAGATCGTCAGC
GCCGAGGCCTGGGGTAGA
GCAGACTGTGGCTTTACCT
CGGTGTCCTACCAGCAAGG
GGTCCTGTCTGCCACCATC
CTCTATGAGATCCTGCTAG
GGAAGGCCACCCTGTATGC
TGTGCTGGTCAGCGCCCTT
GTGTTGATGGCCATGGTCA
AGAGAAAGGATTTCTGA
VTQTPSHQVTQ
MGQEVILRCVPI
SNHLYFYWYR
QILGQKVEFLV
GTATGCTGGGCAATTTTTAG
SFYNNEISEKSE
TCTCTTGAAAGCAGGACTCA
IFDDQFSVERPD
CAGAACCTGAAGTCACCCAG
GSNFTLKIRSTK
LEDSAMYFCAS
SVQATGHGYTF
GSGTRLTVVED
LNKVFPPEVAV
FEPSEAEISHT
QKATLVCLAT
GFYPDHVELS
WWVNGKEVH
SGVSTDPQPLK
EQPALNDSRY
CLSSRLRVSAT
FWQNPRNHFR
CQVQFYGLSE
NDEWTQDRAK
PVTQIVSAEAW
GRADCGFTSVS
YQQGVLSATIL
TTCCCACCCGAGGTCGCTG
YEILLGKATLY
TGTTTGAGCCATCAGAAGC
AVLVSALVLM
AGAGATCTCCCACACCCAA
AMVKRKDF
AAGGCCACACTGGTATGCC
TGGCCACAGGCTTCTACCC
CGACCACGTGGAGCTGAGC
TGGTGGGTGAATGGGAAG
GAGGTGCACAGTGGGGTC
AGCACAGACCCGCAGCCCC
TCAAGGAGCAGCCCGCCCT
CAATGACTCCAGATACTGC
CTGAGCAGCCGCCTGAGG
GTCTCGGCCACCTTCTGGC
AGAACCCCCGCAACCACTT
CCGCTGTCAAGTCCAGTTC
TACGGGCTCTCGGAGAATG
ACGAGTGGACCCAGGATAG
GGCCAAACCCGTCACCCAG
ATCGTCAGCGCCGAGGCCT
GGGGTAGAGCAGACTGTG
GCTTTACCTCGGTGTCCTA
CCAGCAAGGGGTCCTGTCT
GCCACCATCCTCTATGAGA
TCCTGCTAGGGAAGGCCAC
CCTGTATGCTGTGCTGGTC
AGCGCCCTTGTGTTGATGG
CCATGGTCAAGAGAAAGGA
TTTCTGA
(SEQ ID NO: 407)
GVSQSPRYKVA
KRGQDVALRC
DPISGHVSLFW
YQQALGQGPEF
CTCCTCTGCTGGGTGGTCCTG
LTYFQNEAQLD
GGTTTCCTAGGGACAGATCA
KSGLPSDRFFAE
CACAGGTGCTGGAGTCTCCC
RPEGSVSTLKIQ
RTQQEDSAVYL
CASTPSGYNSW
EQFFGPGTRLT
VLEDLKNVFPP
EVAVFEPSEAE
ISHTQKATLVC
LATGFYPDHV
ELSWWVNGKE
VHSGVSTDP
Q
P
LKEQPALNDS
RYCLSSRLRVS
ATFWQNPRNH
FRCQVQFYGL
SENDEWT
Q
DR
AKPVTQIV
SAEAWGRADC
GFTSVSYQQG
AAACGTGTTCCCACCCGAG
VLSATILYEILL
GTCGCTGTGTTTGAGCCAT
GKATLYAVLV
CAGAAGCAGAGATCTCCCA
SALVLMAMVK
CACCCAAAAGGCCACACTG
RKDF
GTGTGCCTGGCCACAGGCT
TCTACCCCGACCACGTGGA
GCTGAGCTGGTGGGTGAAT
GGGAAGGAGGTGCACAGT
GGGGTCAGCACAGACCCG
CAGCCCCTCAAGGAGCAGC
CCGCCCTCAATGACTCCAG
ATACTGCCTGAGCAGCCGC
CTGAGGGTCTCGGCCACCT
TCTGGCAGAACCCCCGCAA
CCACTTCCGCTGTCAAGTC
CAGTTCTACGGGCTCTCGG
AGAATGACGAGTGGACCCA
GGATAGGGCCAAACCCGTC
ACCCAGATCGTCAGCGCCG
AGGCCTGGGGTAGAGCAG
ACTGTGGCTTTACCTCGGT
GTCCTACCAGCAAGGGGTC
CTGTCTGCCACCATCCTCT
ATGAGATCCTGCTAGGGAA
GGCCACCCTGTATGCTGTG
CTGGTCAGCGCCCTTGTGT
TGATGGCCATGGTCAAGAG
AAAGGATTTCTGA
(SEQ ID
QVTQNPRYLIT
VTGKKLTVTCS
QNMNHEYMSW
YRQDPGLGLRQ
IYYSMNVEVTD
KGDVPEGYKVS
CAGCTCCTTGGCTATGTGGT
RKEKRNFPLILE
CCTTTGCCTTCTAGGAGCAG
SPSPNQTSLYFC
GCCCCCTGGAAGCCCAAGTG
ATAPRGSNQPQ
HFGDGTRLSILE
DLNKVFPPEVA
VFEPSEAEISH
TQKATLVCLA
TGFYPDHVELS
WWVNGKEVH
SGVSTDPQPLK
EQPALNDSRY
CLSSRLRVSAT
FWQNPRNHFR
CQVQFYGLSE
NDEWTQDRAK
PVTQIVSAEAW
GRADCGFTSVS
Y
QQ
GVLSATIL
YEILLGKATLY
AVLVSALVLM
AGGTGTTCCCACCCGAGGT
AMVKRKDF
CGCTGTGTTTGAGCCATCA
GAAGCAGAGATCTCCCACA
CCCAAAAGGCCACACTGGT
GTGCCTGGCCACAGGCTTC
TACCCCGACCACGTGGAGC
TGAGCTGGTGGGTGAATGG
GAAGGAGGTGCACAGTGG
GGTCAGCACAGACCCGCAG
CCCCTCAAGGAGCAGCCCG
CCCTCAATGACTCCAGATA
CTGCCTGAGCAGCCGCCTG
AGGGTCTCGGCCACCTTCT
GGCAGAACCCCCGCAACCA
CTTCCGCTGTCAAGTCCAG
TTCTACGGGCTCTCGGAGA
ATGACGAGTGGACCCAGGA
TAGGGCCAAACCCGTCACC
CAGATCGTCAGCGCCGAGG
CCTGGGGTAGAGCAGACTG
TGGCTTTACCTCGGTGTCC
TACCAGCAAGGGGTCCTGT
CTGCCACCATCCTCTATGA
GATCCTGCTAGGGAAGGCC
ACCCTGTATGCTGTGCTGG
TCAGCGCCCTTGTGTTGAT
GGCCATGGTCAAGAGAAAG
GATTTCTGA
(SEQ ID NO:
ATGGAGGCAGTGGTCACAAC
TCTCCCCAGAGAAGGTGGTG
GAVVSQHPSW
TGAGGCCATCACGGAAGATG
VICKSGTSVKIE
CTGCTGCTTCTGCTGCTTCTG
CRSLDFQATTM
GGGCCAGGCTCCGGGCTTGG
FWYRQFPKQSL
MLMATSNEGS
KATYEQGVEK
DKFLINHASLTL
STLTVTSAHPE
DSSFYICRAWG
GSSYNEQFFGP
GTRLTVLEDLK
NVFPPEVAVFE
PSEAEISHTQK
ATLVCLATGF
YPDHVELSWW
VNGKEVHSGV
STDPQPLKEQP
ALNDSRYCLSS
RLRVSATFWQ
NPRNHFRC
Q
V
QFYGLSENDE
WTQDRAKPVT
CGTGTTCCCACCCGAGGTC
QIVSAEAWGR
GCTGTGTTTGAGCCATCAG
ADCGFTSVSYQ
AAGCAGAGATCTCCCACAC
QGVLSATILYE
CCAAAAGGCCACACTGGTA
ILLGKATLYAV
TGCCTGGCCACAGGCTTCT
LVSALVLMAM
ACCCCGACCACGTGGAGCT
VKRKDF
GAGCTGGTGGGTGAATGG
GAAGGAGGTGCACAGTGG
GGTCAGCACAGACCCGCAG
CCCCTCAAGGAGCAGCCCG
CCCTCAATGACTCCAGATA
CTGCCTGAGCAGCCGCCTG
AGGGTCTCGGCCACCTTCT
GGCAGAACCCCCGCAACCA
CTTCCGCTGTCAAGTCCAG
TTCTACGGGCTCTCGGAGA
ATGACGAGTGGACCCAGGA
TAGGGCCAAACCCGTCACC
CAGATCGTCAGCGCCGAGG
CCTGGGGTAGAGCAGACTG
TGGCTTTACCTCGGTGTCC
TACCAGCAAGGGGTCCTGT
CTGCCACCATCCTCTATGA
GATCCTGCTAGGGAAGGCC
ACCCTGTATGCTGTGCTGG
TCAGCGCCCTTGTGTTGAT
GGCCATGGTCAAGAGAAAG
GATTTCTGA
(SEQ ID NO:
TCACAACTCTCCCCAGAGAA
GGTGGTGTGAGGCCATCACG
GAVVSQHPSRV
GAAGATGCTGCTGCTTCTGC
ICKSGTSVKIEC
TGCTTCTGGGGCCAGGCTCC
RSLDFQATTMF
GGGCTTGGTGCTGTCGTCTCT
WYRQFPKQSL
MLMATSNEGS
KATYEQGVEK
DKFLINHASLTL
STLTVTSAHPE
DSSFYICSARIG
QGFMNEQFFGP
GTRLTVLEDLK
NVFPPEVAVFE
PSEAEISHTQK
ATLVCLATGF
YPDHVELSWW
VNGKEVHSGV
STDPQPLKEQP
ALNDSRYCLSS
RLRVSATFWQ
NPRNHFRCQV
QFYGLSENDE
ACCTGAAAAACGTGTTCCC
WTQDRAKPVT
ACCCGAGGTCGCTGTGTTT
QIVSAEAWGR
GAGCCATCAGAAGCAGAGA
ADCGFTSESYQ
TCTCCCACACCCAAAAGGC
QGVLSATILYE
CACACTGGTGTGCCTGGCC
ILLGKATLYAV
ACAGGCTTCTACCCCGACC
LVSALVLMAM
ACGTGGAGCTGAGCTGGTG
VKRKDSRG
GGTGAATGGGAAGGAGGT
GCACAGTGGGGTCAGCACA
GACCCGCAGCCCCTCAAGG
AGCAGCCCGCCCTCAATGA
CTCCAGATACTGCCTGAGC
AGCCGCCTGAGGGTCTCGG
CCACCTTCTGGCAGAACCC
CCGCAACCACTTCCGCTGT
CAAGTCCAGTTCTACGGGC
TCTCGGAGAATGACGAGTG
GACCCAGGATAGGGCCAAA
CCTGTCACCCAGATCGTCA
GCGCCGAGGCCTGGGGTA
GAGCAGACTGTGGCTTCAC
CTCCGAGTCTTACCAGCAA
GGGGTCCTGTCTGCCACCA
TCCTCTATGAGATCTTGCT
AGGGAAGGCCACCTTGTAT
GCCGTGCTGGTCAGTGCCC
TCGTGCTGATGGCCATGGT
CAAGAGAAAGGATTCCAGA
GGCTAG
(SEQ ID NO: 435)
GAVVSQHPSRV
ICKSGTSVKIEC
ACTCTCCCCAGAGAAGGTGG
RSLDFQATTMF
TGTGAGGCCATCACGGAAGA
WYRQFPKKSL
TGCTGCTGCTTCTGCTGCTTC
MLMATSNEGS
TGGGGCCAGGCTCCGGGCTT
KATYEQGVEK
DKFLINHASLTL
STLTVTSAHPE
DSSFYICSAPG
WRGTEAFFGQG
TRLTVVEDLNK
VFPPEVAVFEP
SEAEISHTQKA
TLVCLATGFFP
DHVELSWWVN
GKEVHSGVST
DPQPLKEQPA
LNDSRYCLSSR
LRVSATFWQN
PRNHFRCQVQ
FYGLSENDEW
TQDRAKPVTQI
VSAEAWGRAD
CGFTSVSYQQ
GTGTTCCCACCCGAGGTCG
GVLSATILYEI
CTGTGTTTGAGCCATCAGA
LLGKATLYAV
AGCAGAGATCTCCCACACC
LVSALVLMAM
CAAAAGGCCACACTGGTGT
VKRKDF
GCCTGGCCACAGGCTTCTT
CCCTGACCACGTGGAGCTG
AGCTGGTGGGTGAATGGG
AAGGAGGTGCACAGTGGG
GTCAGCACAGACCCGCAGC
CCCTCAAGGAGCAGCCCGC
CCTCAATGACTCCAGATAC
TGCCTGAGCAGCCGCCTGA
GGGTCTCGGCCACCTTCTG
GCAGAACCCCCGCAACCAC
TTCCGCTGTCAAGTCCAGT
TCTACGGGCTCTCGGAGAA
TGACGAGTGGACCCAGGAT
AGGGCCAAACCCGTCACCC
AGATCGTCAGCGCCGAGGC
CTGGGGTAGAGCAGACTGT
GGCTTTACCTCGGTGTCCT
ACCAGCAAGGGGTCCTGTC
TGCCACCATCCTCTATGAG
ATCCTGCTAGGGAAGGCCA
CCCTGTATGCTGTGCTGGT
CAGCGCCCTTGTGTTGATG
GCCATGGTCAAGAGAAAGG
ATTTCTGA
(SEQ ID NO: 442)
VSQTPKYLVTQ
MGNDKSIKCEQ
NLGHDTMYWY
KQDSKKFLKIM
CTCCTCTGCTGTGTGGTCTTC
FSYNNKELIINE
TGCCTCCTCCAAGCAGGTCC
TVPNRFSPKSPD
CTTGGACACAGCTGTTTCCC
KAHLNLHINSL
ELGPSAVYFCA
SSQPGLEQYFG
PGTRLTVTEDL
KNVFPPKVAVK
EPSEAEISHTQ
KATLVCLATG
FYPPHVELSW
WVNGKEVHSG
VSTPPQPLKEQ
PALNPSRVCLS
SRLRVSATFW
QNPRNHFRCQ
VQFVGLSENP
EWTQDRAKPV
TQIVSAEAWG
RADCGFTSESY
GACCTGAAAAACGTGTTCC
QQGVLSATILY
CACCCGAGGTCGCTGTGTT
EILLGKATLYA
TGAGCCATCAGAAGCAGAG
VLVSALVLMA
ATCTCCCACACCCAAAAGG
MVKRKPSRG
CCACACTGGTATGCCTGGC
CACAGGCTTCTACCCCGAC
CACGTGGAGCTGAGCTGGT
GGGTGAATGGGAAGGAGG
TGCACAGTGGGGTCAGCAC
AGACCCGCAGCCCCTCAAG
GAGCAGCCCGCCCTCAATG
ACTCCAGATACTGCCTGAG
CAGCCGCCTGAGGGTCTCG
GCCACCTTCTGGCAGAACC
CCCGCAACCACTTCCGCTG
TCAAGTCCAGTTCTACGGG
CTCTCGGAGAATGACGAGT
GGACCCAGGATAGGGCCA
AACCTGTCACCCAGATCGT
CAGCGCCGAGGCCTGGGG
TAGAGCAGACTGTGGCTTC
ACCTCCGAGTCTTACCAGC
AAGGGGTCCTGTCTGCCAC
CATCCTCTATGAGATCTTG
CTAGGGAAGGCCACCTTGT
ATGCCGTGCTGGTCAGTGC
CCTCGTGCTGATGGCCATG
GTCAAGAGAAAGGATTCCA
GAGGCTAG
(SEQ ID NO: 449)
VSQTPKYLVTQ
MGNDKSIKCEQ
NLGHDTMYWY
KQDSKKFLKIM
AGGCTCCTCTGCTGTGTGGT
FSYNNKELIINE
CTTCTGCCTCCTCCAAGCAG
TVPNRFSPKSPD
GTCCCTTGGACACAGCTGTT
KAHLNLHINSL
ELGDSAVYFCA
SSQEGSQDHTS
GRATEQYFGPG
TRLTVTEDLKN
VFPPEVAVFEP
SEAEISHTQKA
TLVCLATGFYP
DHVELSWWVN
GKEVHSGVST
DPQPLKEQPA
LNDSRYCLSSR
LRVSATFWQN
PRNHFRCQVQ
FYGLSENDEW
TQDRAKPVTQI
VSAEAWGRAD
CGFTSESYQQ
GVLSATILYEI
AACGTGTTCCCACCCGAGG
LLGKATLYAV
TCGCTGTGTTTGAGCCATC
LVSALVLMAM
AGAAGCAGAGATCTCCCAC
VKRKDSRG
ACCCAAAAGGCCACACTGG
TGTGCCTGGCCACAGGCTT
CTACCCCGACCACGTGGAG
CTGAGCTGGTGGGTGAATG
GGAAGGAGGTGCACAGTG
GGGTCAGCACAGACCCGCA
GCCCCTCAAGGAGCAGCCC
GCCCTCAATGACTCCAGAT
ACTGCCTGAGCAGCCGCCT
GAGGGTCTCGGCCACCTTC
TGGCAGAACCCCCGCAACC
ACTTCCGCTGTCAAGTCCA
GTTCTACGGGCTCTCGGAG
AATGACGAGTGGACCCAGG
ATAGGGCCAAACCTGTCAC
CCAGATCGTCAGCGCCGAG
GCCTGGGGTAGAGCAGACT
GTGGCTTCACCTCCGAGTC
TTACCAGCAAGGGGTCCTG
TCTGCCACCATCCTCTATG
AGATCTTGCTAGGGAAGGC
CACCTTGTATGCCGTGCTG
GTCAGTGCCCTCGTGCTGA
TGGCCATGGTCAAGAGAAA
GGATTCCAGAGGCTAG
CAGAATGACTTCCTTGAGAG
TCCTGCTCCCCTTTCATCAAT
GCACAGATACAGAAGACCCC
TCCGTCATGCAGCATCTGCC
PKFQVLKTGQS
ATGAGCATCGGCCTCCTGTG
MTLQCAQDMN
CTGTGCAGCCTTGTCTCTCCT
HEYMSWYRQD
GTGGGCAGGTCCAGTGAATG
PGMGLRLIHYS
VGAGITDQGEV
PNGYNVSRSTT
EDFPLRLLSAAP
SQTSVYFCASSL
TEGRSEQYFGP
GTRLTVTEDLK
NVFPPEVAVFE
PSEAEISHTQK
ATLVCLATGF
YPDHVELSWW
VNGKEVHSGV
STDPQPLKEQP
ALNDSRYCLSS
RLRVSATFWQ
NPRNHFRCQV
QFYGLSENDE
WTQDRAKPVT
CCTGAAAAACGTGTTCCCA
Q
IVSAEAWGR
CCCGAGGTCGCTGTGTTTG
ADCGFTSESYQ
AGCCATCAGAAGCAGAGAT
QGVLSATILYE
CTCCCACACCCAAAAGGCC
ILLGKATLYAV
ACACTGGTGTGCCTGGCCA
LVSALVLMAM
CAGGCTTCTACCCCGACCA
VKRKDSRG
CGTGGAGCTGAGCTGGTG
GGTGAATGGGAAGGAGGT
GCACAGTGGGGTCAGCAC
GGACCCGCAGCCCCTCAAG
GAGCAGCCCGCCCTCAATG
ACTCCAGATACTGCCTGAG
CAGCCGCCTGAGGGTCTCG
GCCACCTTCTGGCAGAACC
CCCGCAACCACTTCCGCTG
TCAAGTCCAGTTCTACGGG
CTCTCGGAGAATGACGAGT
GGACCCAGGATAGGGCCA
AACCTGTCACCCAGATCGT
CAGCGCCGAGGCCTGGGG
TAGAGCAGACTGTGGCTTC
ACCTCCGAGTCTTACCAGC
AAGGGGTCCTGTCTGCCAC
CATCCTCTATGAGATCTTG
CTAGGGAAGGCCACCTTGT
ATGCCGTGCTGGTCAGTGC
CCTCGTGCTGATGGCCATG
GTCAAGAGAAAGGATTCCA
GAGGCTAG
(SEQ ID NO: 463)
CACAACTCTCCCCAGAGAAG
GTGGTGTGAGGCCATCACGG
GAVVSQHPSRV
AAGATGCTGCTGCTTCTGCT
ICKSGTSVKIEC
GCTTCTGGGGCCAGGCTCCG
RSLDFQATTMF
GGCTTGGTGCTGTCGTCTCTC
WYRQFPKKSL
MLMATSNEGS
KATYEQGVEK
DKFLINHASLTL
STLTVTSAHPE
DSSFYICSAGGF
SGNTIYFGEGS
WLTVVEDLNK
VFPPEVAVFEP
SEAEISHTQKA
TLVCLATGFYP
DHVELSWWVN
GKEVHSGVST
DP
Q
PLKEQPA
LNDSRYCLSSR
LRVSATFWQN
PRNHFRCQVQ
FYGLSENDEW
CAAGGTGTTCCCACCCGAG
T
Q
DRAKPVTQI
GTCGCTGTGTTTGAGCCAT
VSAEAWGRAD
CAGAAGCAGAGATCTCCCA
CGFTSVSYQQ
CACCCAAAAGGCCACACTG
GVLSATILYEI
GTGTGCCTGGCCACAGGCT
LLGKATLYAV
TCTACCCCGACCACGTGGA
LVSALVLMAM
GCTGAGCTGGTGGGTGAAT
VKRKDF
GGGAAGGAGGTGCACAGT
GGGGTCAGCACAGACCCG
CAGCCCCTCAAGGAGCAGC
CCGCCCTCAATGACTCCAG
ATACTGCCTGAGCAGCCGC
CTGAGGGTCTCGGCCACCT
TCTGGCAGAACCCCCGCAA
CCACTTCCGCTGTCAAGTC
CAGTTCTACGGGCTCTCGG
AGAATGACGAGTGGACCCA
GGATAGGGCCAAACCCGTC
ACCCAGATCGTCAGCGCCG
AGGCCTGGGGTAGAGCAG
ACTGTGGCTTTACCTCGGT
GTCCTACCAGCAAGGGGTC
CTGTCTGCCACCATCCTCT
ATGAGATCCTGCTAGGGAA
GGCCACCCTGTATGCTGTG
CTGGTCAGCGCCCTTGTGT
TGATGGCCATGGTCAAGAG
AAAGGATTTCTGA
(SEQ ID
Vector Design
Unique complementary determining regions (CDR) from selected TCR α/β pairs will be combined with the associated constant regions to form a full length TCR DNA sequence. Sequences will have TCRα with the stop codon removed followed by a furin F2A cleavage site and then a TCRβ containing a stop codon. Native leader sequences will be used for each TCR sequence. The DNA will be synthesized and cloned into an EF1a promotor vector with puromycin resistance to produce lenti-virus constructs (VectorBuilder).
Lentiviral Transduction of TCR KO Jurkat Cell Lines
Day 1: TCR KO Jurkat cells will be removed from flasks and counted. The cells will be washed by centrifuging at 1500 rpm for five minutes and resuspended in R10 (R10 Media: Gibco, 61870 supplemented with 10% FBS Gibco, 16140 and 1× Sodium Pyruvate Gibco, 11360) at a density of 1.5e6 cells/mL. 500 μL of cells will be seeded (for a density of 7.5e5 cells/well) to appropriate wells of a 12 well plate(s) according to plate layout. In a BSL2+ environment, lentiviral constructs will be diluted in R10 media with 2× (final concentration: 4 μg/mL) Polybrene (Millipore, Cat #: TR-1003-G) to concentrations of 20 Multiplicity of Infection (MOI) per 500 μL. 500 μL of each construct will be added to appropriate wells according to plate layout. Plate(s) will be centrifuged at 2000 rpm for two hours at 30° C. After centrifuging 1 mL R10 will be added to each well. Plates will be incubated at 37° C./5% CO2 overnight.
Day 2: Cells will be transferred to individual 15 mL tubes and washed by centrifuging at 300×g for five minutes. Pellets will be resuspended in DPBS (Gibco, 14140). Washes will be repeated for a total of three washes. After the final wash, cells will be resuspended in 1 mL R10. Cells will be transferred to a 6 well plate and add 1 mL of prepared R10 media containing 2× (final concentration 1.25 μg/mL) puromycin (Gibco, A11138-03). Cells will be returned to the incubator at 37° C./5% CO2.
Day 5: Cells will be counted, and density will be scaled to be 2e5 cells/mL in an appropriately sized plate/flask for the volume. For healthy cells, puromycin concentration may be increased from 1.25 μg/mL to 2.5 μg/mL. Cells will be returned to the incubator at 37° C./5% CO2.
Day 8 (and every 3-4 days after): Cells will be counted and scaled to maintain a density of 2e5 cells/mL. Puromycin will be gradually increased up to a final concentration of 5 μg/mL.
A TCR engagement assay will be performed to determine if the TCRα/β pair is able to bind to a defined peptide-MHC (pMHC) complex and induce a T-cell response. Antigen presenting cells (APC) will be harvested and plated into white clear bottom 96 well plates at 3e5 cells/well in 40 μL in RPMI 1640+10% FBS). Peptides will be prepared by adding Dimethylsulfoxide (DMSO) or water to lyophilized powder to achieve a 10 mg/mL concentration. Peptides will be exogenously loaded onto APCs by diluting to a 10× final concentration in media and adding 10 μL to the wells. DMSO or water and/or wild type (wt) peptide at the same concentrations will be used as controls. The cells will be placed in a 37° C., 5% CO2 incubator for 2 hours. Meanwhile, reporter T-cells expressing a single TCR α/b pair from Table 3 will be harvested and diluted to 3e6 cells/mL. After the 2 hour incubation, 50 μL of T-cells will be added to each well, resulting in a 2:1 APC:T-cell ratio. Cells will be returned to the incubator for 5-6 hours. After the incubation, 100 μL of BioGlo luciferase reagent (Promega, cat #G7940) will be added to each well. Plates will be placed on a rotary plate shaker at medium speed for 2 minutes. Plates will be read on an Envision (Perkin Elmer) plate reader using a 0.5 sec luminescence detection. Cross-talk corrected values will be analyzed using GraphPad Prism graphing software.
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 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: 7, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 245;
(b) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 14, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 252;
(c) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 21, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 259;
(d) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 28, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 266;
(e) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 35, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 273;
(f) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 42, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 280;
(g) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 49, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 287;
(h) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 56, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 294;
(i) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 63, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 301;
(j) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 70, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 308;
(k) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 77, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 315;
(l) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 84, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 322;
(m) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 91, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 329;
(n) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 98, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 336;
(o) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 105, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 343;
(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: 350;
(q) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 119, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 357;
(r) 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: 364;
(s) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 133, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 371;
(t) 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: 378;
(u) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 147, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 385;
(v) 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: 392;
(w) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 161, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 399;
(x) 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: 406;
(y) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 175, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 413;
(z) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 182, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 420;
(aa) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 189, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 427;
(bb) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 196, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 434;
(cc) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 203, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 441;
(dd) 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: 448;
(ee) 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: 455;
(ff) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 224, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 462;
(gg) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 231, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 469; or
(hh) the alpha chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 238, and the beta chain comprises a CDR3 comprising the amino acid sequence of SEQ ID NO: 476.
Embodiment 2. A T-cell receptor (TCR) comprising an alpha chain and a beta chain, wherein:
(a) the alpha chain comprises a complementarity determining region 1 (CDR1) comprising the amino acid sequence of SEQ ID NO: 3, a complementarity determining region 2 (CDR2) comprising the amino acid sequence of SEQ ID NO: 5, and a complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO: 7, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 241, a CDR2 comprising the amino acid sequence of SEQ ID NO: 243, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 245;
(b) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 10, a CDR2 comprising the amino acid sequence of SEQ ID NO: 12, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 14, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 248, a CDR2 comprising the amino acid sequence of SEQ ID NO: 250, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 252;
(c) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 17, a CDR2 comprising the amino acid sequence of SEQ ID NO: 19, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 21, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 255, a CDR2 comprising the amino acid sequence of SEQ ID NO: 257, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 259;
(d) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 24, a CDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 28, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 262, a CDR2 comprising the amino acid sequence of SEQ ID NO: 264, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 266;
(e) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 31, a CDR2 comprising the amino acid sequence of SEQ ID NO: 33, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 35, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 269, a CDR2 comprising the amino acid sequence of SEQ ID NO: 271, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 273;
(f) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 38, a CDR2 comprising the amino acid sequence of SEQ ID NO: 40, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 42, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 276, a CDR2 comprising the amino acid sequence of SEQ ID NO: 278, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 280;
(g) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a CDR2 comprising the amino acid sequence of SEQ ID NO: 47, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 49, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 283, a CDR2 comprising the amino acid sequence of SEQ ID NO: 285, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 287;
(h) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 52, a CDR2 comprising the amino acid sequence of SEQ ID NO: 54, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 56, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 290, a CDR2 comprising the amino acid sequence of SEQ ID NO: 292, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 294;
(i) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 59, a CDR2 comprising the amino acid sequence of SEQ ID NO: 61, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 63, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 297, a CDR2 comprising the amino acid sequence of SEQ ID NO: 299, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 301;
(j) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 66, a CDR2 comprising the amino acid sequence of SEQ ID NO: 68, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 70, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 304, a CDR2 comprising the amino acid sequence of SEQ ID NO: 306, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 308;
(k) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 73, a CDR2 comprising the amino acid sequence of SEQ ID NO: 75, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 77, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 311, a CDR2 comprising the amino acid sequence of SEQ ID NO: 313, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 315;
(l) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 80, a CDR2 comprising the amino acid sequence of SEQ ID NO: 82, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 84, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 318, a CDR2 comprising the amino acid sequence of SEQ ID NO: 320, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 322;
(m) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 87, a CDR2 comprising the amino acid sequence of SEQ ID NO: 89, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 91, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 325, a CDR2 comprising the amino acid sequence of SEQ ID NO: 327, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 329;
(n) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 94, a CDR2 comprising the amino acid sequence of SEQ ID NO: 96, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 98, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 332, a CDR2 comprising the amino acid sequence of SEQ ID NO: 334, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 336;
(o) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 101, a CDR2 comprising the amino acid sequence of SEQ ID NO: 103, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 105, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 339, a CDR2 comprising the amino acid sequence of SEQ ID NO: 341, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 343;
(p) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 108, a CDR2 comprising the amino acid sequence of SEQ ID NO: 110, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 112, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 346, a CDR2 comprising the amino acid sequence of SEQ ID NO: 348, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 350;
(q) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 115, a CDR2 comprising the amino acid sequence of SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 119, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 353, a CDR2 comprising the amino acid sequence of SEQ ID NO: 355, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 357;
(r) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 122, a CDR2 comprising the amino acid sequence of SEQ ID NO: 124, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 126, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 360, a CDR2 comprising the amino acid sequence of SEQ ID NO: 362, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 364;
(s) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 129, a CDR2 comprising the amino acid sequence of SEQ ID NO: 131, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 133, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 367, a CDR2 comprising the amino acid sequence of SEQ ID NO: 369, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 371;
(t) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 136, a CDR2 comprising the amino acid sequence of SEQ ID NO: 138, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 140, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 374, a CDR2 comprising the amino acid sequence of SEQ ID NO: 376, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 378;
(u) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 143, a CDR2 comprising the amino acid sequence of SEQ ID NO: 145, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 147, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 381, a CDR2 comprising the amino acid sequence of SEQ ID NO: 383, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 385;
(v) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 150, a CDR2 comprising the amino acid sequence of SEQ ID NO: 152, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 154, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 388, a CDR2 comprising the amino acid sequence of SEQ ID NO: 390, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 392;
(w) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 157, a CDR2 comprising the amino acid sequence of SEQ ID NO: 159, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 161, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 395, a CDR2 comprising the amino acid sequence of SEQ ID NO: 397, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 399;
(x) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 164, a CDR2 comprising the amino acid sequence of SEQ ID NO: 166, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 168, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 402, a CDR2 comprising the amino acid sequence of SEQ ID NO: 404, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 406;
(y) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 171, a CDR2 comprising the amino acid sequence of SEQ ID NO: 173, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 175, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 409, a CDR2 comprising the amino acid sequence of SEQ ID NO: 411, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 413;
(z) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 178, a CDR2 comprising the amino acid sequence of SEQ ID NO: 180, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 182, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 416, a CDR2 comprising the amino acid sequence of SEQ ID NO: 418, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 420;
(aa) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 185, a CDR2 comprising the amino acid sequence of SEQ ID NO: 187, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 189, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 423, a CDR2 comprising the amino acid sequence of SEQ ID NO: 425, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 427;
(bb) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 192, a CDR2 comprising the amino acid sequence of SEQ ID NO: 194, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 196, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 430, a CDR2 comprising the amino acid sequence of SEQ ID NO: 432, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 434;
(cc) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 199, a CDR2 comprising the amino acid sequence of SEQ ID NO: 201, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 203, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 437, a CDR2 comprising the amino acid sequence of SEQ ID NO: 439, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 441;
(dd) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 206, a CDR2 comprising the amino acid sequence of SEQ ID NO: 208, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 210, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 444, a CDR2 comprising the amino acid sequence of SEQ ID NO: 446, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 448;
(ee) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 213, a CDR2 comprising the amino acid sequence of SEQ ID NO: 215, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 217, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 451, a CDR2 comprising the amino acid sequence of SEQ ID NO: 453, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 455;
(ff) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 220, a CDR2 comprising the amino acid sequence of SEQ ID NO: 222, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 224, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 458, a CDR2 comprising the amino acid sequence of SEQ ID NO: 460, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 462;
(gg) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 227, a CDR2 comprising the amino acid sequence of SEQ ID NO: 229, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 231, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 465, a CDR2 comprising the amino acid sequence of SEQ ID NO: 467, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 469; or
(hh) the alpha chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 234, a CDR2 comprising the amino acid sequence of SEQ ID NO: 236, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 238, and the beta chain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 472, a CDR2 comprising the amino acid sequence of SEQ ID NO: 474, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 476.
Embodiment 3. The TCR of embodiment 1 or 2, wherein:
(a) the alpha chain of embodiment 2(a) comprises a variable and joining (VJ) region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 479, and the beta chain of embodiment 2(a) comprises a variable, diversity and joining (VDJ) region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 581;
(b) the alpha chain of embodiment 2(b) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 482, and the beta chain of embodiment 2(b) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 584;
(c) the alpha chain of embodiment 2(c) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 485, and the beta chain of embodiment 2(c) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 587;
(d) the alpha chain of embodiment 2(d) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 488, and the beta chain of embodiment 2(d) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 590;
(e) the alpha chain of embodiment 2(e) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 491, and the beta chain of embodiment 2(e) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 593;
(f) the alpha chain of embodiment 2(f) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 494, and the beta chain of embodiment 2(f) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 596;
(g) the alpha chain of embodiment 2(g) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 497, and the beta chain of embodiment 2(g) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 599;
(h) the alpha chain of embodiment 2(h) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 500, and the beta chain of embodiment 2(h) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 602;
(i) the alpha chain of embodiment 2(i) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 503, and the beta chain of embodiment 2(i) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 605;
(j) the alpha chain of embodiment 2(j) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 506, and the beta chain of embodiment 2(j) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 608;
(k) the alpha chain of embodiment 2(k) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 509, and the beta chain of embodiment 2(k) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 611;
(l) the alpha chain of embodiment 2(l) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 512, and the beta chain of embodiment 2(l) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 614;
(m) the alpha chain of embodiment 2(m) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 515, and the beta chain of embodiment 2(m) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 617;
(n) the alpha chain of embodiment 2(n) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 518, and the beta chain of embodiment 2(n) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 620;
(o) the alpha chain of embodiment 2(o) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 521, and the beta chain of embodiment 2(o) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 623;
(p) the alpha chain of embodiment 2(p) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 524, and the beta chain of embodiment 2(p) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 626;
(q) the alpha chain of embodiment 2(q) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 527, and the beta chain of embodiment 2(q) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 629;
(r) the alpha chain of embodiment 2(r) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 530, and the beta chain of embodiment 2(r) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 632;
(s) the alpha chain of embodiment 2(s) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 533, and the beta chain of embodiment 2(s) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 635;
(t) the alpha chain of embodiment 2(t) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 536, and the beta chain of embodiment 2(t) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 638;
(u) the alpha chain of embodiment 2(u) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 539, and the beta chain of embodiment 2(u) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 641;
(v) the alpha chain of embodiment 2(v) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 542, and the beta chain of embodiment 2(v) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 644;
(w) the alpha chain of embodiment 2(w) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 545, and the beta chain of embodiment 2(w) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 647;
(x) the alpha chain of embodiment 2(x) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 548, and the beta chain of embodiment 2(x) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 650;
(y) the alpha chain of embodiment 2(y) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 551, and the beta chain of embodiment 2(y) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 653;
(z) the alpha chain of embodiment 2(z) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 554, and the beta chain of embodiment 2(z) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 656;
(aa) the alpha chain of embodiment 2(aa) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 557, and the beta chain of embodiment 2(aa) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 659;
(bb) the alpha chain of embodiment 2(bb) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 560, and the beta chain of embodiment 2(bb) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 662;
(cc) the alpha chain of embodiment 2(cc) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 563, and the beta chain of embodiment 2(cc) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 665;
(dd) the alpha chain of embodiment 2(dd) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 566, and the beta chain of embodiment 2(dd) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 668;
(ee) the alpha chain of embodiment 2(ee) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 569, and the beta chain of embodiment 2(ee) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 671;
(ff) the alpha chain of embodiment 2(ff) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 572, and the beta chain of embodiment 2(ff) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 674;
(gg) the alpha chain of embodiment 2(gg) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 575, and the beta chain of embodiment 2(gg) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 677; or
(hh) the alpha chain of embodiment 2(hh) comprises a VJ region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 578, and the beta chain of embodiment 2(hh) comprises a VJD region comprising an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 680.
Embodiment 4. The TCR of embodiment 1 or 2, wherein:
(a) the alpha chain of embodiment 2(a) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 477, and the beta chain of embodiment 2(a) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 579;
(b) the alpha chain of embodiment 2(b) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 480, and the beta chain of embodiment 2(b) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 582;
(c) the alpha chain of embodiment 2(c) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 483, and the beta chain of embodiment 2(c) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 585;
(d) the alpha chain of embodiment 2(d) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 486, and the beta chain of embodiment 2(d) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 588;
(e) the alpha chain of embodiment 2(e) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 489, and the beta chain of embodiment 2(e) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 591;
(f) the alpha chain of embodiment 2(f) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 492, and the beta chain of embodiment 2(f) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 594;
(g) the alpha chain of embodiment 2(g) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 495, and the beta chain of embodiment 2(g) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 597;
(h) the alpha chain of embodiment 2(h) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 498, and the beta chain of embodiment 2(h) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 600;
(i) the alpha chain of embodiment 2(i) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 501, and the beta chain of embodiment 2(i) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 603;
(j) the alpha chain of embodiment 2(j) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 504, and the beta chain of embodiment 2(j) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 606;
(k) the alpha chain of embodiment 2(k) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 507, and the beta chain of embodiment 2(k) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 609;
(l) the alpha chain of embodiment 2(l) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 510, and the beta chain of embodiment 2(l) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 612;
(m) the alpha chain of embodiment 2(m) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 513, and the beta chain of embodiment 2(m) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 615;
(n) the alpha chain of embodiment 2(n) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 516, and the beta chain of embodiment 2(n) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 618;
(o) the alpha chain of embodiment 2(o) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 519, and the beta chain of embodiment 2(o) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 621;
(p) the alpha chain of embodiment 2(p) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 522, and the beta chain of embodiment 2(p) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 624;
(q) the alpha chain of embodiment 2(q) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 525, and the beta chain of embodiment 2(q) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 627;
(r) the alpha chain of embodiment 2(r) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 528, and the beta chain of embodiment 2(r) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 630;
(s) the alpha chain of embodiment 2(s) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 531, and the beta chain of embodiment 2(s) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 633;
(t) the alpha chain of embodiment 2(t) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 534, and the beta chain of embodiment 2(t) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 636;
(u) the alpha chain of embodiment 2(u) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 537, and the beta chain of embodiment 2(u) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 639;
(v) the alpha chain of embodiment 2(v) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 540, and the beta chain of embodiment 2(v) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 642;
(w) the alpha chain of embodiment 2(w) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 543, and the beta chain of embodiment 2(w) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 645;
(x) the alpha chain of embodiment 2(x) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 546, and the beta chain of embodiment 2(x) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 648;
(y) the alpha chain of embodiment 2(y) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 549, and the beta chain of embodiment 2(y) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 651;
(z) the alpha chain of embodiment 2(z) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 552, and the beta chain of embodiment 2(z) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 654;
(aa) the alpha chain of embodiment 2(aa) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 555, and the beta chain of embodiment 2(aa) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 657;
(bb) the alpha chain of embodiment 2(bb) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 558, and the beta chain of embodiment 2(bb) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 660;
(cc) the alpha chain of embodiment 2(cc) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 561, and the beta chain of embodiment 2(cc) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 663;
(dd) the alpha chain of embodiment 2(dd) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 564, and the beta chain of embodiment 2(dd) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 666;
(ee) the alpha chain of embodiment 2(ee) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 567, and the beta chain of embodiment 2(ee) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 669;
(ff) the alpha chain of embodiment 2(ff) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 570, and the beta chain of embodiment 2(ff) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 672;
(gg) the alpha chain of embodiment 2(gg) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 573, and the beta chain of embodiment 2(gg) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 675; or
(hh) the alpha chain of embodiment 2(hh) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 576, and the beta chain of embodiment 2(hh) comprises an amino acid sequence that is at least 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to amino acid sequence of SEQ ID NO: 678.
Embodiment 5. A nucleic acid molecule encoding the TCR of any one of embodiments 1-4.
Embodiment 6. A vector comprising the nucleic acid molecule of embodiment 5.
Embodiment 7. A cell transformed to express the nucleic acid molecule of embodiment 5.
Embodiment 8. A cell comprising the vector of embodiment 6.
Embodiment 9. The cell of embodiment 7 or 8, wherein the cell is a CD8+ T cell.
Embodiment 10. A pharmaceutical composition comprising the TCR of any one of embodiment 1-4, the nucleic acid molecule of embodiment 5, the vector of embodiment 6, or the cell of any one of embodiments 7-9.
This application claims priority to U.S. Provisional Application No. 63/170,711 filed on Apr. 5, 2021 titled “CALR AND JAK2 T-CELL RECEPTORS” which is incorporated herein by reference in its entirety.
| Number | Date | Country | |
|---|---|---|---|
| 63170711 | Apr 2021 | US |
