T CELL BINDING COMPOSITIONS AND METHODS

Abstract

The disclosure generally relates to binding proteins that comprise antigen binding sites, a T cell receptor binding site, and a T cell co-stimulatory molecule binding site. The disclosure also relates to pharmaceutical compositions comprising such binding proteins, nucleic acid molecules encoding such binding proteins, and vectors comprising such nucleic acid molecules. The disclosure further relates to methods of treating a disorder or condition using such binding proteins and pharmaceutical compositions, binding proteins and pharmaceutical compositions for use in the treatment of a disorder or condition, and the use of such binding proteins and pharmaceutical compositions for the manufacture of a medicament for treating a disorder or condition.

Description

SEQUENCE LISTING STATEMENT

A computer readable form of the Sequence Listing is filed with this application by electronic submission and is incorporated into this application by reference in its entirety. The Sequence Listing is contained in the file created on Sep. 22, 2024 having the file name “23-1331-WO.xml” and is 276,466 bytes in size.

FIELD OF THE DISCLOSURE

The disclosure generally relates to binding proteins that comprise antigen binding sites, a T cell receptor binding site, and a T cell co-stimulatory molecule binding site. The disclosure also relates to pharmaceutical compositions comprising such binding proteins, nucleic acid molecules encoding such binding proteins, and vectors comprising such nucleic acid molecules. The disclosure further relates to methods of treating a disorder or condition using such binding proteins and pharmaceutical compositions, binding proteins and pharmaceutical compositions for use in the treatment of a disorder or condition, and the use of such binding proteins and pharmaceutical compositions for the manufacture of a medicament for treating a disorder or condition.

BACKGROUND OF THE DISCLOSURE

Recruitment of T cell cytotoxic activity to destroy tumor cells is a worthwhile but complicated treatment strategy for cancer. The development of CD3-based bispecific T cell engagers (TCEs) as cancer therapeutics has been ongoing for the past 30 years. TCEs simultaneously bind a tumor associated antigen (TAA) and cluster of differentiation 3 (CD3) on a T cell to form a T cell receptor (TCR)-independent artificial immune synapse, circumventing human leukocyte antigen (HLA) restriction, and inducing T cell activation and cytolysis of the tumor cell.

The first generation of TCEs were simple bispecific T cell engagers (BiTEs), composed of two tandem single-chain variable fragments (scFvs) including a strong CD3-binding arm and a TAA binding domain. To date, there is only one Food and Drug Administration-approved BiTE, blinatumomab, which targets CD3 (using the Orthoclone OKT3 antibody) and cluster of differentiation 19 (CD19). The strong in vitro cytolytic activity observed during the development of BiTEs created excitement around their potential use for treating cancer. However, the unanticipated high cytokine release syndrome (CRS) observed in the clinic somewhat tempered that excitement. Teachey et al., “Cytokine release syndrome after blinatumomab treatment related to abnormal macrophage activation and ameliorated with cytokine-directed therapy,” Blood 121: 5154-57 (2013). Another observed disadvantage of BiTE formats is that they exhibit very short half-life and have poor manufacturability. Ellerman, “Bispecific T cell engagers: Towards understanding variables influencing the in vitro potency and tumor selectivity and their modulation to enhance their efficacy and safety,” Methods 154: 102-17 (2019).

The second generation of TCEs include a fragment crystallizable (Fc) domain which can be modified to confer half-life extension and mutations to eliminate Fc receptor (FcR) binding, and present improved manufacturability. Vafa et al., “Perspective: Designing T cell Engagers With Better Therapeutic Windows,” Front. Oncol. 10: 446 (2020). Nevertheless, those molecules still include high affinity-CD3 binding domains, linking to induction of neurotoxicity and CRS in the clinic. More recent efforts have been focused on developing CD3 binding domains with reduced affinity with the hope to maintain potent T cell activation while significantly reducing associated cytokine release. Trinklein et al., “Efficient tumor killing and minimal cytokine release with novel T cell agonist bispecific antibodies,” MAbs 11: 639-52 (2019).

Importantly, both CD3-based BiTEs and novel immunoglobulin G (IgG)-format TCEs bind and activate both cluster of differentiation 4 (CD4) and cluster of differentiation 8 (CD8) T cells, potentially engaging unfavorable T cells such as regulatory T cells (Treg), which have been shown to potentially decrease the cytolytic activity of CD8 T cells. Duell et al., “Frequency of regulatory T cells determines the outcome of the T cell-engaging antibody blinatumomab in patients with B-precursor ALL,” Leukemia 31: 2181-90 (2017).

While T cell engager molecules offer promise, the therapeutic approach has faced challenges to date. There is a need in the art for improved T-cell binding proteins with increased activity and reduced off-target effects.

SUMMARY OF THE DISCLOSURE

In a first aspect the disclosure provides a binding protein comprising four polypeptide chains that form two tumor-associated antigen (TAA) binding sites, a T cell receptor binding site, and a T cell co-stimulatory molecule binding site, wherein the first and second polypeptide chains have a structure represented by the formula: V_L-C_L and a third polypeptide chain has a structure represented by the formula: V_H1-C_H1-V_HHa-F_Ca and a fourth polypeptide chain has a structure represented by the formula: V_H1-C_H1-V_HHb-F_Cb, wherein the first polypeptide and the third polypeptide form the first of the two TAA binding sites, and the second polypeptide and the fourth polypeptide form the second of the two TAA binding sites, and wherein: V_L is an immunoglobulin light chain variable domain and V_H1 is an immunoglobulin heavy chain variable domain that together form a TAA binding domain that specifically binds a tumor-associated antigen; C_L is an immunoglobulin light chain constant domain; C_H1 is an immunoglobulin CH1 heavy chain constant domain; V_HHa is a single chain variable domain that specifically binds a T cell receptor; V_HHb is a single chain variable domain that specifically binds T cell co-stimulatory molecule; F_Ca is C_H2a and C_H3a immunoglobulin heavy chain constant domains; and F_Cb is C_H2b and C_H3b immunoglobulin heavy chain constant domains.

In a second aspect the disclosure provides novel binding proteins.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings are included to provide a further understanding of the methods and compositions of the disclosure and are incorporated in and constitute a part of this disclosure. The drawings illustrate one or more aspects of the disclosure and together with the description serve to explain the principles and operation of the disclosure.

FIG. 1: FIG. 1A shows the in vitro cytolytic activity of LM1486 and CD3 engagers with monovalent or bivalent engagement to CD20. FIG. 1B shows the corresponding EC₅₀s to cytolysis in vitro assay. FIG. 1C shows corresponding CD4T cell activation profiles as % of cells expressing CD25. FIG. 1D shows corresponding CD8T cell activation profiles as % of cells expressing CD25.

FIG. 2 shows cytokine release profiles for LM1486 for IL-6 (FIG. 2A), IL-10 (FIG. 2B), TNF-α (FIG. 2C), and IL-17A (FIG. 2D).

FIG. 3 shows the in vitro cytolytic activity of LM1486 with different CD20+ B cell lines incubated with PBMCs at an E:T ratio of 5:1.

FIG. 4 illustrates the structure of LM1486 (FIG. 4A) and the structure of modified binding proteins with a null binding domain replacing the anti-CD8 domain (FIG. 4B), or a null binding domain replacing the anti-TCR binding domain (FIG. 4C). FIG. 4D shows the in vitro cytolytic activity of LM1486 and control binding proteins TENG0501 and TENG0502. FIG. 4E shows corresponding T cell activation profiles as % CD4 and CD8 T cells expressing CD25.

FIG. 5 illustrates the structure of LM1486 (FIG. 5A) and the structure of modified binding proteins with a null binding domain replacing the anti-CD20 domain on the anti-CD8 arm (FIG. 5B) or with a null binding domain replacing the anti-CD20 binding domain on the anti-TCR arm (FIG. 5C). FIG. 5D shows the in vitro cytolytic activity of LM1486 and control binding proteins TENG0499 and TENG0500. FIG. 5E shows corresponding T cell activation profiles as % CD4 and CD8 T cells expressing CD25.

FIG. 6 shows the tumor burden, total flux (p/s), which are presented for PBS treated groups (FIG. 6A), LM1486 at 0.8 mg/kg (FIG. 6B), LM1486 at 0.08 mg/kg (FIG. 6C), and LM1486 at 0.008 mg/kg (FIG. 6D).

FIG. 7 shows tumor growth inhibition (FIG. 7A), percentage survival (FIG. 7B), and percentage body weigh loss (FIG. 7C) as compared to conventional CD3 engagers in a fully humanized in vivo model with subcutaneous B cell tumor.

FIG. 8 shows the anti-tumor efficacy of LM1486 in a disseminated DLBCL in vivo model: Total Flux (FIG. 8A), TNF-α (FIG. 8B), IL-10 (FIG. 8C), IL-2 (FIG. 8D), IL-17A (FIG. 8E), and % weight loss (FIG. 8F).

FIG. 9 shows the in vitro cytolytic activity of LM1486 and LM1486-2 engagers. FIGS. 9A and 9D show cytotoxicity profiles in a range of T cell engager concentrations for 2 different PBMC donors. FIG. 9B, 9C, and FIG. 9E, 9F show associated CD4 and CD8 T cell activation profiles as % of CD25 surface expression for 2 donors, respectively.

FIG. 10 shows binding of LRC150016 to various cell lines; combined LRRC15pos cell lines (FIG. 10A), Saos-2 (FIG. 10B), and RPMI (FIG. 10C).

FIG. 11 shows LRC150016 engager mediates T-cell effector functions in an antigen (LRRC15) dependent manner. FIG. 11A shows in vitro cytolytic activity of LRC150016 against LRRC15pos (Saos-2 WT) and LRRC15neg (Saos-2 KO and A431-WT) target cell lines. FIG. 11B shows cytokine profile measured in the supernatant of T cell in vitro cytolysis experiment (as outlined in FIG. 11A) using a multiplexing cytokine assay.

FIG. 12 shows in vitro cytolysis and CD8-biased T-cell activation profiles induced by LRC150016, compared to a comparator CD3 TCE. FIG. 12A shows the in vitro cytolytic activity mediated by LRC150016 or Comparator CD3 TCE. FIG. 12B shows LRC150016 or Comparator CD3 TCE mediated CD8+ T cells activation, as measured by % of CD25 surface expression.

FIG. 12C shows LRC150016 or Comparator CD3 TCE mediated CD4+ T cells activation, as measured by % of CD25 surface expression.

FIG. 13 shows in vitro cytokine profile of LRC150016, compared to Comparator CD3 TCE: TNFα (FIG. 13A), IL-6 (FIG. 13B), IL-10 (FIG. 13C), and IL-2 (FIG. 13D).

FIG. 14 shows the LRC150016 specific in vitro cytolytic activity across an array of cell lines expressing various levels of LRRC15. FIG. 14A shows E_max and FIG. 14B shows EC₅₀.

FIG. 15 shows a diagram of the LRC150016 Binding Protein.

FIG. 16 shows in vitro cytolysis induced by LM1653 compared to a conventional GPC3×CD3 engager, against hepatocellular carcinoma (HCC) cell lines expressing different levels of GPC3. FIG. 16A shows expression of GPC3 across 4 HCC cell lines. FIG. 16B-E show the in vitro cytolytic activity of LM1653 and of a monovalent GPC3×CD3 engager against the HCC cell lines HepG2 (FIG. 16B), Hep3B (FIG. 16C), Huh-7 (FIG. 16D), and PLC/PRF/5 (FIG. 16E).

FIG. 17 shows CD8+ T cell (upper panels) and CD4+ T cell (bottom panels) activation profiles induced by LM1653 and by a monovalent GPC3×CD3 engager in cytotoxicity assay against HCC cell lines HepG2 (FIG. 17A), Hep3B (FIG. 17B), Huh-7 (FIG. 17C), and PLC/PRF/5 (FIG. 17D).

FIG. 18 shows in vitro cytokine release profiles of LM1653 compared to a conventional GPC3×CD3 engager: TNF-α (FIG. 18A), IL-10 (FIG. 18B), IL-2 (FIG. 18C), and IFN-7 (FIG. 18D), in an in vitro cytolysis assay, using HCC Hep3B cell line as target cells at E:T ratio of 10:1.

FIG. 19 shows non-specific activity of LM1653 and of a conventional GPC3×CD3 engager in a plate-bound assay. CD8 and CD4 T cell activation profiles (defined as % of parental cells expressing CD25) are depicted in FIG. 19A and FIG. 19B, respectively. Histograms represent mean±SEM; *, p<0.05, 2-way ANOVA. Released IL-6 (FIG. 19C), TNF-α (FIG. 19D), IFN-7 (FIG. 19E), IL-2 (FIG. 19F), IL-10 (FIG. 19G), and IL-17 (FIG. 19H) were measured in the supernatant at 48 hours, using a multiplex assay (Luminex). Results from one representative donor PBMC are shown.

FIG. 20 shows the in vitro cytolytic activity (FIG. 20A-B) and cytokine secretion (FIG. 20C-J) of LM1653 and of a monovalent GPC3×CD3 engager using purified CD4 T cells (upper panels) and purified CD8 T cells (bottom panels), in a Xcelligence assay.

FIG. 21 shows LM1653-dependent killing of GPC3⁺Hep3B cell line by patients-derived tumor infiltrating lymphocytes obtained from HCC stage II (FIG. 21A) and stage IIIB (FIG. 21B).

FIG. 22 shows the in vitro cytolytic activity of LM1653 and of a monovalent GPC3×CD3 engager against non small cell lung cancer lines NCI-H661 (FIG. 22A) and NCI-H2172 (FIG. 22B) and ovarian cancer cell lines OV-90 (FIG. 22C) and Kuramochi (FIG. 22D).

FIG. 23 shows comparable in vitro cytolysis and T cell activation profile induced by LM1653 and LM1653-2 (same sequence as LM1653, but a different lot). FIG. 23A-B Cytolysis activity was measured at day 4, using PBMC at the ratio of 10:1 in the XCELLIGENCE system (n=1 PBMC donor). The tables show the corresponding EC₅₀ and killing E_max for GPC3+ cell line Hep3B (FIG. 23A) and Huh-7 (FIG. 23B). T cell activation profile was determined in cytotoxicity assay, using the Hep3B (FIG. 23C-D) and Huh-7 cell line (FIG. 23E-F). The percentage of CD8 (filled symbol) and CD4 (empty symbol) T cells expressing CD25 is reported.

FIG. 24 shows that LM1653 confers anti-tumor efficacy in vivo against GPC3^high and GPC3^low HCC xenograft models. FIG. 24A shows 5e06 Hep3B cells, expressing high levels of GPC3. On day 21, animals were humanized with 10e06 panT cells expanded in vitro. Tumor burden was measured overtime and reported as mean±SEM. 1, 40% tumor-free; **, p<0.01 and ***, p<0.001, 2-way Anova. FIG. 24B shows 5e06 PLC/PRF/5 cells, expressing low levels of GPC3. Tumor burden was measured overtime and reported as mean±SEM. **, p<0.0001, 2-way Anova. Dotted lines represent TCE administrations.

FIG. 25 shows that LM1653 (GPC3 TITAN) binds human hepatoma cells Hep3B at EC₅₀: 8 nM (FIG. 25A) and HepG2 at EC₅₀: 2 nM (FIG. 25B). GPC3 TITAN binds to human PBMC at EC₅₀: 6 nM (FIG. 25C).

FIG. 26 depicts comparator CD3×CD20 T cell engagers (FIGS. 26A and 26B).

FIG. 27 depicts the superior B cell depletion (FIG. 27A and FIG. 27B), CD4 T-cell sparing activity (FIG. 27C and FIG. 27D), and selective CD8+ T-cell activation (FIG. 27E and FIG. 27F) of LM1486 (CD20 TITAN) in PBMCs derived from whole blood of patients with SLE (FIG. 27A, FIG. 27C, and FIG. 27E) and myositis (FIG. 27B, FIG. 27D, and FIG. 27F).

FIG. 28 compares the cytokine release at varying concentrations of mosunetuzumab and LM1486 (CD20 TITAN) in an assay of non-specific T-cell activation of IL-17A (FIG. 28A) and TNFα (FIG. 28B).

FIG. 29 depicts the humanized mouse model of B-cell depletion.

FIG. 30 depicts depletion of B cells in a humanized mouse model after treatment with vehicle, NIP228 (control) or varying mono doses of LM1486 (CD20 TITAN) in spleen (FIG. 30A), blood (FIG. 30B), bone marrow (FIG. 30C), kidney (FIG. 30D), thymus (FIG. 30E), and lymph nodes (FIG. 30F).

FIG. 31 shows potent in vitro cytolysis by LM1953. FIG. 31A shows the in vitro cytolytic activity of LM1953 on C4-2 cells and the corresponding EC₅₀. FIG. 31B shows the in vitro cytolytic activity of LM1953 on 22Rv1 cells and the corresponding EC₅₀. A CD8-biased T cell activation profile was induced by LM1953.

FIG. 32 shows corresponding CD4 and CD8 T cell activation profiles as % of cells expressing CD25 on the cell surface in the C4-2 cytolysis assay (FIG. 32A) and in the 22Rv1 cytolysis assay (FIG. 32B).

FIG. 33 shows in vitro cytokine release profiles of LM1953 on C4-2 cells for IL-6 (FIG. 33A) and TNF-α (FIG. 33C), and on 22Rv1 cells for IL-6 (FIG. 33B) and TNF-α (FIG. 33D).

FIG. 34 shows that LM1953 confers anti-tumor efficacy in vivo against STEAP2+ prostate cancer xenograft model 22Rv1.

FIG. 35 shows that the LRRC15 variants preferentially activate CD8+ T cells, have functional activity (FIG. 35A), are cytotoxic (FIG. 35B), and IL-6 conc (FIG. 35C).

FIG. 36 shows binding of the LRRC15 TCE variants to EphB6-expressing As293 cells.

FIG. 37 shows that plasmablasts were depleted in mice three days following a single treatment of TCP-42197 (BCMA TCE) in both the bone marrow (FIG. 37A) and in the spleen (FIG. 37B). Serum human IgG was significantly lower in the TCP-42197 treatment groups compared to the isotype control treated group (FIG. 37C).

FIG. 38 shows that TPP-42197 (BCMA TCE) highly depletes plasmablasts compared to Comparator T.

FIG. 39 shows that TPP-42197 (BCMA TCE) selectively activates CD8+ T cells (FIG. 39A) over CD4+ T cells (FIG. 39B).

FIG. 40 show that the cytokine profile of TPP-42197 (BCMA TCE) is superior to that of the Comparator T with lower cytokine secretion in general and in particular those involved in CRS events IL-17A (FIG. 40A), IL-22 (FIG. 40B), IL-10 (FIG. 40C), IL-6 (FIG. 40D), TNFα (FIG. 40E), and IFNγ (FIG. 40F).

FIG. 41 shows that LM1653-2 induced significant tumor growth inhibition in Hep3B tumor bearing mice without significant associated cytokine secretion, in comparison to a conventional GPC3×CD3 engager. FIG. 41A shows the tumor burden, which was monitored twice a week by bioluminescence imaging (BLI) until Day 24, 2 days post treatment. Serum cytokine concentrations are shown for each treatment group: IFN-7 (FIG. 41B), TNFα (FIG. 41C), IL-10 (FIG. 41D), and IL-2 (FIG. 41E).

FIG. 42 shows LM1653-3 (same sequence as LM1653 but a different lot) induced significant tumor growth inhibition in NCI-H661 lung tumor bearing mice without significant associated cytokine secretion, in comparison to vehicle-treated animals. FIG. 42A shows the tumor burden, which was monitored twice a week by bioluminescence imaging (BLI). Serum cytokine concentrations are shown for each treatment group: IFNγ (FIG. 42B), TNFα (FIG. 42C), IL-10 (FIG. 42D), and IL-2 (FIG. 42E).

FIG. 43 shows administration of LM1653 in cynomolgus monkeys results in transient CD8+ T cell margination and limited increase in circulating cytokine levels. FIG. 43A shows the absolute count of cytotoxic CD8+ T cells in blood over the course of the treatment, reported as mean±SD per group. Doses: 1 mg/kg/week IV (Δ), 6 mg/kg/week IV ( ) or 6 mg/kg/week SC (0) followed by a 2-week recovery period. Animals that received vehicle only as both IV & SC (O) were used as controls. FIGS. 43B-E shows serum IL-10 (FIG. 43B), IL-6 (FIG. 43C), MCP-1 (FIG. 43D), and MIP-1b (FIG. 43E) levels overtime, during the treatment, determined using a multiple assay (MSD) and reported as mean per treatment group. Solid and dotted lines represent values obtained for male and female, respectively.

FIG. 44 shows the schematic of the cynomolgus monkeys dosage study for LM1486 (CD20 TITAN).

FIG. 45 shows the results of B-cell depletion in blood and tissues. FIG. 45A shows the results of circulating CD19+ B cells and FIG. 45B shows tissue specific CD19+ B cells in spleen, bone marrow and lymph nodes.

FIG. 46 shows absolute circulating levels of CD8+(FIG. 46A) and CD4+(FIG. 46B) T cells post first dose, and activated CD8/CD4 (CD25+) (FIG. 46C). LM1486 and CD20 TITAN are used interchangeably.

FIG. 47 shows group mean plasma cytokine profiles for IL-6 (FIG. 47A), TNFα (FIG. 47B), and IFNγ (FIG. 47C) as a time course from Day −14 to Day 16.

FIG. 48 shows the diagrams and amino acid sequences for the various T-Cell engagers exemplified herein. LM1486-2 (CD20) (FIG. 48A), LM1653 (GPC3) (FIG. 48B), TPP-42197 (BCMA) (FIG. 48C), LM1953 (STEAP2) (FIG. 48D), TPP-46956 (LRRC15) (FIG. 48E), TPP-46957 (LRRC15) (FIG. 48F), TPP-46958 (LRRC15) (FIG. 48G), TPP-46959 (LRRC15) (FIG. 48H), TPP-46960 (LRRC15) (FIG. 48I), TPP-47826 (LRRC15) (FIG. 48J), TPP-49058 (LRRC15) (FIG. 48K) and LM1486 (FIG. 48L).

DETAILED DESCRIPTION OF THE DISCLOSURE

The disclosure generally relates to binding proteins that comprise antigen binding sites, a T cell receptor binding site, and a T cell co-stimulatory molecule binding site. The disclosure also relates to pharmaceutical compositions comprising such binding proteins, nucleic acid molecules encoding such binding proteins, and vectors comprising such nucleic acid molecules. The disclosure further relates to methods of treating a disorder or condition using such binding proteins and pharmaceutical compositions, binding proteins and pharmaceutical compositions for use in the treatment of a disorder or condition, and the use of such binding proteins and pharmaceutical compositions for the manufacture of a medicament for treating a disorder or condition.

It is to be understood that the particular aspects of the disclosure are described herein are not limited to specific aspects presented and can vary. It also will be understood that the terminology used herein is for the purpose of describing particular aspects only and, unless specifically defined herein, is not intended to be limiting. Moreover, particular aspects disclosed herein can be combined with other aspects disclosed herein, as would be recognized by a skilled person, without limitation.

Unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values herein that are expressed as ranges can assume any specific value or sub-range within the stated ranges in different aspects of the disclosure, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

Throughout this disclosure, unless the context specifically indicates otherwise, the terms “comprise” and “include” and variations thereof (e.g., “comprises,” “comprising,” “includes,” and “including”) will be understood to indicate the inclusion of a stated component, feature, element, or step or group of components, features, elements or steps but not the exclusion of any other component, feature, element, or step or group of components, features, elements, or steps.

As used herein, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly indicates otherwise. Unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular.

Percentages disclosed herein can vary in amount by +10%, 20%, or 30% from values disclosed and remain within the scope of the contemplated disclosure.

As used herein, ranges and amounts can be expressed as “about” a particular value or range. The term “about” also includes the exact amount. For example, “about 5%” means “about 5%” and also “5%.” The term “about” can also refer to +10% of a given value or range of values. Therefore, about 5% also means 4.5%-5.5%, for example.

As used herein, the terms “or” and “and/or” are utilized to describe multiple components in combination or exclusive of one another. For example, “x, y, and/or z” can refer to “x” alone, “y” alone, “z” alone, “x, y, and z,” “(x and y) or z,” “x or (y and z),” or “x or y or z.”

As utilized in accordance with the present disclosure, unless otherwise indicated, all technical and scientific terms shall be understood to have the same meaning as commonly understood by one of ordinary skill in the art.

The term “binding protein,” as used herein, refers to a non-naturally occurring (or recombinant) molecule which comprises multiple polypeptide chains that form at least one antigen binding site.

A “recombinant” molecule is one that has been prepared, expressed, created, or isolated by recombinant DNA technology means.

The term “antibody,” as used herein, refers to a protein that is capable of recognizing and specifically binding to an antigen. Ordinary or conventional mammalian antibodies comprise a tetramer, which is typically composed of two identical pairs of polypeptide chains, each pair consisting of one “light” chain (typically having a molecular weight of about 25 kDa) and one “heavy” chain (typically having a molecular weight of about 50-70 kDa). The terms “heavy chain” and “light chain,” as used herein, refer to any immunoglobulin polypeptide having sufficient variable domain sequence to confer specificity for a target antigen. The amino-terminal portion of each light and heavy chain typically includes a variable domain of about 100 to 110 or more amino acids that typically is responsible for antigen recognition. The variable domain may be subjected to further protein engineering to humanize the framework regions if the antibody was derived from a non-human source. The carboxyl-terminal portion of each chain typically defines a constant domain responsible for effector function. Thus, in a naturally occurring antibody, a full-length heavy chain immunoglobulin polypeptide includes a variable domain (V_H) and three constant domains (C_H1, C_H2, and C_H3) and a hinge region between C_H1 and C_H2, wherein the V_H domain is at the amino-terminus of the polypeptide and the CH3 domain is at the carboxyl-terminus, and a full-length light chain immunoglobulin polypeptide includes a variable domain (V_L) and a constant domain (C_L), wherein the V_L domain is at the amino-terminus of the polypeptide and the C_L domain is at the carboxyl-terminus.

Within full-length light and heavy chains, the variable and constant domains typically are joined by a “J” region of about 12 or more amino acids, with the heavy chain also including a “D” region of about 10 more amino acids. The variable regions of each light/heavy chain pair typically form an antigen binding site. The variable domains of naturally occurring antibodies typically exhibit the same general structure of relatively conserved framework regions (FR) joined by three hypervariable regions, also called complementarity determining regions or CDRs. The CDRs from the two chains of each pair typically are aligned by the framework regions, which may enable binding to a specific epitope. From the amino-terminus to the carboxyl-terminus, both light and heavy chain variable domains typically comprise the domains FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.

“Antigen-binding fragment thereof” refer to at least the minimal portion of an antibody which is capable of binding to a specified antigen which the antibody targets, e.g., at least some of the complementarity determining regions (CDRs) of the variable domain of a heavy chain (V_H) and the variable domain of a light chain (V_L) in the context of a typical antibody produced by a B cell. Antibodies or antigen-binding fragments thereof can be or be derived from polyclonal, monoclonal, human, humanized, or chimeric antibodies, single chain antibodies, epitope-binding fragments, e.g., Fab, Fab′ and F(ab′)2, Fd, Fvs, single-chain Fvs (scFvs), single-chain antibodies, disulfide-linked Fvs (sdFvs), fragments comprising either a V_L or V_H domain alone or in conjunction with a portion of the opposite domain (e.g., a whole V_L domain and a partial V_H domain with one, two, or three CDRs), and fragments produced by a Fab expression library. ScFv molecules are known in the art and are described, e.g., in U.S. Pat. No. 5,892,019.

The term “native Fc,” as used herein, refers to a molecule comprising the sequence of a non-antigen binding fragment resulting from digestion of an antibody or produced by other means, whether in monomeric or multimeric form, and can contain the hinge region. The original immunoglobulin source of the native Fc is preferably of human origin and can be any of the immunoglobulins. Native Fc molecules are made up of monomeric polypeptides that can be linked into dimeric or multimeric forms by covalent (i.e., disulfide bonds) and non-covalent association. The number of intermolecular disulfide bonds between monomeric subunits of native Fc molecules ranges from 1 to 4 depending on class (e.g., IgG, IgA, and IgE) or subclass (e.g., IgG1, IgG2, IgG3 and IgG4). One example of a native Fc is a disulfide-bonded dimer resulting from papain digestion of an IgG. The term “native Fc,” as used herein, is generic to the monomeric, dimeric, and multimeric forms.

The term “Fc variant,” as used herein, refers to a molecule or sequence that is modified from a native Fc but still comprises a binding site for the salvage receptor, FcRn (neonatal Fc receptor). Exemplary Fc variants, and their interaction with the salvage receptor, are known in the art. Thus, the term “Fc variant” can comprise a molecule or sequence that is humanized from a non-human native Fc. Furthermore, a native Fc comprises regions that can be removed or mutated to produce an Fc variant to alter certain residues that provide structural features or biological activity that are not required for the binding proteins of the disclosure. Thus, the term “Fc variant” comprises a molecule or sequence that lacks one or more native Fc sites or residues, or in which one or more Fc sites or residues has been modified, that affect or are involved in: (1) disulfide bond formation, (2) incompatibility with a selected host cell, (3) N-terminal heterogeneity upon expression in a selected host cell, (4) glycosylation, (5) interaction with complement, (6) binding to an Fc receptor other than a salvage receptor, or (7) antibody-dependent cellular cytotoxicity (ADCC).

The term “Fc,” as used herein, encompasses native Fc and Fc variants as defined above. As with Fc variants and native Fc molecules, the term “Fc” includes molecules in monomeric or multimeric form, whether digested from whole antibody or produced by other means.

Binding proteins encompassed by this disclosure can be of or be derived from any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass of immunoglobulin molecule.

The terms “knob,” “hole,” and “knob in hole” as used herein refer to a previously developed system to direct the correct pairing of heavy chains from differing antibodies. A “knob” and a “hole” are engineered into the two heavy chains of differing antibodies in order to promote correct pairing. The “knob in hole” approach is an effective way to produce bispecific antibodies by driving heterodimerization with mutations in the CH3 domain of each half antibody.

The term “antigen” or “target antigen,” as used herein, refers to a molecule or a portion of a molecule that is capable of being recognized by and bound by the antigen binding portion of the binding proteins of the disclosure. The target antigen is capable of being used in an animal to produce antibodies capable of binding to an epitope of that antigen. A target antigen may have one or more epitopes. With respect to each target antigen recognized by the antigen binding portion of the binding protein, is capable of competing with an intact antibody that recognizes the target antigen.

The term “antigen binding site,” as used herein, refers to a site created on the surface of a binding protein of the disclosure where an antigen or an epitope on an antigen is bound.

The term “linker,” as used herein, refers to one or more amino acid residues inserted between domains of the binding protein of the disclosure. For example, a linker may be inserted between domains, at the sequence level. The precise location of a domain transition can be determined by locating peptide stretches that do not form secondary structural elements such as beta-sheets or alpha-helices as demonstrated by experimental data or as can be assumed by techniques of modeling or secondary structure prediction. Linkers may or may not be needed depending on the where the stop and start residues of protein fusions are chosen because often natural linkers are found between immunoglobulin domains.

As used herein, the term “polynucleotide” includes a singular nucleic acid as well as multiple nucleic acids, and refers to an isolated nucleic acid molecule or construct, e.g., messenger RNA (mRNA) or plasmid DNA (pDNA). The term “nucleic acid” includes any nucleic acid type, such as DNA or RNA.

As used herein, the term “vector” can refer to a nucleic acid molecule as introduced into a host cell, thereby producing a transformed host cell. A vector can include nucleic acid sequences that allows the vector to replicate in a host cell, such as an origin of replication. A vector can also include one or more selectable marker gene and other genetic elements known in the art. Specific types of vector envisioned here can be associated with or incorporated into viruses to facilitate cell transformation.

As used herein, the terms “treat,” “treatment,” or “treatment of” refer to reducing disease pathology, reducing or eliminating disease symptoms, promoting increased survival rates, and/or reducing discomfort. For example, treating can refer to the ability of a therapy to reduce disease symptoms, signs, or causes when administered to a subject. Treating also refers to mitigating or decreasing at least one clinical symptom and/or inhibition or delay in the progression of the condition and/or prevention or delay of the onset of a disease or illness.

The terms “administration” or “administering,” as used herein, refer to providing, contacting, and/or delivering a binding protein by any appropriate route to achieve the desired effect. Administration may include, but is not limited to, oral, sublingual, parenteral (e.g., intravenous, subcutaneous, intracutaneous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional, or intracranial injection), transdermal, topical, buccal, rectal, vaginal, nasal, ophthalmic, via inhalation, and implants.

As used herein, the terms “subject,” “individual,” or “patient,” refer to any subject, particularly a mammalian subject, for whom diagnosis, prognosis, or therapy is desired. Mammalian subjects include, for example, humans, non-human primates, dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, bears, and so on.

As used herein, the term an “effective amount” or a “therapeutically effective amount” of an administered therapeutic substance, such as a binding protein, is an amount sufficient to carry out a specifically stated or intended purpose, such as treating or treatment of cancer. An “effective amount” can be determined empirically in a routine manner in relation to the stated purpose.

The term “pharmaceutical composition,” as used herein, refer to a compound or composition capable of inducing a desired therapeutic effect when properly administered to a subject. In some aspects, the disclosure provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of binding proteins of the disclosure. The terms “pharmaceutically acceptable carrier” or “physiologically acceptable carrier,” as used herein, refer to one or more formulation materials suitable for accomplishing or enhancing the delivery of one or more binding proteins of the disclosure.

In some aspects, the binding proteins disclosed herein may be formulated with a pharmaceutically acceptable carrier, excipient, or stabilizer, as pharmaceutical compositions. In certain aspects, such pharmaceutical compositions are suitable for administration to a human or non-human animal via any one or more routes of administration using methods known in the art. The term “pharmaceutically acceptable carrier” means one or more non-toxic materials that do not interfere with the effectiveness of the biological activity of the active ingredients. Such preparations may routinely contain salts, buffering agents, preservatives, compatible carriers, and optionally other therapeutic agents. Such pharmaceutically acceptable preparations may also contain compatible solid or liquid fillers, diluents or encapsulating substances which are suitable for administration into a human. Other contemplated carriers, excipients, and/or additives, which may be utilized in the formulations described herein include, for example, flavoring agents, antimicrobial agents, sweeteners, antioxidants, antistatic agents, lipids, protein excipients such as serum albumin, gelatin, casein, salt-forming counterions such as sodium, and the like. These and additional known pharmaceutical carriers, excipients, and/or additives suitable for use in the formulations described herein are known in the art, for example, as listed in “Remington: The Science & Practice of Pharmacy,” 21st ed., Lippincott Williams & Wilkins, (2005), and in the “Physician's Desk Reference,” 60th ed., Medical Economics, Montvale, N.J. (2005). Pharmaceutically acceptable carriers can be selected that are suitable for the mode of administration, solubility, and/or stability desired or required.

In some aspects provided herein is a binding protein comprising two tumor-associated antigen (TAA) binding sites. In some aspects, the tumor associated antigen (TAA) is cluster of differentiation 20 (CD20). CD20 is a transmembrane protein involved in Ca⁺⁺ channeling, B cell activation, and proliferation. CD20 is a membrane-embedded surface molecule which plays a role in the development and differentiation of B cells into plasma cells. In some aspects, the binding protein comprises a fragment of rituximab (see, e.g., U.S. Pat. No. 5,736,137). As used herein, CD20 is used interchangeably with LM1486.

In some aspects, the tumor associated antigen (TAA) is Glypican-3 (GPC3). GPC3 is is a Heparan Sulfate Proteoglycans (HSPG) that is highly expressed in hepatocellular carcinoma, where it can attract Wnt proteins to the cell surface and promote cell proliferation. As used herein, GPC3 is used interchangeably with LM1653.

In some aspects, the tumor associated antigen (TAA) is leucine rich repeat containing 15 (LRRC15). Leucine Rich Repeat Containing 15 (LRRC15) is a 581 amino acid type I cell membrane protein belongs to Leucine-rich repeat (LRR) super family with binding activity to extracellular matrix (ECM) components. LRRC15 is involved in cell-cell/cell-ECM interactions, cell adhesion, wound healing, and osteogenic differentiation. LRRC15 is highly expressed on cancer-associated fibroblasts (CAFs) within the stroma of various epithelial solid tumours and on mesenchymal tumours such as sarcomas, glioblastoma, and melanoma. LRRC15⁺ CAF signature is shown to be associated with PDx therapy resistance across various tumour indications like metastatic urothelial carcinoma (mUCC), renal cell cancer (RCC), head and neck squamous cell carcinoma (HNSCC), and non-small cell lung cancer (NSCLC).

In some aspects, the tumor associated antigen (TAA) is B-cell maturation antigen (BCMA), also known as tumor necrosis factor receptor superfamily member 17 (TNFRSF17). BCMA is a member of the TNF-receptor superfamily, which is preferentially expressed in mature B lymphocytes and is important for B cell development, proliferation, and survival. BCMA is associated with hematological malignancies, including, but not limited to multiple myeloma, and autoimmune/inflammatory diseases, including, but not limited to, scleroderma, systemic lupus erythematosus (SLE), myositis, rheumatoid arthritis (RA), anti-neutrophil cytoplasmic autoantibody (ANCA) vasculitis, or Sjogren's syndrome.

In some aspects, the tumor associated antigen (TAA) is six-transmembrane epithelial antigen of prostate-2 (STEAP2). STEAP2 is a membrane-embedded hemoprotein with a transmembrane domain (TMD) that chelates a heme prosthetic group, or metalloreductase. STEAP2 is associated with cancer progression by driving cellular proliferation, migration, and invasion. It also influences the transcriptional profile of genes that are part of the metastatic cascade. STEAP2 is associated with cancers, including, but not limited to prostate cancer and Ewing's sarcoma.

In some aspects provided herein is a binding protein comprising one T cell receptor (TCR) binding site. The TCR comprises a heterodimer including the highly variable alpha (a) and beta (0) chains. The multicomponent complex of the TCR comprises the CD3 co-receptor, which plays a significant role in activating T cells. In one aspect there is provided a TCR binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2) and a heavy chain CDR3 (HCDR3) comprising the amino acid sequences of SEQ ID NO: 62, SEQ ID NO: 40 and SEQ ID NO: 41 respectively. In one embodiment, the TCR binding protein comprises a VHH chain according to SEQ ID NO: 43 or SEQ ID NO: 45. In another aspect, the TCR binding protein comprises a VHH chain according to SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 47 or SEQ ID NO: 48.

In some aspects provided herein is a binding protein comprising one T cell costimulatory molecule binding site. A co-stimulatory molecule comprises a co-stimulatory domain capable of potentiating or modulating the response of immune effector cells. Co-stimulatory domains can include sequences, for example, from one or more of CD3zeta (or CD3z), CD28, CD137 (4-1BB), OX-40, ICOS, CD27, GITR, CD2, IL-2Rβ and MyD88/CD40. In some aspects, the T cell costimulatory molecule is CD8. In some aspects, the T cell costimulatory molecule is CD137 (4-1BB).

In one-non-limiting embodiment, the binding protein activates T cells only when bound to a tumor associated antigen at one or both of the tumor associated antigen binding sites.

Some aspects described herein provides a binding protein comprising four polypeptide chains that form two tumor-associated antigen (TAA) binding sites, a T cell receptor binding site, and a T cell co-stimulatory molecule binding site, wherein the first and second polypeptide chains have a structure represented by the formula: V_L-C_L and a third polypeptide chain has a structure represented by the formula: V_H1-C_H1-V_HHa-F_Ca and a fourth polypeptide chain has a structure represented by the formula: V_H1-C_H1-V_HHb-F_Cb, wherein the first polypeptide and the third polypeptide form the first of the two TAA binding sites, and the second polypeptide and the fourth polypeptide form the second of the two TAA binding sites, and wherein: V_L is an immunoglobulin light chain variable domain and V_H1 is an immunoglobulin heavy chain variable domain that together form a TAA binding domain that specifically binds a tumor-associated antigen; C_L is an immunoglobulin light chain constant domain; C_H1 is an immunoglobulin CH1 heavy chain constant domain; V_HHa is a single chain variable domain that specifically binds a T cell receptor; V_HHb is a single chain variable domain that specifically binds T cell co-stimulatory molecule; F_Ca is C_H2a and C_H3a immunoglobulin heavy chain constant domains; and F_Cb is C_H2b and C_H3b immunoglobulin heavy chain constant domains.

In some aspects, the heavy chain variable domain that specifically binds a T cell co-stimulatory molecule is an immunoglobulin heavy chain variable domain. In some aspects, the heavy chain variable domain that specifically binds a T cell co-stimulatory molecule is a single domain sequence. In particular aspects, the heavy chain variable domain that specifically binds a T cell co-stimulatory molecule is a nanobody. In particular aspects, the heavy chain variable domain that specifically binds a T cell co-stimulatory molecule is a camelid. In particular aspects, the heavy chain variable domain that specifically binds a T cell co-stimulatory molecule is a single-domain variable new antigen receptor. In various non-limiting embodiments, V_HHb comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3) comprising the amino acid sequence of SEQ ID NO: 49, SEQ ID NO: 50, and SEQ ID NO: 51 respectively; or SEQ ID NO: 169, SEQ ID NO: 170 and SEQ ID NO: 171, respectively. In various other non-limiting embodiments, V_HHb comprises an amino acid sequence at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 52 or 168, or 100% identical to SEQ ID NO: 52 or 168.

In some aspects, the V_HHb is SEQ ID NO: 52 is further modified to further enhance manufacturability. In some aspects, the V_HHb is SEQ ID NO: 52 but further comprises a D30R substitution. In some aspects, the V_HHb is SEQ ID NO: 52 but further comprises a D30P substitution. In some aspects, the V_HHb is SEQ ID NO: 52 but further comprises a S75A substitution. In some aspects, the V_HHb is SEQ ID NO: 52 but further comprises a Y102I substitution. In some aspects, the V_HHb is SEQ ID NO: 52 but further comprises a S100G substitution. In some aspects, the V_HHb is SEQ ID NO: 52 but further comprises a L101A substitution. In some aspects, the V_HHb is SEQ ID NO: 52 but further comprises a Q106N substitution. In some aspects, the V_HHb is SEQ ID NO: 52 but further comprises a D31S substitution. In some aspects, the V_HHb is SEQ ID NO: 52 but further comprises a S100P substitution. In some aspects, the V_HHb is SEQ ID NO: 52 but further comprises a R52T substitution. In some aspects, the sequence comprises one or more substitutions selected from D30R, D30P, S75A, Y102I, S100G, L101A, Q106N, D31S, S100P, ORR52T. In some aspects, the V_HHB is selected from SEQ ID NOs: 81-89.

In some aspects, the heavy chain variable domain that specifically binds a T cell receptor binding site is an immunoglobulin heavy chain variable domain. In some aspects, the heavy chain variable domain that specifically binds a T cell receptor binding site is a single domain sequence. In particular aspects, the heavy chain variable domain that specifically binds a T cell receptor binding site is a nanobody. In particular aspects, the heavy chain variable domain that specifically binds a T cell receptor binding site is a camelid. In particular aspects, the heavy chain variable domain that specifically binds a T cell receptor binding site is a single-domain variable new antigen receptor. In various non-limiting embodiments, V_HHa comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3) comprising the amino acid sequence of SEQ ID NO: 39, SEQ ID NO: 40, and SEQ ID NO: 41 respectively; SEQ ID NO: 62, SEQ ID NO: 40 and SEQ ID NO: 41 respectively; or SEQ ID NO: 173, SEQ ID NO: 174, and SEQ ID NO: 175 respectively, respectively. In various other non-limiting embodiments, V_HHa comprises the amino acid sequence at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 42-48, and 172, or any one of SEQ ID NOs: 42-48, and 172.

In some aspects, the F_Ca and/or F_Cb of the binding protein is from an IgG antibody for example IgG1, IgG2, IgG3 and IgG4. In one non-limiting embodiment F_Ca and/or the F_Cb is from an IgG1 antibody.

In some aspects, C_H3a and C_H3b are both CH3 immunoglobulin heavy chain constant domains. In some aspects, C_H3a and C_H3b in each of F_Ca and F_Cb comprise modifications to facilitate heterodimerization of F_Ca and F_Cb. In various non-limiting embodiments, the modifications are substitutions to generate a knob in one of F_Ca and F_Cb and a hole in the other of F_Ca and F_Cb. In one nonlimiting embodiment the knob is a substitution to tryptophan at position 366, and the hole is a substitution of one or more of the following: i) a substitution to valine at position 407; ii) a substitution to serine at position 366; and iii) a substitution to alanine at position 368. In another non-limiting embodiment, the F_Ca or the F_Cb containing the knob further comprises a cysteine at position 354 and/or the F_Ca or the F_Cb containing the hole comprises a cysteine at position 349 and wherein the numbering is according to the Eu index.

In some aspects, C_H2a and C_H2b are both CH2 immunoglobulin heavy chain constant domains. In some aspects, C_H2a and C_H2b immunoglobulin heavy chain constant domains each comprise the following substitutions: E233P/L234V/L235A/G236del/S267K and wherein the numbering is according to the Eu index. In other aspects, either the C_H3a immunoglobulin heavy chain constant domain or C_H3b immunoglobulin heavy chain constant domain comprises a H435R and Y436F substitution. and wherein the numbering is according to the Eu index.

In various non-limiting embodiments F_Ca comprises the amino acid sequence of SEQ ID NO: 56 or SEQ ID NO: 57. In various other non-limiting embodiments F_Cb comprises the amino acid sequence of SEQ ID NO: 58.

In some aspects, the fourth polypeptide chain comprises SEQ ID NO: 61 and the third polypeptide chain comprises either SEQ ID NO: 59 or SEQ ID NO: 60.

In some aspects, the binding protein comprises a linker. The identity and sequence of amino acid residues in the linker may vary depending on the type of secondary structural element necessary to be achieved. For example, glycine, serine, and alanine are best for linkers having maximum flexibility. Some combination of glycine, proline, threonine, and serine are useful if a more rigid and extended linker is necessary. Any amino acid residue may be considered as a linker in combination with one or more other amino acid residues, which may be the same as or different as the first amino acid residue, to construct larger peptide linkers as necessary depending on the desired properties. In some aspects described herein the binding protein can further comprise L₁, a linker positioned between C_H1 and V_HHa on the third polypeptide chain and L2, a linker positioned between V_HHa and the F_Ca on the third polypeptide chain, wherein L₁ and L2 are each independently a linker or are absent. In some aspects described herein the binding protein can further comprise L3, a linker positioned between C_H1 and V_HHb on the fourth polypeptide chain, and L4, a linker positioned between V_HHb and F_Cb on the fourth polypeptide chain, wherein L3 and L4 are each independently a linker or are absent. In various non-limiting embodiments L₁, L₂, L₃, and/or L₄ comprise one or more repeats of the amino acid sequence of SEQ ID NO: 37 and/or SEQ ID NO: 38.

In some aspects described herein the binding protein comprises H₁, an immunoglobulin hinge region positioned between C_H1 and V_HHa on the third polypeptide chain, and H₂, an immunoglobulin hinge region positioned between V_HHa and the F_Ca on the third polypeptide chain, wherein H₁ and H₂ are each independently an immunoglobulin hinge region or are absent. In various non-limiting embodiments, H₁ comprises SEQ ID NO: 53, DK (SEQ ID NO: 54), SEQ ID NO: 90 or is absent, and wherein H₂ comprises SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 90 or is absent.

In some aspects described herein the binding protein comprises H₃, an immunoglobulin hinge region positioned between C_H1 and V_HHb on the fourth polypeptide chain, and H4, an immunoglobulin hinge region positioned between V_HHb and the F_Cb on the fourth polypeptide chain, wherein H₃ and H₄ are each independently an immunoglobulin hinge region or are absent. In various non-limiting embodiments, H₃ comprises SEQ ID NO: 53, DK (SEQ ID NO: 54), SEQ ID NO: 90 or is absent, and wherein H₄ comprises SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 90, or is absent. In various non-limiting embodiments, H₃ comprises SEQ ID NO: 53, DK (SEQ ID NO: 54), SEQ ID NO: 90 or is absent, and wherein H₄ comprises SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 91, or is absent.

Some aspects described herein provide a binding protein comprising four polypeptide chains that form two tumor-associated antigen binding sites, a T cell receptor binding site, and a T cell co-stimulatory molecule binding site, wherein the first and second polypeptide chains have a structure represented by the formula: V_L-C_L, a third polypeptide chain has a structure represented by the formula: V_H1-C_H1-H₁-L₁-V_HHa-H₂-L₂-F_Ca or V_H1-C_H1-H₁-L₁-V_HHa-L₂-H₂-F_Ca, and a fourth polypeptide chain has a structure represented by the formula: V_H1-C_H1-H₃-L₃-V_HHb-H₄-L₄-F_Cb or V_H1-C_H1-H₃-L₃-V_HHb-L₄-H₄-F_Cb, wherein L₁ is a linker positioned between C_H1 and V_HHa on the third polypeptide chain, and L₂ is a linker positioned between V_HHa and the F_Ca on the third polypeptide chain, wherein L₁ and L₂ are each independently a linker or are absent, and L₃ is a linker positioned between C_H1 and V_HHb on the fourth polypeptide chain, and L₄ is a linker positioned between V_HHb and F_Cb on the fourth polypeptide chain, wherein L₃ and L₄ are each independently a linker or are absent; wherein H₁ is an immunoglobulin hinge region positioned between C_H1 and VHH on the third polypeptide chain, and H₂, an immunoglobulin hinge region positioned between V_HHa and the F_Ca on the third polypeptide chain, wherein H₁ and H₂ are each independently an immunoglobulin hinge region or are absent; wherein H₃ is an immunoglobulin hinge region positioned between C_H1 and V_HHb on the fourth polypeptide chain, and H4, an immunoglobulin hinge region positioned between V_HHb and the F_Cb on the fourth polypeptide chain, wherein H₃ and H₄ are each independently an immunoglobulin hinge region or are absent.

In some aspects, the binding protein comprises a tumor-associated antigen (TAA) which is CD20, Glypican-3 (GPC3) or leucine rich repeat containing 15 (LRRC15).

In some aspects, the tumor-associated antigen (TAA) is B-cell maturation antigen (BCMA) or six-transmembrane epithelial antigen of prostate-2 (STEAP2).

In some aspects, the TAA binding domain binds to CD20 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively. In other aspects, the TAA binding domain binds to CD20 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 182, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, respectively. In various non-limiting embodiments, the TAA binding domain comprises a V_H1 domain and a V_L domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 7 and SEQ ID NO: 8, respectively; or comprises a V_H according to SEQ ID NO: 7 and a V_L according to SEQ ID NO: 8.

In some aspects, the TAA binding domain binds to GPC3 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, and SEQ ID NO: 18, respectively. In various non-limiting embodiments, the TAA binding domain comprises a V_H1 domain and a V_L domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 19 and SEQ ID NO: 20, respectively; or comprises a V_H according to SEQ ID NO: 19 and a V_L according to SEQ ID NO: 20.

In some aspects, the TAA binding domain binds to LRRC15 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO: 30, respectively. In various non-limiting embodiments, the TAA binding domain comprises a V_H1 domain and a V_L domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 31 and SEQ ID NO: 32, respectively; or comprises a V_H according to SEQ ID NO: 31 and a V_L according to SEQ ID NO: 32.

In some aspects, the TAA binding domain binds to LRRC15 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, and SEQ ID NO: 140 respectively. In various non-limiting embodiments, the TAA binding domain comprises a V_H1 domain and a V_L domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 141 and SEQ ID NO: 142, respectively; or comprises a V_H chain according to SEQ ID NO: 141 and a V_L according to SEQ ID NO: 142.

In some aspects, the TAA binding domain binds to LRRC15 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 147, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, and SEQ ID NO: 140 respectively. In various non-limiting embodiments, the TAA binding domain comprises a V_H1 domain and a V_L domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 148 and SEQ ID NO: 142, respectively; or comprises a V_H chain according to SEQ ID NO: 148 and a V_L according to SEQ ID NO: 142.

In some aspects, the TAA binding domain binds to LRRC15 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 152, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, and SEQ ID NO: 140 respectively. In various non-limiting embodiments, the TAA binding domain comprises a V_H1 domain and a V_L domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 153 and SEQ ID NO: 142, respectively; or comprises a V_H chain according to SEQ ID NO: 153 and a V_L according to SEQ ID NO: 142.

In some aspects, the TAA binding domain binds to LRRC15 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 157, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 158, and SEQ ID NO: 140 respectively. In various non-limiting embodiments, the TAA binding domain comprises a V_H1 domain and a V_L domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 159 and SEQ ID NO: 160, respectively; or comprises a V_H chain according to SEQ ID NO: 159 and a V_L according to SEQ ID NO: 160.

In some aspects, the TAA binding domain binds to LRRC15 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 157, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 165, and SEQ ID NO: 140 respectively. In various non-limiting embodiments, the TAA binding domain comprises a V_H1 domain and a V_L domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 159 and SEQ ID NO: 166, respectively; or comprises a V_H chain according to SEQ ID NO: 159 and a V_L according to SEQ ID NO: 166.

In some aspects, the TAA binding domain binds to BCMA and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, and SEQ ID NO: 116, respectively. In various non-limiting embodiments, the TAA binding domain comprises a V_H1 domain and a V_L domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 117 and SEQ ID NO: 118, respectively; or comprises a V_H according to SEQ ID NO: 117 and a V_L according to SEQ ID NO: 118.

In some aspects, the TAA binding domain binds to STEAP-2 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, and SEQ ID NO: 128, respectively. In various non-limiting embodiments, the TAA binding domain comprises a V_H1 domain and a V_L domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 129 and SEQ ID NO: 130, respectively; or comprises a V_H chain according to SEQ ID NO: 129 and a V_L according to SEQ ID NO: 130.

Some aspects described herein provide a binding protein comprising the amino acid of sequences of SEQ ID NO: 9, SEQ ID NO: 11, and SEQ ID NO: 12.

Some aspects described herein provide a binding protein comprising the amino acid of sequences of SEQ ID NO: 21, SEQ ID NO: 23, and SEQ ID NO: 24.

Some aspects described herein provide a binding protein comprising the amino acid of sequences of SEQ ID NO: 33, SEQ ID NO: 35, and SEQ ID NO: 36.

Some aspects described herein provide a binding protein comprising the amino acid of sequences of SEQ ID NO: 119, SEQ ID NO: 121, and SEQ ID NO: 122.

Some aspects described herein provide a binding protein comprising the amino acid of sequences of SEQ ID NO: 131, SEQ ID NO: 134, and SEQ ID NO: 133.

Some aspects described herein provide a binding protein comprising the amino acid of sequences of SEQ ID NO: 143, SEQ ID NO: 145, and SEQ ID NO: 146.

Some aspects described herein provide a binding protein comprising the amino acid of sequences of SEQ ID NO: 143, SEQ ID NO: 150, and SEQ ID NO: 151.

Some aspects described herein provide a binding protein comprising the amino acid of sequences of SEQ ID NO: 143, SEQ ID NO: 155, and SEQ ID NO: 156.

Some aspects described herein provide a binding protein comprising the amino acid of sequences of SEQ ID NO: 161, SEQ ID NO: 163, and SEQ ID NO: 164.

Some aspects described herein provide a binding protein comprising the amino acid of sequences of SEQ ID NO: 167, SEQ ID NO: 163, and SEQ ID NO: 164.

Some aspects described herein provide a binding protein comprising the amino acid of sequences of SEQ ID NO: 161, SEQ ID NO: 177, and SEQ ID NO: 164.

Some aspects described herein provide a binding protein comprising the amino acid of sequences of SEQ ID NO: 143, SEQ ID NO: 176, and SEQ ID NO: 156.

In some aspects the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 9, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 11, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 12.

In some aspects the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 21, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 23, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 24.

In some aspects the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 33, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 35, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 36.

In some aspects the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 119, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 121, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 122.

In some aspects the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 131, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 134, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 133.

In some aspects the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 143, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 145, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 146.

In some aspects the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 143, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 150, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 151.

In some aspects the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 143, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 155, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 156.

In some aspects the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 161, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 163, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 164.

In some aspects the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 167, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 163, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 164.

In some aspects the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 161, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 177, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 164.

In some aspects the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 143, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 176, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 156.

Some aspects described herein provide a binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO: 30 respectively. In various non-limiting embodiments, the binding protein comprises a V_H chain according to SEQ ID NO: 31 and a V_L according to SEQ ID NO: 32. In various other non-limiting embodiments, the binding protein comprises a LC chain according to SEQ ID NO: 33 and a V_H-C_H1 to SEQ ID NO: 34. In various other non-limiting embodiments, the binding protein comprises a polypeptide chain according to SEQ ID NO: 36 and/or SEQ ID NO: 35.

Some aspects described herein provide a binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, and SEQ ID NO: 140 respectively. In various non-limiting embodiments, the binding protein comprises a V_H chain according to SEQ ID NO: 141 and a V_L according to SEQ ID NO: 142. In various other non-limiting embodiments, the binding protein comprises a LC according to SEQ ID NO: 143 and a V_H-C_H1 to SEQ ID NO: 144. In various other non-limiting embodiments, the binding protein comprises a polypeptide chain according to SEQ ID NO: 146 and/or SEQ ID NO: 145.

Some aspects described herein provide a binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 147, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, and SEQ ID NO: 140 respectively. In various non-limiting embodiments, the binding protein comprises a V_H chain according to SEQ ID NO: 148 and a V_L according to SEQ ID NO: 142. In various other non-limiting embodiments, the binding protein comprises a LC according to SEQ ID NO: 143 and a V_H-C_H1 to SEQ ID NO: 149. In various other non-limiting embodiments, the binding protein comprises a polypeptide chain according to SEQ ID NO: 151 and/or SEQ ID NO: 150.

Some aspects described herein provide a binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 152, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, and SEQ ID NO: 140 respectively. In various non-limiting embodiments, the binding protein comprises a V_H chain according to SEQ ID NO: 153 and a V_L according to SEQ ID NO: 142. In various other non-limiting embodiments, the binding protein comprises a LC according to SEQ ID NO: 143 and a V_H-C_H1 to SEQ ID NO: 154. In various other non-limiting embodiments, the binding protein comprises a polypeptide chain according to SEQ ID NO: 156 and/or SEQ ID NO: 155. In various other non-limiting embodiments, the binding protein comprises a polypeptide chain according to SEQ ID NO: 156 and/or SEQ ID NO: 176.

Some aspects described herein provide a binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 157, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 158, and SEQ ID NO: 140 respectively. In various non-limiting embodiments, the binding protein comprises a V_H chain according to SEQ ID NO: 159 and a V_L according to SEQ ID NO: 160. In various other non-limiting embodiments, the binding protein comprises a LC according to SEQ ID NO: 161 and a V_H-C_H1 to SEQ ID NO: 162. In various other non-limiting embodiments, the binding protein comprises a polypeptide chain according to SEQ ID NO: 164 and/or SEQ ID NO: 163.

Some aspects described herein provide a binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 157, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 165, and SEQ ID NO: 140 respectively. In various non-limiting embodiments, the binding protein comprises a V_H chain according to SEQ ID NO: 159 and a V_L according to SEQ ID NO: 166. In various other non-limiting embodiments, the binding protein comprises a LC according to SEQ ID NO: 167 and a V_H-C_H1 to SEQ ID NO: 162. In various other non-limiting embodiments, the binding protein comprises a polypeptide chain according to SEQ ID NO: 164 and/or SEQ ID NO: 163.

In particular aspects provided herein are amino acid sequences with a conservative substitution in which there are up to 10, up to 8, up to 5, or up to 3 amino acids substituted by amino acids having analogical or similar properties, compared to the amino acid sequence of the sequences disclosed herein.

In some aspects, the light chain (V_L-C_L) associates with the V_H and C_H1 of a heavy chain to form an “antigen binding arm” and the variable domains in the antigen binding arm interact to form the “antigen binding site”. As described herein, “antigen binding site” and “antigen binding domain” are used interchangeably.

Lambda Charge Pairs

The terms “charge pair(s)” and “charge mutation(s)” are used interchangeably throughout this specification and refer to a positively charged amino acid residue and a negatively charged amino acid residue, one of which is located in the a light chain region (e.g. constant light chain region) and the other in a heavy chain region (e.g. constant heavy chain region 1 (C_H1)) of an antigen binding arm, located at positions intended to promote association of the light and heavy chains. By “lambda charge pair”, it is meant a charge pair where a positively or negatively charged amino acid residue is located in a lambda light chain. By “kappa charge pair”, it is meant a charge pair where a positively or negatively charged amino acid residue in the light chain is located in a kappa light chain.

Without wishing to be bound by theory, it is believed that the oppositely charged amino acid residues in the charge pair increase the attraction of the heavy chain to the light chain in an antigen binding arm, thereby promoting formation of the antigen binding arm with the correct heavy and light chain.

At least one of the amino acid residues of the charge pair have been engineered into the antigen binding arm (i.e. at least one amino acid residue in the pair is not a wild-type amino acid residue). In some aspects, both amino acid residues in the charge pair are engineered into the antigen binding arm (i.e. both amino acid residues in the pair are not wild-type amino acid residues).

The amino acid residues of the charge pair are typically naturally occurring. Naturally occurring positively charged amino acid residues according to the present disclosure include arginine, lysine and histidine. Naturally occurring negatively charged amino acid residues according to the present disclosure include glutamic acid, serine, threonine and aspartic acid. Although serine and threonine are often described in the art as ‘uncharged’, they have an isoelectric point below 6 and therefore are partially negatively charged at neutral pH. For the purposes of the charge pairs disclosed herein, serine and threonine are examples of negatively charged amino acid residues (together with glutamic acid and aspartic acid).

Hence, a charge pair may comprise a positively charged amino acid residue selected from arginine, lysine or histidine located at one of the positions in the charge pair and a negatively charged amino acid residue selected from aspartic acid, glutamic acid, serine or threonine located at the other position in the charge pair. For example, the charge pair may comprise any one of the following pairs of amino acid residues: arginine and aspartic acid; arginine and glutamic acid; arginine and serine; arginine and threonine; lysine and aspartic acid; lysine and glutamic acid; lysine and serine; lysine and threonine; histidine and aspartic acid; histidine and glutamic acid; histidine and serine; and histidine and threonine.

In some aspects, the positively charged amino acid residue in the charge pair is located on the light chain and the negatively charged amino acid residue in the charge pair is located on the heavy chain. In other aspects, the negatively charged amino acid residue is located on the light chain and the positively charged amino acid residue in the charge pair is located on the heavy chain.

The binding proteins described herein may comprise a lambda charge pair in one of the antigen binding arms. As exemplified herein, lambda charge pairs can be introduced at several positions to improve pairing of the correct light and heavy chains in the antigen binding arm.

In some aspects, the lambda charge pair comprises a positively or negatively charged amino acid residue at position 117, 119, 134, 136 or 178 of the constant light chain lambda region (CLλ). In some aspects, the lambda charge pair comprises a positively or negatively charged amino acid residue at position 141, 185, 128, 145, 183, 185, 173, or 187 of the C_H1. As noted elsewhere, the numbering is according to Eu numbering. Positions 117, 119, 134, 136, and 178 of the CLλ according to Eu numbering corresponds to amino acid positions 10, 12, 27, 29, and 71 of SEQ ID NOs: 98 and 99. Positions 141, 185, 128, 145, 183, 185, 173, and 187 of the C_H1 according to Eu numbering corresponds to amino acid positions 24, 68, 11, 28, 66, 68, 56, and 70 of SEQ ID NOs: 95-99.

In some aspects, lambda charge pair located at one or more of the following pairs of positions: (i) position 117 in the CLλ and position 141 in the C_H1; (ii) position 117 in the CLλ and position 185 in the CH1; (iii) position 119 in the CLλ and position 128 in the CH1; (iv) position 134 in the CLλ and position 128 in the C_H1; (v) position 134 in the CLλ and position 145 in the C_H1; (vi) position 134 in the CLλ and position 183 in the CH1; (vii) position 136 in the CLλ and position 185 in the C_H1; (viii) position 178 in the CLλ and position 173 in the CH1; and (ix) position 117 in the CLλ and position 187 in the CH1.

In some aspects, the lambda charge pair is located at charge pair is located at position 117 in the CLλ and position 141 in the CH1. For example, the lambda charge pair can be selected from the following list: arginine at position 117 of the CLλ and aspartic acid at position 141 of the CH1; arginine at position 117 of the CLλ and glutamic acid at position 141 of the CH1; arginine at position 117 of the CLλ and serine at position 141 of the C_H1; arginine at position 117 of the CLλ and threonine at position 141 of the CH1; lysine at position 117 of the CLλ and aspartic acid at position 141 of the CH1; lysine at position 117 of the CLλ and glutamic acid at position 141 of the CH1; lysine at position 117 of the CLλ and serine at position 141 of the CH1; and lysine at position 117 of the CLλ and threonine at position 141 of the CH1.

In some aspects, the lambda charge pair is located at charge pair is located at position 117 in the CLλ and position 185 in the CH1. For example, the lambda charge pair can be selected from the following list: arginine at position 117 of the CLλ and aspartic acid at position 185 of the CH1; arginine at position 117 of the CLλ and glutamic acid at position 185 of the CH1; arginine at position 117 of the CLλ and serine at position 185 of the CH1; arginine at position 117 of the CLλ and threonine at position 185 of the CH1; lysine at position 117 of the CLλ and aspartic acid at position 185 of the CH1; lysine at position 117 of the CLλ and glutamic acid at position 185 of the CH1; lysine at position 117 of the CLλ and serine at position 185 of the CH1; and lysine at position 117 of the CLλ and threonine at position 185 of the CH1.

In some aspects, the lambda charge pair is located at charge pair is located at position 119 in the CLλ and position 128 in the CH1. For example, the lambda charge pair can be selected from the following list: arginine at position 119 of the CLλ and aspartic acid at position 128 of the CH1; arginine at position 119 of the CLλ and glutamic acid at position 128 of the CH1; arginine at position 119 of the CLλ and serine at position 128 of the CH1; arginine at position 119 of the CLλ and threonine at position 128 of the CH1; lysine at position 119 of the CLλ and aspartic acid at position 128 of the CH1; lysine at position 119 of the CLλ and glutamic acid at position 128 of the CH1; lysine at position 119 of the CLλ and serine at position 128 of the CH1; and lysine at position 119 of the CLλ and threonine at position 128 of the CH1.

In some aspects, the lambda charge pair is located at charge pair is located at position 134 in the CLλ and position 128 in the CH1. For example, the lambda charge pair can be selected from the following list: arginine at position 134 of the CLλ and aspartic acid at position 128 of the CH1; arginine at position 134 of the CLλ and glutamic acid at position 128 of the CH1; arginine at position 134 of the CLλ and serine at position 128 of the CH1; arginine at position 134 of the CLλ and threonine at position 128 of the CH1; lysine at position 134 of the CLλ and aspartic acid at position 128 of the CH1; lysine at position 134 of the CLλ and glutamic acid at position 128 of the CH1; lysine at position 134 of the CLλ and serine at position 128 of the CH1; and lysine at position 134 of the CLλ and threonine at position 128 of the CH1.

In some aspects, the lambda charge pair is located at charge pair is located at position 134 in the CLλ and position 145 in the CH1. For example, the lambda charge pair can be selected from the following list: arginine at position 134 of the CLλ and aspartic acid at position 145 of the CH1; arginine at position 134 of the CLλ and glutamic acid at position 145 of the CH1; arginine at position 134 of the CLλ and serine at position 145 of the CH1; arginine at position 134 of the CLλ and threonine at position 145 of the CH1; lysine at position 134 of the CLλ and aspartic acid at position 145 of the CH1; lysine at position 134 of the CLλ and glutamic acid at position 145 of the CH1; lysine at position 134 of the CLλ and serine at position 145 of the CH1; and lysine at position 134 of the CLλ and threonine at position 145 of the CH1.

In some aspects, the lambda charge pair is located at charge pair is located at position 134 in the CLλ and position 183 in the CH1. For example, the lambda charge pair can be selected from the following list: arginine at position 134 of the CLλ and aspartic acid at position 183 of the CH1; arginine at position 134 of the CLλ and glutamic acid at position 183 of the CH1; arginine at position 134 of the CLλ and serine at position 183 of the CH1; arginine at position 134 of the CLλ and threonine at position 183 of the CH1; lysine at position 134 of the CLλ and aspartic acid at position 183 of the CH1; lysine at position 134 of the CLλ and glutamic acid at position 183 of the CH1; lysine at position 134 of the CLλ and serine at position 183 of the CH1; and lysine at position 134 of the CLλ and threonine at position 183 of the CH1.

In some aspects, the lambda charge pair is a lysine at position 134 of the CLX, and an aspartic acid or a serine at position 183 of the CH1. In the CH1 sequences provided as SEQ ID NO: 97 or SEQ ID NO: 98, EU position 183 is a serine and therefore it is not necessary to introduce a modification in the CH1 of SEQ ID NO: 100 in order to produce a charge pair with a positively charged amino acid at position 134 of the CLX.

In some aspects, the lambda charge pair is located at charge pair is located at position 136 in the CLλ and position 185 in the CH1. For example, the lambda charge pair can be selected from the following list: arginine at position 136 of the CLλ and aspartic acid at position 185 of the CH1; arginine at position 136 of the CLλ and glutamic acid at position 185 of the CH1; arginine at position 136 of the CLλ and serine at position 185 of the CH1; arginine at position 136 of the CLλ and threonine at position 185 of the CH1; lysine at position 136 of the CLλ and aspartic acid at position 185 of the CH1; lysine at position 136 of the CLλ and glutamic acid at position 185 of the CH1; lysine at position 136 of the CLλ and serine at position 185 of the CH1; and lysine at position 136 of the CLλ and threonine at position 185 of the CH1.

In some aspects, the lambda charge pair is located at charge pair is located at position 178 in the CLλ and position 173 in the CH1. For example, the lambda charge pair can be selected from the following list: arginine at position 178 of the CLλ and aspartic acid at position 173 of the CH1; arginine at position 178 of the CLλ and glutamic acid at position 173 of the CH1; arginine at position 178 of the CLλ and serine at position 173 of the CH1; arginine at position 178 of the C_L, and threonine at position 173 of the CH1; lysine at position 178 of the C_L, and aspartic acid at position 173 of the CH1; lysine at position 178 of the C_L, and glutamic acid at position 173 of the CH1; lysine at position 178 of the C_L, and serine at position 173 of the CH1; and lysine at position 178 of the CLλ and threonine at position 173 of the CH1.

In some aspects, the first antigen binding arm comprises more than one lambda charge pair. For example, the first antigen binding arm may comprise two, three, four, five, six, seven, eight or nine lambda charge pairs at positions (i) to (ix) described above.

In various aspects, the CLλ of the first polypeptide chain and/or CLλ of the second polypeptide chain each comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NOs: 98-99.

In various other aspects, CLλ comprises an amino acid sequence according to SEQ ID NO: 105, and the corresponding C_H1 comprises an amino acid sequence according to SEQ ID NO: 104. In various other aspects, C_L, comprises an amino acid sequence according to SEQ ID NO: 105, and the corresponding C_H1 comprises an amino acid sequence according to SEQ ID NO: 220.

In various other aspects, CLλ comprises an amino acid sequence according to SEQ ID NO: 107, and the corresponding C_H1 comprises an amino acid sequence according to SEQ ID NO: 106.

Engineered Disulfides

In some aspects, the binding proteins contain engineered disulfides in addition to or instead of the lambda charge pairs. By “engineered disulfides” it is meant that a native interchain disulfide bond at the C_H1-C_L interface (e.g. at 220 of the C_H1 and 212 of the C_L) of one of the antigen binding arms has been replaced by an engineered (non-native) interchain disulfide, while the other antigen binding arm contains the native interchain disulfide bond at the C_H1-C_L interface. An engineered disulfide is typically formed by engineering cysteines into the C_L of a light chain and the C_H1 of the corresponding heavy chain and replacing the cysteines that normally form the interchain disulfide. Disclosure related to the introduction of engineered disulfide into binding proteins for the purpose of promoting heterodimerization can found e.g., in U.S. Pat. No. 9,527,927 and Mazor, 2015, which are herein incorporated by reference in their entirety.

The formation of disulfide bonds between cysteine residues occurs during the folding of many proteins that enter the secretory pathway. As the polypeptide chain collapses, cysteines brought into proximity can form covalent linkages during a process catalyzed by members of the protein disulfide isomerase family. The term “disulfide link” or “disulfide linked” as used herein, refers to the single covalent bond formed from the coupling of thiol groups, especially of cysteine residues. In some aspects, the covalent linkage between two cysteines is between the two sulfur atoms of each residue. However, depending on the environment, not all protein species may have a disulfide present at all times, for example, in the event of disulfide reduction. Thus, the term “disulfide link” or “disulfide linked” (whether native or engineered), in some aspects, also refers to the presence of two cysteine residues that are capable of forming a disulfide link, irrespective of whether or not they are actually linked at that individual point in time.

In some aspects herein, “V12 DS” or “V12” as used herein refers to removal of the native interchain disulfide bond in the C_H1/CLλ interface and the subsequent introduction of the alternative disulfide bond comprising four mutations: F126C/C220V in the C_H1 domain, and mutations S122C/C212V in the lambda constant domain.

Thus, in some aspects: (i) the C_L of the first polypeptide chain and the CH1 of the third polypeptide chain, wherein the C_L of the first polypeptide chain is a CLλ and wherein the pair of engineered cysteines are located at position 122 of the CLλ of the first polypeptide chain and position 126 of the CH1 of the third polypeptide chain, and wherein the CLλ of the first polypeptide chain comprises a non-cysteine residue at position 212 and the CH1 of the third polypeptide chain comprises a non-cysteine residue at position 220, optionally wherein the non-cysteine residues are valines; or (ii) the C_L of the second polypeptide chain and the CH1 of the fourth polypeptide chain, wherein the C_L of the first polypeptide chain is a CLλ and wherein the pair of engineered cysteines are located at position 122 of the CLλ of the second polypeptide chain and position 126 of the CH1 of the fourth polypeptide chain, and wherein the CLλ of the second polypeptide chain comprises a non-cysteine residue at position 212 and the CH1 of the fourth polypeptide chain comprises a non-cysteine residue at position 220, optionally wherein the non-cysteine residues are valines.

In some aspects, the pair of cysteines are engineered into either: (i) the C_L of the first polypeptide chain and the CH1 of the third polypeptide chain, wherein the C_L of the first polypeptide chain is a CLκ and wherein the pair of engineered cysteines are located at position 121 of the CLκ of the first polypeptide chain and position 126 of the CH1 of the third polypeptide chain, and wherein the CLκ of the first polypeptide chain comprises a non-cysteine residue at position 214 and the CH1 of the third polypeptide chain comprises a non-cysteine residue at position 220, optionally wherein the non-cysteine residues are valines; or (ii) the C_L of the second polypeptide chain and the CH1 of the fourth polypeptide chain, wherein the C_L of the first polypeptide chain is a CLκ and wherein the pair of engineered cysteines are located at position 121 of the CLκ of the second polypeptide chain and position 126 of the CH1 of the fourth polypeptide chain, and wherein the CLκ of the second polypeptide chain comprises a non-cysteine residue at position 214 and the CH1 of the fourth polypeptide chain comprises a non-cysteine residue at position 220, optionally wherein the non-cysteine residues are valines.

An exemplary amino acid sequence of a CLλ comprising an engineered cysteine is provided as SEQ ID NO: 97 and an exemplary amino acid sequence of the CH1 comprising the corresponding engineered cysteine to form the engineered disulfide is provided as SEQ ID NO: 99.

In the binding proteins antibodies exemplified herein, the engineered disulfide may be present on the “first” antigen binding arm containing the lambda charge pairs and the native disulfide is present on the “second” antigen binding arm that does not contain the lambda charge pairs. However, the opposite arrangement is also specifically contemplated, i.e. where the native disulfide is present on the first antigen binding arm and the engineered disulfide is present on the second antigen binding arm.

In some aspects, the pair of cysteines are engineered into a constant light chain kappa region (CLκ) and CH1 and are located at position 121 of the CLκ and position 126 of the CH1, and wherein the same CLκ comprises a non-cysteine residue at position 214 and the same CH1 comprises a non-cysteine residue at position 220. In some aspects, the non-cysteine residues are valines.

Kappa Charge Pairs

In some aspects, the antigen binding arm comprises a constant light chain kappa region (CLκ). As described herein, techniques such as light chain affinity chromatography that utilizes affinity resins specific for either CLκ or CLλ can be used to selectively purify antibodies based on their light chain. Examples of such affinity resins include the LambdaFabSelect and KappaSelect resins available from GE Healthcare. Such methods can be used to selectively purify binding proteins containing both CLκ and CLλ and can therefore be used to improve production of bispecific antibodies in this format.

An example of an CLκ amino acid sequence is provided as SEQ ID NO: 100.

In some aspects, one of the binding arms comprises a kappa charge pair. As described above, kappa charge pairs refer to a positively charged amino acid residue and a negatively charged amino acid residue, one of which is located in the kappa light chain (e.g. CLκ) and the other in the heavy chain (e.g. CH1) of an antigen binding arm, located at positions intended to promote association of the light chain and CH1 of the second antigen binding arm.

In some aspects, the second antigen binding arm comprises a kappa charge pair located at position 133 in the CLκ and position 183 in the CH1. In some aspects, the negatively charged amino acid residue in the kappa charge pair is at position 133 of the CLκ and the positively charged amino acid residue in the kappa charge pair 183 of the CH1. In other aspects, the positively charged amino acid residue in the kappa charge pair is at position 133 of the CLκ and the negatively charged amino acid residue in the kappa charge pair 183 of the CH1. In some aspects, the negatively charged amino acid residue (e.g. at position 133 of the CLκ) is a glutamic acid, and wherein the positively charged amino acid residue (e.g. at position 183 of the CH1) is a lysine. As noted elsewhere, this numbering is according to Eu numbering.

Position 133 of the CLκ according to Eu numbering corresponds to amino acid position 26 of SEQ ID NO: 100. Position 183 of the CH1 according to Eu numbering corresponds to amino acid position 66 of SEQ ID NOs: 95 and 97.

In certain aspects, the binding protein comprises a first antigen binding arm with a lambda charge pair as described above and a second antigen binding arm with a kappa charge pair as described above, wherein the binding protein comprises engineered disulfides. For example, in one aspect, the first antigen binding arm comprises a lambda charge pair (e.g. at position 117 in the CLλ and position 141 in the CH1) and the disulfide link between the light chain and CH1 is formed between a pair of cysteines engineered into the CLλ and CH1; and the second antigen binding arm comprises a kappa charge pair (e.g. at position 133 in the CLκ and position 183 in the CH1) and the disulfide link between the light chain and CH1 is formed between a pair of native cysteines in the CLκ of the second light chain and CH1.

In another aspect, the first antigen binding arm comprises a lambda charge pair (e.g. at position 117 in the CLλ and position 141 in the CH1) and the disulfide link between the first light chain and CH1 is formed between a pair of native cysteines in CLλ of the light chain and CH1; and the second antigen binding arm comprises a kappa charge pair (e.g. at position 133 in the CLκ and position 183 in the CH1) and the disulfide link between the light chain and CH1 of the second antigen binding arm is formed between a pair of cysteines engineered in the CLκ of the light chain and CH1.

In some aspects, the CLκ of the first polypeptide chain or the second polypeptide chain comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 100.

In various aspects, the corresponding C_H1 in each of the one or more lambda charge pairs comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to SEQ ID NO: 95-97, and/or the corresponding C_H1 in the kappa charge pair comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to SEQ ID NO: 100.

Compositions and Methods of Treatment

The binding proteins of the disclosure may be prepared using domains or sequences obtained or derived from any human or non-human antibody, including, for example, human, murine, or humanized antibodies. Some aspects of the present disclosure relate to an isolated nucleic acid molecule encoding a binding protein as described herein. In various aspects the isolated nucleic acid molecules comprise a nucleotide sequence according to any one of SEQ ID NOs: 184-219. In one aspect, the isolated nucleic acid molecule comprises the nucleotide sequence according to any one of SEQ ID NOs: 184-186. In one aspect, the isolated nucleic acid molecule comprises the nucleotide sequence according to any one of SEQ ID NOs: 187-189. In one aspect, the isolated nucleic acid molecule comprises the nucleotide sequence according to any one of SEQ ID NOs: 190-192. In one aspect, the isolated nucleic acid molecule comprises the nucleotide sequence according to any one of SEQ ID NOs: 193-196. In one aspect, the isolated nucleic acid molecule comprises the nucleotide sequence according to any one of SEQ ID NOs: 196-198.

In one aspect, the isolated nucleic acid molecule comprises the nucleotide sequence according to any one of SEQ ID NOs: 199-201. In one aspect, the isolated nucleic acid molecule comprises the nucleotide sequence according to any one of SEQ ID NOs: 202-204. In one aspect, the isolated nucleic acid molecule comprises the nucleotide sequence according to any one of SEQ ID NOs: 205-207. In one aspect, the isolated nucleic acid molecule comprises the nucleotide sequence according to any one of SEQ ID NOs: 208-210. In one aspect, the isolated nucleic acid molecule comprises the nucleotide sequence according to any one of SEQ ID NOs: 211-213. In one aspect, the isolated nucleic acid molecule comprises the nucleotide sequence according to any one of SEQ ID NOs: 214-216. In one aspect, the isolated nucleic acid molecule comprises the nucleotide sequence according to any one of SEQ ID NOs: 217-219.

The isolated nucleic acid molecule may be included in a vector. In one aspect, the vector comprises the isolated nucleic acid molecule of any one of SEQ ID NOs: 184-219.

In some aspects, the methods disclosed herein relate to treating a subject for cancer by administering an effective amount of a binding protein. The disclosure also provides a therapeutically effective amount of a binding protein for use in treating cancer in a subject. The disclosure also provides use of a binding protein and/or pharmaceutical composition for the manufacture of a medicament. The disclosure also provides use of a binding protein and/or pharmaceutical composition for the manufacture of a medicament for treating cancer.

In the method of treatment of the present disclosure, the binding proteins can preferentially activate a subset of T cells in the subject. The subset of T cells can be CD8+ T cells. The CD8+ T cells can be preferentially activated as compared to CD4 T cells. The preferential activation of CD8+ T cells reduces engagement with tumor promoting T cells and CD4+ T cells which produce the majority of cytokines that cause release syndrome (CRS). Additionally, the preferential engagement of the CD8+ T cells induce pyroptosis.

The activation of T cells through methods of the present disclosure can be determined by measuring the percentage of surface interleukin-2 receptor alpha chain-positive (CD25+) T cells. The percentage of surface CD25+ T cells that are CD8 T cells can be higher than the percentage of surface CD25+ T cells that are CD4 T cells. In particular aspects, activation of T cells can be determined by the percentage of CD69+/CD25+ T cells. In particular aspects, activation of T cells can be determined by measuring levels of cytokines released by activated T cells.

The method of treatment of the present disclosure can result in reduced engagement of regulatory T cells (Tregs), increased cytolytic activity, and/or reduced incidence of cytokine release syndrome (CRS) relative to that resulting from bispecific T-cell engager (BiTEs) previously known in the art. The disclosure also provides a therapeutically effective amount of a binding protein for use in reducing engagement of regulatory T cells (Tregs), increasing cytolytic activity, and/or reducing incidence of cytokine release syndrome (CRS) relative to that resulting from bispecific T-cell engager (BiTEs) previously known in the art.

In some aspects, the cancer is a B-cell malignancy, liver cancer, hepatocellular carcinoma (HCC), lung cancer, non-small cell lung cancer (NSCLC), squamous non-small cell lung cancer (sqNSCLC), ovarian cancer, clear cell ovarian cancer, carcinoma, Merkel cell carcinoma, gastric cancer, hepatoblastoma, nephroblastoma, melanoma, sarcoma, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial cell carcinoma, osteosarcoma, glioblastoma, thyroid cancer, multiple myeloma, prostate cancer or Ewing's sarcoma.

In some aspects, the cancer includes B cell malignancies, liver cancer, hepatocellular carcinoma (HCC), melanoma, sarcoma, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial cell carcinoma, osteosarcoma, glioblastoma, lung cancer, non-small cell lung cancer, and/or thyroid cancer. In some aspects, the cancer includes B cell malignancies, liver cancer, hepatocellular carcinoma (HCC), melanoma, sarcoma, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial cell carcinoma, osteosarcoma, glioblastoma, lung cancer, non-small cell lung cancer, multiple myeloma, prostate cancer, Ewing's sarcoma, ovarian cancer and/or thyroid cancer.

In some aspects, the B cell malignancy, including chronic malignancy, is non-Hodgkin's lymphoma (NHL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Burkett lymphoma, or primary mediastinal large B cell lymphoma (PMBCL).

In some aspects, the B cell malignancy, including chronic malignancy is non-Hodgkin's lymphoma (NHL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Burkett lymphoma, primary mediastinal large B cell lymphoma (PMBCL) or small lymphocytic leukemia (SLL).

In various non-limiting embodiments, the CLL, MCL, FL, and/or DLBCL is positive for AA amyloidosis. In various other non-limiting embodiments, the CLL, MCL, FL, and/or DLBCL is fibrin-associated (FA).

In some embodiments the cancer is a B cell malignancy. In another embodiment, the cancer is hepatocellular carcinoma. In another embodiment, the cancer is multiple myeloma. In a further embodiment, the cancer is osteosarcoma.

In some aspects the tumor-associated antigen is CD20 and the cancer is a B-cell malignancy. In various non-limited embodiments the B-cell malignancy is non-Hodgkin's lymphoma (NHL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Burkett lymphoma, or primary mediastinal large B cell lymphoma (PMBCL). In another embodiment the B-cell malignancy is small lymphocytic leukemia (SLL).

In some aspects, the tumor-associated antigen is Glypican-3 (GPC3) and the cancer is liver cancer or hepatocellular carcinoma (HCC), non-small cell lung cancer (NSCLC), squamous non-small cell lung cancer (sqNSCLC), ovarian cancer, clear cell ovarian cancer, carcinoma, Merkel cell carcinoma, gastric cancer, hepatoblastoma, or nephroblastoma.

In some aspects, the tumor-associated antigen is leucine rich repeat containing 15 (LRRC15) and the cancer is melanoma, sarcoma, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial cell carcinoma, osteosarcoma, glioblastoma, lung cancer, non-small cell lung cancer, or thyroid cancer.

In some aspects, the tumor-associated antigen is B cell maturation antigen (BCMA) and the cancer is multiple myeloma.

In some aspects, the tumor-associated antigen is STEAP2 and the cancer is prostate cancer or Ewing's sarcoma.

In some aspects, the methods disclosed herein relate to treating an inflammatory disease in a subject in need thereof by administering an effective amount of a binding protein. The disclosure also provides a therapeutically effective amount of a binding protein for use in treating an inflammatory disease in a subject. The disclosure also provides use of a binding protein and/or pharmaceutical composition for the manufacture of a medicament. The disclosure also provides use of a binding protein and/or pharmaceutical composition for the manufacture of a medicament for treating an inflammatory disease.

In some aspects, the methods disclosed herein relate to treating an inflammatory disease and/or autoimmune disorder in a subject in need thereof by administering an effective amount of a binding protein. The disclosure also provides a therapeutically effective amount of a binding protein for use in treating an inflammatory disease and/or autoimmune disorder in a subject. The disclosure also provides use of a binding protein and/or pharmaceutical composition for the manufacture of a medicament. The disclosure also provides use of a binding protein and/or pharmaceutical composition for the manufacture of a medicament for treating an inflammatory disease and/or autoimmune disorder.

In some aspects, the inflammatory disease and/or autoimmune disorder is, but is not limited to, systemic lupus erythematosus (SLE), myositis, rheumatoid arthritis (RA), Sjogren's syndrome, or anti-neutrophil cytoplasmic autoantibody (ANCA) vasculitis. In one embodiment the inflammatory disease and/or autoimmune disorder is systemic lupus erythematosus (SLE). In another embodiment the inflammatory disease and/or autoimmune disorder is myositis. In another embodiment the inflammatory disease and/or autoimmune disorder is rheumatoid arthritis (RA). In another embodiment the inflammatory disease and/or autoimmune disorder is Sjogren's syndrome. In a further embodiment the inflammatory disease and/or autoimmune disorder is anti-neutrophil cytoplasmic autoantibody (ANCA) vasculitis.

In some aspects, the methods disclosed herein relate to treating a subject for an inflammatory disease and/or autoimmune disorder where B cells are involved by administering an effective amount of a binding protein. The disclosure also provides a therapeutically effective amount of a binding protein for use in treating an inflammatory disease where B cells are involved in a subject.

In some aspects, the tumor-associated antigen is CD20 and the inflammatory disease or autoimmune disorder involves B cells.

In some aspects, the tumor-associated antigen is B cell maturation antigen (BCMA) and the autoimmune disorder is scleroderma, systemic lupus erythematosus (SLE), myositis, rheumatoid arthritis (RA) or Sjogren's syndrome.

In various aspects the binding proteins, compositions, pharmaceutical compositions, and/or medicaments can be administered using any suitable route of administration, including but not limited to oral, sublingual, parenteral (e.g., intravenous, subcutaneous, intracutaneous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional, or intracranial injection), transdermal, topical, buccal, rectal, vaginal, nasal, ophthalmic, via inhalation, and implants.

Claims or descriptions that include “or” between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context. The disclosure includes aspects in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The disclosure includes aspects in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process.

Furthermore, the disclosure encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims is introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any elements can be removed from the group.

It should it be understood that, in general, where the disclosure, or aspects of the disclosure, is/are referred to as comprising particular elements and/or features, certain aspects of the disclosure or aspects of the disclosure consist, or consist essentially of, such elements and/or features. For purposes of simplicity, those aspects have not been specifically set forth in haec verba herein.

In view of the present disclosure, the binding proteins, pharmaceutical compositions, nucleic acid molecules, vectors, methods, compositions for use, and medicaments described herein can be configured by the person of ordinary skill in the art to meet the desired need.

Without limiting the disclosure, a number of embodiments of the disclosure are described herein for purpose of illustration.

Embodiment 1. A binding protein comprising four polypeptide chains that form two tumor-associated antigen (TAA) binding sites, a T cell receptor binding site, and a T cell co-stimulatory molecule binding site, wherein the first and second polypeptide chains have a structure represented by the formula: V_L-C_L a third polypeptide chain has a structure represented by the formula: V_H1-C_H1-V_HHa-F_Ca and a fourth polypeptide chain has a structure represented by the formula: V_H1-C_H1-V_HHb-F_Cb wherein the first polypeptide and the third polypeptide form the first of the two TAA binding sites, and the second polypeptide and the fourth polypeptide form the second of the two TAA binding sites, and wherein: V_L is an immunoglobulin light chain variable domain and V_H1 is an immunoglobulin heavy chain variable domain that together form a TAA binding domain that specifically binds a tumor-associated antigen; C_L is an immunoglobulin light chain constant domain; C_H1 is an immunoglobulin CH1 heavy chain constant domain; V_HHa is a single chain variable domain that specifically binds a T cell receptor; V_HHb is a single chain variable domain that specifically binds T cell co-stimulatory molecule; F_Ca is C_H2a and C_H3a immunoglobulin heavy chain constant domains; and F_Cb is C_H2b and C_H3b immunoglobulin heavy chain constant domains.

Embodiment 2. The binding protein of embodiment 1, further comprising L₁, a linker positioned between C_H1 and V_HHa on the third polypeptide chain, and L₂, a linker positioned between V_HHa and the F_Ca on the third polypeptide chain, wherein L₁ and L₂ are each independently a linker or are absent.

Embodiment 3. The binding protein of either embodiment 1 or embodiment 2, further comprising L₃, a linker positioned between C_H1 and V_HHb on the fourth polypeptide chain, and L₄, a linker positioned between V_HHb and F_Cb on the fourth polypeptide chain, wherein L₃ and L₄ are each independently a linker or are absent.

Embodiment 4. The binding protein of any one of embodiments 1-3 further comprising H1, an immunoglobulin hinge region positioned between C_H1 and V_HHa on the third polypeptide chain, and H₂, an immunoglobulin hinge region positioned between V_HHa and the F_Ca on the third polypeptide chain, wherein H₁ and H₂ are each independently an immunoglobulin hinge region or are absent.

Embodiment 5. The binding protein of embodiment 4, wherein H₁ comprises SEQ ID NO: 53 or DK (SEQ ID NO: 54) or SEQ ID NO: 90 or is absent, and wherein H₂ comprises SEQ ID NO: 53 or SEQ ID NO: 55 or SEQ ID NO: 90 or is absent.

Embodiment 6. The binding protein of any one of embodiments 1-5, further comprising H₃, an immunoglobulin hinge region positioned between C_H1 and V_HHb on the fourth polypeptide chain, and H4, an immunoglobulin hinge region positioned between V_HHb and the F_Cb on the fourth polypeptide chain, wherein H₃ and H₄ are each independently an immunoglobulin hinge region or are absent.

Embodiment 7. The binding protein of embodiment 6, wherein H₃ comprises SEQ ID NO: 53 or DK (SEQ ID NO: 54) or SEQ ID NO: 90 or is absent, and wherein H₄ comprises SEQ ID NO: 53 or SEQ ID NO: 55 or SEQ ID NO: 90 or is absent.

Embodiment 8. The binding protein of any one of embodiments 1-7, wherein the first and second polypeptide chains have a structure represented by the formula: V_L-C_L a third polypeptide chain has a structure represented by the formula: V_H1-C_H1-H₁-L₁-V_HHa-H₂-L₂-F_Ca or V_H1-C_H1-H₁-L₁-V_HHa-L₂-H₂-F_Ca and a fourth polypeptide chain has a structure represented by the formula: V_H1-C_H1-H₃-L₃-V_HHb-H₄-L₄-F_Cb or V_H1-C_H1-H₃-L₃-V_HHb-L₄-H₄-F_Cb.

Embodiment 9. The binding protein of any one of embodiments 1-8, wherein F_Ca and/or F_Cb is from an IgG antibody.

Embodiment 10. The binding protein of embodiment 9, wherein the F_Ca and/or the F_Cb is from an IgG1 antibody.

Embodiment 11. The binding protein of any one of embodiments 1-10, wherein the binding protein activates T cells only when bound to a tumor associated antigen at one or both of the antigen binding sites.

Embodiment 12. The binding protein of any one of embodiments 2-11, wherein L₁, L₂, L₃, and/or L₄ comprise one or more repeats of the amino acid sequence of SEQ ID NO: 37, SEQ ID NO: 38 and/or SEQ ID NO: 111.

Embodiment 13. The binding protein of any one of embodiments 1-12, wherein the V_HHa comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2) and a heavy chain CDR3 (HCDR3), comprising the amino acid sequences of SEQ ID NO: 39, SEQ ID NO: 40 and SEQ ID NO: 41, respectively.

Embodiment 14. The binding protein of any one of embodiments 1-12, wherein the V_HHa comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2) and a heavy chain CDR3 (HCDR3) comprising the amino acid sequence of SEQ ID NO: 62, SEQ ID NO: 40 and SEQ ID NO: 41, respectively.

Embodiment 15. The binding protein of any one of embodiments 1-14, wherein the V_HHa comprises the amino acid sequence at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 42-48, or any one of SEQ ID NOs: 42-48.

Embodiment 16. The binding protein of any one of embodiments 1-15, wherein the T cell costimulatory molecule is CD8.

Embodiment 17. The binding protein of any one of embodiments 1-16, wherein the V_HHb comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2) and a heavy chain CDR3 (HCDR3) comprising the amino acid sequences of SEQ ID NO: 49, SEQ ID NO: 50 and SEQ ID NO: 51, respectively.

Embodiment 18. The binding protein of any one of embodiments 1-17, wherein the V_HHb comprises an amino acid sequence at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 52, or SEQ ID NO: 52.

Embodiment 19. The binding protein of any one of embodiments 1-18, wherein C_H3a and C_H3b in each of F_Ca and F_Cb comprise modifications to facilitate heterodimerization of F_Ca and F_Cb.

Embodiment 20. The binding protein of embodiment 19, wherein the modifications are substitutions to generate a knob in one of F_Ca and F_Cb and a hole in the other of F_Ca and F_Cb, wherein the knob is a substitution to tryptophan at position 366, and wherein the hole is a substitution of one or more of the following: (i) a substitution to valine at position 407; (ii) a substitution to serine at position 366; and (iii) a substitution to alanine at position 368.

Embodiment 21. The binding protein of embodiment 20, wherein the F_Ca or the F_Cb containing the knob further comprises a cysteine at position 354 and/or the F_Ca or the F_Cb containing the hole comprises a cysteine at position 349.

Embodiment 22. The binding protein of any one of embodiments 1-21 wherein the C_H2a and C_H2b immunoglobulin heavy chain constant domains each comprise the following substitutions: E233P/L234V/L235A/G236del/S267K.

Embodiment 23. The binding protein of any one of embodiments 1-22, wherein either the C_H3a immunoglobulin heavy chain constant domain or C_H3b immunoglobulin heavy chain constant domain comprises a H435R and Y436F substitution.

Embodiment 24. The binding protein of any one of embodiments 1-23, wherein F_Ca comprises the amino acid sequence of SEQ ID NO: 56 or SEQ ID NO: 57.

Embodiment 25. The binding protein of any one of embodiments 1-24, wherein F_Cb comprises the amino acid sequence of SEQ ID NO: 58.

Embodiment 26. The binding protein of any one of embodiments 1-25, wherein the third polypeptide chain comprises either SEQ ID NO: 59 or SEQ ID NO: 60 and the fourth polypeptide chain comprises SEQ ID NO: 61.

Embodiment 27. The binding protein of any one of embodiments 1-25, wherein the C_L of the first polypeptide chain and/or the C_L of the second polypeptide chain comprises a constant light chain lambda region (CLλ); and the binding protein comprises at least one lambda charge pair wherein the lambda charge pair is either between the CLλ of the first polypeptide chain and a corresponding C_H1 that is the C_H1 of the third polypeptide chain, or the lambda charge pair is between the CLλ of the second polypeptide and a corresponding C_H1 that is the C_H1 of the fourth polypeptide chain, and where the lambda charge pair is located at one or more of the following pairs of positions: (i) position 117 in the CLλ and position 141 in the corresponding C_H1; (ii) position 117 in the CLλ and position 185 in the corresponding C_H1; (iii) position 119 in the CLλ and position 128 in the corresponding C_H1; (iv) position 134 in the CLλ and position 128 in the corresponding C_H1; (v) position 134 in the CLλ and position 145 in the corresponding C_H1; (vi) position 134 in the CLλ and position 183 in the corresponding C_H1; (vii) position 136 in the CLλ and position 185 in the corresponding C_H1; (viii) position 178 in the CLλ and position 173 in the corresponding C_H1; and (ix) position 117 in the CLλ and position 187 in the corresponding C_H1; wherein the lambda charge pair comprises a positively charged amino acid residue selected from arginine, lysine and histidine located at one of the positions in the lambda charge pair and a negatively charged amino acid residue selected from aspartic acid, glutamic acid, serine and threonine located at the other position in the lambda charge pair; and wherein the numbering is according to the EU index.

Embodiment 28. The binding protein of embodiment 27, wherein the lambda charge pair is selected from the following list: (a) arginine at position 117 of the CLλ and aspartic acid at position 141 in the corresponding C_H1; (b) arginine at position 117 of the CLλ and glutamic acid at position 141 in the corresponding C_H1; (c) arginine at position 117 of the CLλ and serine at position 141 in the corresponding C_H1; (d) arginine at position 117 of the CLλ and threonine at position 141 in the corresponding C_H1; (e) lysine at position 117 of the CLλ and aspartic acid at position 141 in the corresponding C_H1; (f) lysine at position 117 of the CLλ and glutamic acid at position 141 in the corresponding C_H1; (g) lysine at position 117 of the CLλ and serine at position 141 in the corresponding C_H1; and (h) lysine at position 117 of the CLλ and threonine at position 141 in the corresponding C_H1.

Embodiment 29. The binding protein of either embodiment 27 or embodiment 28, wherein the CLλ of the first polypeptide chain and/or CLλ of the second polypeptide chain each comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NOs: 98-99.

Embodiment 30. The binding protein of any one of embodiments 1-29, wherein the C_L of the first polypeptide chain and/or the C_L of the second polypeptide chain comprises a constant light chain kappa region (CLκ); and a kappa charge pair wherein the kappa charge pair is either between the CLκ of the first polypeptide chain and a corresponding C_H1 that is the C_H1 of the third polypeptide chain, or the kappa charge pair is between the CLκ of the second polypeptide and a corresponding C_H1 that is the C_H1 of the fourth polypeptide chain; wherein the kappa charge pair is located at position 117 in the CLκ and position 141 in the corresponding C_H1 and comprises a positively charged amino acid residue selected from arginine, lysine, and histidine located at one of the positions in the kappa charge pair, and a negatively charged amino acid residue selected from aspartic acid, glutamic acid, serine and threonine located at the other position in the kappa charge pair.

Embodiment 31. The binding protein of embodiment 30, wherein the negatively charged amino acid residue in the kappa charge pair is located at position 133 of the CLκ, and the positively charged amino acid residue is located at position 183 of the corresponding C_H1, optionally wherein the negatively charged amino acid residue at position 133 of the CLκ is a glutamic acid, and optionally wherein the positively charged amino acid residue at position 183 of the corresponding C_H1 is a lysine.

Embodiment 32. The binding protein of either embodiment 30 or embodiment 31, wherein the CLκ of the first polypeptide chain or the second polypeptide chain comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 100.

Embodiment 33. The binding protein of any one of embodiments 30-32 wherein the corresponding CH1 in each of the one or more lambda charge pairs comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to SEQ ID NO: 95-97, and/or the corresponding C_H1 in the kappa charge pair comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to SEQ ID NO: 100.

Embodiment 34. The binding protein of any one of embodiments 27-33, wherein the C_L, comprises an amino acid sequence according to SEQ ID NO: 105, and the corresponding C_H1 comprises an amino acid sequence according to SEQ ID NO: 104.

Embodiment 35. The binding protein of any one of embodiments 1-34, wherein either: (i) a disulfide link between the C_L of the first polypeptide chain and the C_H1 of the third polypeptide chain is formed between a pair of cysteines engineered into the C_L of the first polypeptide chain and the C_H1 of the third polypeptide chain, and a disulfide link between the C_L of the second polypeptide chain and the C_H1 of the fourth polypeptide chain is formed between a pair of native cysteines; or (ii) a disulfide link between the C_L of the second polypeptide chain and the C_H1 of the fourth polypeptide chain is formed between a pair of cysteines engineered into the C_L of the second polypeptide chain and the C_H1 of the fourth polypeptide chain, and a disulfide link between the C_L of the first polypeptide chain and the C_H1 of the third polypeptide chain is formed between a pair of native cysteines.

Embodiment 36. The binding protein of embodiment 35, wherein the pair of cysteines are engineered into either: (i) the C_L of the first polypeptide chain and the C_H1 of the third polypeptide chain, wherein the C_L of the first polypeptide chain is a CLλ and wherein the pair of engineered cysteines are located at position 122 of the CLλ of the first polypeptide chain and position 126 of the C_H1 of the third polypeptide chain, and wherein the C_L, of the first polypeptide chain comprises a non-cysteine residue at position 212 and the C_H1 of the third polypeptide chain comprises a non-cysteine residue at position 220, optionally wherein the non-cysteine residues are valines; or (ii) the C_L of the second polypeptide chain and the C_H1 of the fourth polypeptide chain, wherein the C_L of the first polypeptide chain is a CLλ and wherein the pair of engineered cysteines are located at position 122 of the CLλ of the second polypeptide chain and position 126 of the C_H1 of the fourth polypeptide chain, and wherein the CLλ of the second polypeptide chain comprises a non-cysteine residue at position 212 and the C_H1 of the fourth polypeptide chain comprises a non-cysteine residue at position 220, optionally wherein the non-cysteine residues are valines.

Embodiment 37. The binding protein of embodiment 35, wherein the pair of cysteines are engineered into either: (i) the C_L of the first polypeptide chain and the C_H1 of the third polypeptide chain, wherein the C_L of the first polypeptide chain is a CLκ and wherein the pair of engineered cysteines are located at position 121 of the CLκ of the first polypeptide chain and position 126 of the C_H1 of the third polypeptide chain, and wherein the CLκ of the first polypeptide chain comprises a non-cysteine residue at position 214 and the C_H1 of the third polypeptide chain comprises a non-cysteine residue at position 220, optionally wherein the non-cysteine residues are valines; or (ii) the C_L of the second polypeptide chain and the C_H1 of the fourth polypeptide chain, wherein the C_L of the first polypeptide chain is a CLκ and wherein the pair of engineered cysteines are located at position 121 of the CLκ of the second polypeptide chain and position 126 of the C_H1 of the fourth polypeptide chain, and wherein the CLκ of the second polypeptide chain comprises a non-cysteine residue at position 214 and the C_H1 of the fourth polypeptide chain comprises a non-cysteine residue at position 220, optionally wherein the non-cysteine residues are valines.

Embodiment 38. The binding protein of any one of embodiments 35-37, wherein the C_L, comprises an amino acid sequence according to SEQ ID NO: 107, and the corresponding C_H1 comprises an amino acid sequence according to SEQ ID NO: 106.

Embodiment 39. The binding protein of any one of embodiments 1-38, wherein the tumor-associated antigen (TAA) is CD20, Glypican-3 (GPC3), or leucine rich repeat containing 15 (LRRC15).

Embodiment 40. The binding protein of any one of embodiments 1-39, wherein the TAA binding domain binds to CD20 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a light chain CDR3 (LCDR3), comprising the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, respectively.

Embodiment 41. The binding protein of embodiment 40, wherein the TAA binding domain comprises a V_H1 domain and a V_L domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 7 and SEQ ID NO: 8, respectively; or comprises a V_H chain according to SEQ ID NO: 7 and a light chain according to SEQ ID NO: 8.

Embodiment 42. The binding protein of any one of embodiments 1-35, wherein the TAA binding domain binds to GPC3 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, and SEQ ID NO: 18, respectively.

Embodiment 43. The binding protein of embodiment 36, wherein the TAA binding domain comprises a V_H1 domain and a V_L domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 19 and SEQ ID NO: 20, respectively; or comprises a V_H chain according to SEQ ID NO: 19 and a light chain according to SEQ ID NO: 20.

Embodiment 44. The binding protein of any one of embodiments 1-39, wherein the TAA binding domain binds to LRRC15 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO: 30 respectively.

Embodiment 45. The binding protein of embodiment 44, wherein the TAA binding domain comprises a V_H1 domain and a V_L domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 31 and SEQ ID NO: 32, respectively; or comprises a V_H chain according to SEQ ID NO: 31 and a light chain according to SEQ ID NO: 32.

Embodiment 46. The binding protein of any one of embodiments 1-45 comprising: (a) the amino acid of sequences of SEQ ID NO: 9, SEQ ID NO: 11, and SEQ ID NO: 12; (b) the amino acid of sequences of SEQ ID NO: 21, SEQ ID NO: 23, and SEQ ID NO: 24; or (c) the amino acid of sequences of SEQ ID NO: 33, SEQ ID NO: 35, and SEQ ID NO: 36.

Embodiment 47. The binding protein of embodiment 46, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 9, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 11, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 12.

Embodiment 48. The binding protein of embodiment 46, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 21, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 23, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 24.

Embodiment 49. The binding protein of embodiment 46, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 33, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 35, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 36.

Embodiment 50. A pharmaceutical composition comprising the binding protein of any one of embodiments 1-49 and a pharmaceutically acceptable carrier.

Embodiment 51. An isolated nucleic acid molecule encoding the binding protein of any one of embodiments 1-49.

Embodiment 52. A vector comprising the isolated nucleic acid molecule of embodiment 51.

Embodiment 53. A method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of embodiments 1-49 or the pharmaceutical composition of embodiment 50 to the subject.

Embodiment 54. A method of treating an inflammatory disease and/or autoimmune disorder in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of embodiments 1-49 or the pharmaceutical composition of embodiment 50 to the subject.

Embodiment 55. The method of either embodiment 53, or embodiment 54, wherein the binding protein preferentially activates a subset of T cells in the subject.

Embodiment 56. The method of embodiment 55, wherein the subset of T cells are CD8+ T cells.

Embodiment 57. The method of embodiment 56, wherein the CD8+ T cells are preferentially activated as compared to CD4+ T cells.

Embodiment 58. The method of any one of embodiments 53-57, wherein the activation of T cells is determined by measuring the percentage of surface CD25+ T cells.

Embodiment 59. The method of embodiment 58, wherein the percentage of surface CD25+ T cells that are CD8+ T cells is higher than the percentage of surface CD25+ T cells that are CD4+ T cells.

Embodiment 60. A method of treating an inflammatory disease and/or autoimmune disorder in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of embodiments 1-49 or the pharmaceutical composition of embodiment 50 to the subject.

Embodiment 61. The method of embodiment 53, wherein the tumor-associated antigen of the binding protein is CD20 and the cancer is a B-cell malignancy.

Embodiment 62. The method of embodiment 61, wherein the B-cell malignancy is non-Hodgkin's lymphoma (NHL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Burkett lymphoma, or primary mediastinal large B cell lymphoma (PMBCL).

Embodiment 63. The method of embodiment 53, wherein the tumor-associated antigen of the binding protein is Glypican-3 (GPC3) and the cancer is liver cancer, hepatocellular carcinoma (HCC), non-small cell lung cancer (NSCLC), squamous non-small cell lung cancer (sqNSCLC), ovarian cancer, clear cell ovarian cancer, carcinoma, Merkel cell carcinoma, gastric cancer, hepatoblastoma, or nephroblastoma.

Embodiment 64. The method of embodiment 53, wherein the tumor-associated antigen of the binding protein is leucine rich repeat containing 15 (LRRC15) and the cancer is melanoma, sarcoma, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial cell carcinoma, osteosarcoma, glioblastoma, lung cancer, non-small cell lung cancer, or thyroid cancer.

Embodiment 65. The method of embodiment 54, wherein the inflammatory disease and/or autoimmune disorder is systemic lupus erythematosus (SLE), myositis, rheumatoid arthritis (RA), Sjogren's syndrome, or anti-neutrophil cytoplasmic autoantibody (ANCA) vasculitis.

Embodiment 66. A method of treating systemic lupus erythematosus (SLE) in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of embodiments 1-49 or the pharmaceutical composition of embodiment 50 to the subject.

Embodiment 67. A method of treating myositis in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of embodiments 1-49 or the pharmaceutical composition of embodiment 50 to the subject.

Embodiment 68. A method of treating rheumatoid arthritis (RA) in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of embodiments 1-49 or the pharmaceutical composition of embodiment 50 to the subject.

Embodiment 69. A method of treating Sjogren's syndrome in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of embodiments 1-49 or the pharmaceutical composition of embodiment 50 to the subject.

Embodiment 70. The method of any one of embodiments 53-69, wherein the binding protein is administered by subcutaneous administration.

Embodiment 71. A binding protein of any one of embodiments 1-49, or the pharmaceutical composition of embodiment 50, for use as a medicament.

Embodiment 72. A binding protein of any one of embodiments 1-49, or the pharmaceutical composition of embodiment 50, for use in the treatment of cancer.

Embodiment 73. A binding protein of any one of embodiments 1-49, or the pharmaceutical composition of embodiment 50, for use in the treatment of an inflammatory disease and/or autoimmune disorder.

Embodiment 74. The binding protein or pharmaceutical composition for use of embodiment 72, wherein the tumor-associated antigen is CD20 and the cancer is a B-cell malignancy.

Embodiment 75. The binding protein or pharmaceutical composition for use of embodiment 74, wherein the B-cell malignancy is non-Hodgkin's lymphoma (NHL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Burkett lymphoma, or primary mediastinal large B cell lymphoma (PMBCL).

Embodiment 76. The binding protein or pharmaceutical composition for use of embodiment 72, wherein the tumor-associated antigen is Glypican-3 (GPC3) and the cancer is liver cancer or hepatocellular carcinoma (HCC), non-small cell lung cancer (NSCLC), squamous non-small cell lung cancer (sqNSCLC), ovarian cancer, clear cell ovarian cancer, carcinoma, Merkel cell carcinoma, gastric cancer, hepatoblastoma, or nephroblastoma.

Embodiment 77. The binding protein or pharmaceutical composition for use of embodiment 72, wherein the tumor-associated antigen is leucine rich repeat containing 15 (LRRC15) and the cancer is melanoma, sarcoma, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial cell carcinoma, osteosarcoma, glioblastoma, lung cancer, non-small cell lung cancer, or thyroid cancer.

Embodiment 78. The binding protein or pharmaceutical composition for use of embodiment 73, wherein the inflammatory disease and/or autoimmune disorder is systemic lupus erythematosus (SLE), myositis, rheumatoid arthritis (RA), Sjogren's syndrome, or ANCA vasculitis.

Embodiment 79. A binding protein of any one of embodiments 1-49, or the pharmaceutical composition of embodiment 50, for use in the treatment of systemic lupus erythematosus (SLE).

Embodiment 80. A binding protein of any one of embodiments 1-49, or the pharmaceutical composition of embodiment 50, for use in the treatment of myositis.

Embodiment 81. A binding protein of any one of embodiments 1-49, or the pharmaceutical composition of embodiment 50, for use in the treatment of rheumatoid arthritis (RA).

Embodiment 82. A binding protein of any one of embodiments 1-49, or the pharmaceutical composition of embodiment 50, for use in the treatment of Sjogren's syndrome.

Embodiment 83. Use of a binding protein of any one of embodiments 1-49, or the pharmaceutical composition of embodiment 50, for the manufacture of a medicament for treating cancer.

Embodiment 84. Use of a binding protein of any one of embodiments 1-49, or the pharmaceutical composition of embodiment 50, for the manufacture of a medicament for treating an inflammatory and/or autoimmune disorder.

Embodiment 85. The use of embodiment 83, wherein the cancer is a B-cell malignancy, liver cancer, or HCC.

Embodiment 86. The use of embodiment 85, wherein the B-cell malignancy is non-Hodgkin's lymphoma (NHL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Burkett lymphoma, or primary mediastinal large B cell lymphoma (PMBCL).

Embodiment 87. The use of embodiment 83, wherein the cancer is melanoma, sarcoma, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial cell carcinoma, osteosarcoma, glioblastoma, lung cancer, non-small cell lung cancer, or thyroid cancer.

Embodiment 88. The use of embodiment 84, wherein the inflammatory and/or autoimmune disorder is systemic lupus erythematosus (SLE), myositis, rheumatoid arthritis (RA), Sjogren's syndrome, or ANCA vasculitis.

Embodiment 89. Use of a binding protein of any one of embodiments 1-49, or the pharmaceutical composition of embodiment 50, for the manufacture of a medicament for the treatment of systemic lupus erythematosus (SLE).

Embodiment 90. Use of a binding protein of any one of embodiments 1-49, or the pharmaceutical composition of embodiment 50, for the manufacture of a medicament for the treatment of myositis.

Embodiment 91. Use of a binding protein of any one of embodiments 1-49, or the pharmaceutical composition of embodiment 50, for the manufacture of a medicament for the treatment of rheumatoid arthritis (RA).

Embodiment 92. Use of a binding protein of any one of embodiments 1-49, or the pharmaceutical composition of embodiment 50, for the manufacture of a medicament for the treatment of Sjogren's syndrome.

EXAMPLES

The Examples that follow are illustrative of specific aspects of the disclosure, and various uses thereof. They are set forth for explanatory purposes only and should not be construed as limiting the scope of the disclosure in any way.

Example 1: Binding Protein Structures and Sequences

A novel class of binding proteins engineered to improve safety and increase efficacy is described herein. A multitude of binding protein formats were tested to determine the placement of the TCR binding domain, the placement of the T-cell co-stimulatory domain, the linker size and cleavability and the Fc portion to be used.

TABLE 1
Amino acid and Nucleic acid sequences for LM1486 (CD20), LM1653 (GPC3),
LRC150016 (LRRC15), TPP-46956-TPP-46960 (LRRC15), TPP-42197 (BCMA) and
LM1953 (STEAP-2)
SEQ ID
NO:	Description	Sequence
1	LM1486/	GYTFTSYNMH
	LM1486-2 CD20
	VH CDR1
2	LM1486/	AIYPGSGDTSYNQKFKG
	LM1486-2
	CD20 VH
	CDR2
3	LM1486/	VVYYSNSYWYFDV
	LM1486-2
	CD20 VH
	CDR3
4	LM1486/	RASSSVSYMH
	LM1486-2
	CD20 VL
	CDR1
5	LM1486/	APSNLAS
	LM1486-2
	CD20 VL
	CDR2
6	LM1486/	QQWSFNPPT
	LM1486-2
	CD20 VL
	CDR3
7	LM1486/	EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYNMHWVRQA
	LM1486-2	PGKGLEWVGAIYPGSGDTSYNQKFKGRFTISVDKSKNTLYL
	CD20 VH	QMNSLRAEDTAVYYCARVVYYSNSYWYFDVWGQGTLVTV
		SS
8	LM1486/	DIQMTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGK
	LM1486-2	APKPLIYAPSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFATY
	CD20 VL	YCQQWSFNPPTFGQGTKVEIK
9	LM1486/	DIQMTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGK
	LM1486-2	APKPLIYAPSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFATY
	CD20 Light	YCQQWSFNPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGT
	Chain	ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSK
		DSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR
		GEC
10	LM1486/	EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYNMHWVRQA
	LM1486-2	PGKGLEWVGAIYPGSGDTSYNQKFKGRFTISVDKSKNTLYL
	CD20 VH-	QMNSLRAEDTAVYYCARVVYYSNSYWYFDVWGQGTLVTV
	CH1	SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS
		WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
		NVNHKPSNTKVDKRVEPKSC
11	LM1486-2-	EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYNMHWVRQA
	hole (TCR)	PGKGLEWVGAIYPGSGDTSYNQKFKGRFTISVDKSKNTLYL
	CD20 HC	QMNSLRAEDTAVYYCARVVYYSNSYWYFDVWGQGTLVTV
	TCR (FC	SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS
	hole)	WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
		NVNHKPSNTKVDKRVEPKSCDKTGGSEVQLVESGGGLVQPG
		GSLRLSCVASGDVHKINFLGWYRQAPGKEREKVAHISIGDQT
		DYADSAKGRFTISRDESKNMVYLQMNSLKPEDTAVYFCRAF
		SRIYPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVF
		LFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGV
		EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
		VSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSL
		SCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV
		SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
12	LM1486/	EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYNMHWVRQA
	LM1486-2-	PGKGLEWVGAIYPGSGDTSYNQKFKGRFTISVDKSKNTLYL
	knob (CD8)	QMNSLRAEDTAVYYCARVVYYSNSYWYFDVWGQGTLVTV
	CD20 HC	SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS
	CD8 (FC	WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
	knob)	NVNHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPG
		GSLRLSCAASGFTFDDYAIGWFRQAPGKEREGVSCIRVSDGS
		TYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAA
		GSLYTCVQSIVWPARPYYDMDYWGQGTLVTVSSGGGGSTH
		TCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKH
		EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV
		LHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY
		TLPPCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENN
		YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA
		LHNHYTQKSLSLSPGK
13	LM1653	DYEMH
	GPC3 VH
	CDR1
14	LM1653	ALDPKTGDTAYSQKFKG
	GPC3 VH
	CDR2
15	LM1653	FYSYTY
	GPC3 VH
	CDR3
16	LM1653	RSSQSLVHSNRNTYLH
	GPC3 VL
	CDR1
17	LM1653	KVSNRFS
	GPC3 VL
	CDR2
18	LM1653	SQNTHVPPT
	GPC3 VL
	CDR3
19	LM1653	QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYEMHWVRQ
	GPC3 VH	APGQGLEWMGALDPKTGDTAYSQKFKGRVTLTADKSTSTA
		YMELSSLTSEDTAVYYCTRFYSYTYWGQGTLVTVSS
20	LM1653	DVVMTQSPLSLPVTPGEPASISCRSSQSLVHSNRNTYLHWYL
	GPC3 VL	QKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEA
		EDVGVYYCSQNTHVPPTFGQGTKLEIK
21	LM1653	DVVMTQSPLSLPVTPGEPASISCRSSQSLVHSNRNTYLHWYL
	GPC3 Light	QKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEA
	Chain	EDVGVYYCSQNTHVPPTFGQGTKLEIKRTVAAPSVFIFPPSDE
		QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV
		TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV
		TKSFNRGEC
22	LM1653	QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYEMHWVRQ
	GPC3 VH-	APGQGLEWMGALDPKTGDTAYSQKFKGRVTLTADKSTSTA
	CH1	YMELSSLTSEDTAVYYCTRFYSYTYWGQGTLVTVSSASTKG
		PSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT
		SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS
		NTKVDKRVEPKSC
23	LM1653	QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYEMHWVRQ
	GPC3-hole	APGQGLEWMGALDPKTGDTAYSQKFKGRVTLTADKSTSTA
	(TCR)	YMELSSLTSEDTAVYYCTRFYSYTYWGQGTLVTVSSASTKG
	HC TCR (FC	PSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT
	hole)	SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS
		NTKVDKRVEPKSCDKTGGSEVQLVESGGGLVQPGGSLRLSC
		VASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTDYADSA
		KGRFTISRDESKNMVYLQMNSLKPEDTAVYFCRAFSRIYPYD
		YWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPPKPK
		DTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVHNAK
		TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL
		PAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCAVKG
		FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVD
		KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
24	LM1653	QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYEMHWVRQ
	GPC3-knob	APGQGLEWMGALDPKTGDTAYSQKFKGRVTLTADKSTSTA
	(CD8)	YMELSSLTSEDTAVYYCTRFYSYTYWGQGTLVTVSSASTKG
	HC CD8 (FC	PSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT
	knob)	SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS
		NTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGSLRLSC
		AASGFTFDDYAIGWFRQAPGKEREGVSCIRVSDGSTYYADS
		VKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAAGSLYTC
		VQSIVWPARPYYDMDYWGQGTLVTVSSGGGGSTHTCPPCP
		APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV
		KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
		WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPC
		REEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTP
		PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
		TQKSLSLSPGK
25	LRC150016	SYWIE
	VH CDR1
26	LRC150016	EILPGSDTTNYNEKFKD
	VH CDR2
27	LRC150016	DRGNYRAWFGY
	VH CDR3
28	LRC150016	RASQDISNYLN
	VL CDR1
29	LRC150016	YTSRLHS
	VL CDR2
30	LRC150016	QQGEALPWT
	VL CDR3
31	LRC150016	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	VH	PGQGLEWIGEILPGSDTTNYNEKFKDRATFTSDTSINTAYME
		LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSS
32	LRC150016	DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQKPGG
	VL	AVKFLIYYTSRLHSGVPSRFSGSGSGTDYTLTISSLQPEDFAT
		YFCQQGEALPWTFGGGTKVEIK
33	LRC150016	DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQKPGG
	Light Chain	AVKFLIYYTSRLHSGVPSRFSGSGSGTDYTLTISSLQPEDFAT
		YFCQQGEALPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKS
		GTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD
		SKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSF
		NRGEC
34	LRC150016	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	VH-CHI	PGQGLEWIGEILPGSDTTNYNEKFKDRATFTSDTSINTAYME
		LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAS
		TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
		ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
		KPSNTKVDKRVEPKSC
35	LRC150016-	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	hole (TCR)	PGQGLEWIGEILPGSDTTNYNEKFKDRATFTSDTSINTAYME
	HC (FC hole)	LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAS
		TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
36	LRC150016-	ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
	knob (CD8)	KPSNTKVDKRVEPKSCDKTGGSEVQLVESGGGLVQPGGSLR
	HC CD8 (FC	LSCVASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTDYA
	knob)	DSAKGRFTISRDESKNMVYLQMNSLKPEDTAVYFCRAFSRIY
		PYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPP
		KPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVH
		NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
		KALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCA
		VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKL
		TVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
		EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
		PGQGLEWIGEILPGSDTTNYNEKFKDRATFTSDTSINTAYME
		LSRLRSDDTAVYYCARDRGNYRAWEGYWGQGTLVTVSSAS
		TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
		ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
		KPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGSLR
		LSCAASGFTFDDYAIGWFRQAPGKEREGVSCIRVSDGSTYYA
		DSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAAGSLY
		TCVQSIVWPARPYYDMDYWGQGTLVTVSSGGGGSTHTCPP
		CPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDP
		EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ
		DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPP
		CREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT
		PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
		YTQKSLSLSPGK
37	linker	TGGS
38	linker	GGGGS
39	TCR VHHa	INFLG
	CDR1
40	TCR VHHa	HISIGDQTDYADSAKG
	CDR2
41	TCR VHHa	FSRIYPYDY
	CDR3
42	TCR VHHa	EVQLVESGGGLVQPGGSLRLSCVASGDVHKINFLGWYRQAP
		GKEREKVAHISIGDQTDYADSAKGRFTISRDESKNMVYLQM
		NSLKPEDTAVYFCRAFSRIYPYDYWGQGTLVTVSS
43	TCR hu2.1	EVQLLESGGGLVQPGGSLRLSCAASGDVHKINFLSWYRQAP
	VHHa	GKEREKVAHISIGDQTDYADSAKGRFTISRDESKNTLYLQMN
		SLRAEDTAVYYCRAFSRIYPYDYWGQGTLVTVSS
44	TCR hu2.2	EVQLLESGGGLVQPGGSLRLSCAASGDVHKINFLGWYRQAP
	VHHa	GKEREKVAHISIGDQTDYADSAKGRFTISRDESKNTLYLQMN
		SLRAEDTAVYYCRAFSRIYPYDYWGQGTLVTVSS
45	TCR hu2.3	EVQLLESGGGLVQPGGSLRLSCAASGDVHKINFLSWYRQAP
	VHHa	GKGLEKVAHISIGDQTDYADSAKGRFTISRDESKNTLYLQMN
		SLRAEDTAVYYCRAFSRIYPYDYWGQGTLVTVSS
46	TCR hu2.4	EVQLLESGGGLVQPGGSLRLSCAASGDVHKINFLGWYRQAP
	VHHa	GKGLEKVAHISIGDQTDYADSAKGRFTISRDESKNTLYLQMN
		SLRAEDTAVYYCRAFSRIYPYDYWGQGTLVTVSS
47	TCR hu2.5	EVQLLESGGGLVQPGGSLRLSCAASGDVHKINFLGWYRQAP
	VHHa	GKEREKVAHISIGDQTDYADSAKGRFTISRDESKNTVYLQMN
		SLRAEDTAVYFCRAFSRIYPYDYWGQGTLVTVSS
48	TCR hu2.6	EVQLLESGGGLVQPGGSLRLSCAASGDVHKINFLGWYRQAP
	VHHa	GKGLEKVAHISIGDQTDYADSAKGRFTISRDESKNTVYLQM
		NSLRAEDTAVYFCRAFSRIYPYDYWGQGTLVTVSS
49	CD8 VHHb	DYAIG
	CDR1
50	CD8 VHHb	IRVSDGSTYYADSVKG
	CDR2
51	CD8 VHHb	GSLYTCVQSIVWPARPYYDMDY
	CDR3
52	CD8 VHHb	EVQLLESGGGLVQPGGSLRLSCAASGFTFDDYAIGWFRQAP
	LM1486 (CD20	GKEREGVSCIRVSDGSTYYADSVKGRFTISRDNSKNTLYLQM
	CD8)	NSLRAEDTAVYYCAAGSLYTCVQSIVWPARPYYDMDYWGQ
	BCMA CD8	GTLVTVSS
	LM1953
	(STEAP2
	CD8)
	LRC150016
	(LRRC15)
53	IgG1 hinge	DKTHTCPPCPAP
	sequence
54	IgG1 Split	DK
	Hinge 1
55	IgG1 Split	THTCPPCPAP
	Hinge 2
56	Fca (hole)	PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKF
		NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
		NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREE
		MTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVL
		DSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ
		KSLSLSPGK
57	Fca (hole)	PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKF
	(RF)	NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
		NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREE
		MTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVL
		DSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNRFTQK
		SLSLSPGK
58	Fcb (knob)	PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKF
		NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
		NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRE
		EMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPV
		LDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT
		QKSLSLSPGK
59	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
		NHKPSNTKVDKRVEPKSCDKTGGSEVQLVESGGGLVQPGGS
		LRLSCVASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTD
		YADSAKGRFTISRDESKNMVYLQMNSLKPEDTAVYFCRAFS
		RIYPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFL
		FPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVE
		VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
		SNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLS
		CAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVS
		KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
60	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole/RF)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
		NHKPSNTKVDKRVEPKSCDKTGGSEVQLVESGGGLVQPGGS
		LRLSCVASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTD
		YADSAKGRFTISRDESKNMVYLQMNSLKPEDTAVYFCRAFS
		RIYPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFL
		FPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVE
		VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
		SNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLS
		CAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVS
		KLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
61	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(CD8/knob)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
		NHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGS
		LRLSCAASGFTEDDYAIGWFRQAPGKEREGVSCIRVSDGSTY
		YADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAAGS
		LYTCVQSIVWPARPYYDMDYWGQGTLVTVSSGGGGSTHTC
		PPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHED
		PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH
		QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL
		PPCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYK
		TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH
		NHYTQKSLSLSPGK
62	TCR VHHa hu	INFLS
	2.1, 2.3
	CDR1
63	LRC150016-	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	hole (TCR)	PGQGLEWIGEILPGSDTTNYNEKFKDRATFTSDTSINTAYME
	HC TCR (FC	LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAS
	hole) hu2.1	TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
	VHHa	ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
		KPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGSLR
		LSCAASGDVHKINFLSWYRQAPGKEREKVAHISIGDQTDYA
		DSAKGRFTISRDESKNTLYLQMNSLRAEDTAVYYCRAFSRIY
		PYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPP
		KPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVH
		NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
		KALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCA
		VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKL
		TVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
64	LRC150016-	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	hole (TCR)	PGQGLEWIGEILPGSDTTNYNEKFKDRATFTSDTSINTAYME
	HC TCR (FC	LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAS
	hole) hu2.2	TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
	VHHa	ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
		KPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGSLR
		LSCAASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTDYA
		DSAKGRFTISRDESKNTLYLQMNSLRAEDTAVYYCRAFSRIY
		PYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPP
		KPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVH
		NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
		KALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCA
		VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKL
		TVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
65	LRC150016-	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	hole (TCR)	PGQGLEWIGEILPGSDTTNYNEKFKDRATFTSDTSINTAYME
	HC TCR (FC	LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAS
	hole) hu2.3	TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
	VHHa	ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
		KPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGSLR
		LSCAASGDVHKINFLSWYRQAPGKGLEKVAHISIGDQTDYA
		DSAKGRFTISRDESKNTLYLQMNSLRAEDTAVYYCRAFSRIY
		PYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPP
		KPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVH
		NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
		KALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCA
		VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKL
		TVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
66	LRC150016-	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	hole (TCR)	PGQGLEWIGEILPGSDTTNYNEKFKDRATFTSDTSINTAYME
	HC TCR (FC	LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAS
	hole) hu2.4	TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
	VHHa	ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
		KPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGSLR
		LSCAASGDVHKINFLGWYRQAPGKGLEKVAHISIGDQTDYA
		DSAKGRFTISRDESKNTLYLQMNSLRAEDTAVYYCRAFSRIY
		PYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPP
		KPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVH
		NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
		KALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCA
		VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKL
		TVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
67	LRC150016-	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	hole (TCR)	PGQGLEWIGEILPGSDTTNYNEKFKDRATFTSDTSINTAYME
	HC TCR (FC	LSRLRSDDTAVYYCARDRGNYRAWEGYWGQGTLVTVSSAS
	hole) hu2.5	TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
	VHHa	ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
		KPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGSLR
		LSCAASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTDYA
		DSAKGRFTISRDESKNTVYLQMNSLRAEDTAVYFCRAFSRIY
		PYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPP
		KPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVH
		NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
		KALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCA
		VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKL
		TVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
68	LRC150016-	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	hole (TCR)	PGQGLEWIGEILPGSDTTNYNEKFKDRATFTSDTSINTAYME
	HC TCR (FC	LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAS
	hole) hu2.6	TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
	VHHa	ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
		KPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGSLR
		LSCAASGDVHKINFLGWYRQAPGKGLEKVAHISIGDQTDYA
		DSAKGRFTISRDESKNTVYLQMNSLRAEDTAVYFCRAFSRIY
		PYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPP
		KPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVH
		NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
		KALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCA
		VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKL
		TVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
69	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
	TCR hu2.1	NHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGS
	VHHa	LRLSCAASGDVHKINFLSWYRQAPGKEREKVAHISIGDQTDY
		ADSAKGRFTISRDESKNTLYLQMNSLRAEDTAVYYCRAFSRI
		YPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFP
		PKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEV
		HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS
		NKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSC
		AVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSK
		LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
70	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
	TCR hu2.2	NHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGS
	VHHa	LRLSCAASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTD
		YADSAKGRFTISRDESKNTLYLQMNSLRAEDTAVYYCRAFS
		RIYPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFL
		FPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVE
		VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
		SNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLS
		CAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVS
		KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
71	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
	TCR hu2.3	NHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGS
	VHHa	LRLSCAASGDVHKINFLSWYRQAPGKGLEKVAHISIGDQTDY
		ADSAKGRFTISRDESKNTLYLQMNSLRAEDTAVYYCRAFSRI
		YPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFP
		PKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEV
		HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS
		NKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSC
		AVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSK
		LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
72	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
	TCR hu2.4	NHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGS
	VHHa	LRLSCAASGDVHKINFLGWYRQAPGKGLEKVAHISIGDQTD
		YADSAKGRFTISRDESKNTLYLQMNSLRAEDTAVYYCRAFS
		RIYPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFL
		FPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVE
		VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
		SNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLS
		CAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVS
		KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
73	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
	TCR hu2.5	NHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGS
	VHHa	LRLSCAASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTD
		YADSAKGRFTISRDESKNTVYLQMNSLRAEDTAVYFCRAFS
		RIYPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFL
		FPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVE
		VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
		SNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLS
		CAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVS
		KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
74	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
	TCR hu2.6	NHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGS
	VHHa	LRLSCAASGDVHKINFLGWYRQAPGKGLEKVAHISIGDQTD
		YADSAKGRFTISRDESKNTVYLQMNSLRAEDTAVYFCRAFS
		RIYPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFL
		FPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVE
		VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
		SNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLS
		CAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVS
		KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
75	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole/RF)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
	TCR hu2.1	NHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGS
	VHHa	LRLSCAASGDVHKINFLSWYRQAPGKEREKVAHISIGDQTDY
		ADSAKGRFTISRDESKNTLYLQMNSLRAEDTAVYYCRAFSRI
		YPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFP
		PKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEV
		HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS
		NKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSC
		AVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSK
		LTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
76	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole/RF)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
	TCR hu2.2	NHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGS
	VHHa	LRLSCAASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTD
		YADSAKGRFTISRDESKNTLYLQMNSLRAEDTAVYYCRAFS
		RIYPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFL
		FPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVE
		VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
		SNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLS
		CAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVS
		KLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
77	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole/RF)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
	TCR hu2.3	NHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGS
	VHHa	LRLSCAASGDVHKINFLSWYRQAPGKGLEKVAHISIGDQTDY
		ADSAKGRFTISRDESKNTLYLQMNSLRAEDTAVYYCRAFSRI
		YPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFP
		PKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEV
		HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS
		NKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSC
		AVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSK
		LTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
78	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole/RF)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
	TCR hu2.4	NHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGS
	VHHa	LRLSCAASGDVHKINFLGWYRQAPGKGLEKVAHISIGDQTD
		YADSAKGRFTISRDESKNTLYLQMNSLRAEDTAVYYCRAFS
		RIYPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVEL
		FPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVE
		VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
		SNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLS
		CAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVS
		KLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
79	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole/RF)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
	TCR hu2.5	NHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGS
	VHHa	LRLSCAASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTD
		YADSAKGRFTISRDESKNTVYLQMNSLRAEDTAVYFCRAFS
		RIYPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFL
		FPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVE
		VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
		SNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLS
		CAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVS
		KLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
80	Backbone	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN
	(TCR/hole/RF)	SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
	TCR hu2.6	NHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGS
	VHHa	LRLSCAASGDVHKINFLGWYRQAPGKGLEKVAHISIGDQTD
		YADSAKGRFTISRDESKNTVYLQMNSLRAEDTAVYFCRAFS
		RIYPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFL
		FPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVE
		VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
		SNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLS
		CAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVS
		KLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
81	CD8 VHHb	EVQLLESGGGLVQPGGSLRLSCAASGFTFPDYAIGWFRQAPG
	D30P/S75A/	KEREGVSCIRVSDGSTYYADSVKGRFTISRDNAKNTLYLQM
	Q106N	NSLRAEDTAVYYCAAGSLYTCVNSIVWPARPYYDMDYWGQ
		GTLVTVSS
82	CD8 VHHb	EVQLLESGGGLVQPGGSLRLSCAASGFTFPDYAIGWFRQAPG
	D30P/S75A/	KEREGVSCIRVSDGSTYYADSVKGRFTISRDNAKNTLYLQM
	L101A	NSLRAEDTAVYYCAAGSAYTCVQSIVWPARPYYDMDYWG
		QGTLVTVSS
83	CD8 VHHb	EVQLLESGGGLVQPGGSLRLSCAASGFTFRDYAIGWFRQAPG
	S75A/D30R	KEREGVSCIRVSDGSTYYADSVKGRFTISRDNAKNTLYLQM
		NSLRAEDTAVYYCAAGSLYTCVQSIVWPARPYYDMDYWGQ
		GTLVTVSS
84	CD8 VHHb	EVQLLESGGGLVQPGGSLRLSCAASGFTFPDYAIGWFRQAPG
	D30P/S75A/	KEREGVSCIRVSDGSTYYADSVKGRFTISRDNAKNTLYLQM
	S100G	NSLRAEDTAVYYCAAGGLYTCVQSIVWPARPYYDMDYWG
		QGTLVTVSS
85	CD8 VHHb	EVQLLESGGGLVQPGGSLRLSCAASGFTFPDYAIGWFRQAPG
	D30P/S75A/	KEREGVSCIRVSDGSTYYADSVKGRFTISRDNAKNTLYLQM
	Y102I	NSLRAEDTAVYYCAAGSLITCVQSIVWPARPYYDMDYWGQ
		GTLVTVSS
86	CD8 VHHb	EVQLLESGGGLVQPGGSLRLSCAASGFTFPDYAIGWFRQAPG
	D30P/S75A/	KEREGVSCIRVSDGSTYYADSVKGRFTISRDNAKNTLYLQM
	S100P	NSLRAEDTAVYYCAAGPLYTCVQSIVWPARPYYDMDYWGQ
		GTLVTVSS
87	CD8 VHHb	EVQLLESGGGLVQPGGSLRLSCAASGFTFDSYAIGWFRQAPG
	S75A/D31S	KEREGVSCIRVSDGSTYYADSVKGRFTISRDNAKNTLYLQM
		NSLRAEDTAVYYCAAGSLYTCVQSIVWPARPYYDMDYWGQ
		GTLVTVSS
88	CD8 VHHb	EVQLLESGGGLVQPGGSLRLSCAASGFTFPDYAIGWFRQAPG
	D30P/S75A/	KEREGVSCITVSDGSTYYADSVKGRFTISRDNAKNTLYLQM
	R52T	NSLRAEDTAVYYCAAGSLYTCVQSIVWPARPYYDMDYWGQ
		GTLVTVSS
89	CD8 VHHb	EVQLLESGGGLVQPGGSLRLSCAASGFTFPDYAIGWFRQAPG
	D30P/S75A	KEREGVSCIRVSDGSTYYADSVKGRFTISRDNAKNTLYLQM
		NSLRAEDTAVYYCAAGSLYTCVQSIVWPARPYYDMDYWGQ
		GTLVTVSS
90	IgG1 IMGT	EPKSCDK
	Upper Hinge
	Region
91	IMGT Lower	THTCPPCP
	Hinge Region
92	LM1486	EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYNMHWVRQA
	CD20 VH-	PGKGLEWVGAIYPGSGDTSYNQKFKGRFTISVDKSKNTLYL
	CH1 (IMGT)	QMNSLRAEDTAVYYCARVVYYSNSYWYFDVWGQGTLVTV
		SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS
		WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
		NVNHKPSNTKVDKRV
93	LM1653	QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYEMHWVRQ
	GPC3 VH-	APGQGLEWMGALDPKTGDTAYSQKFKGRVTLTADKSTSTA
	CH1 (IMGT)	YMELSSLTSEDTAVYYCTRFYSYTYWGQGTLVTVSSASTKG
		PSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT
		SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS
		NTKVDKRV
94	LRC150016	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	VH-CH1	PGQGLEWIGEILPGSDTTNYNEKFKDRATFTSDTSINTAYME
	(IMGT)	LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAS
		TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
		ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
		KPSNTKVDKRV
95	WT CH1	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
		NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
		NVNHKPSNTKVDKRVEPKSC
96	WT CH1	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
	(IMGT)	NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
		NVNHKPSNTKVDKRV
97	V12 CH1	ASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
	(F126C/C220V)	NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
		NVNHKPSNTKVDKRVEPKSV
98	WT CLλ	GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAW
		KADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHR
		SYSCQVTHEGSTVEKTVAPTECS
99	V12 LC	GQPKAAPSVTLFPPCSEELQANKATLVCLISDFYPGAVTVAW
	lambda	KADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHR
	constant	SYSCQVTHEGSTVEKTVAPTEVS
	(CLλ)
100	WT LC kappa	RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK
	constant (Cκ)	VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHK
	region	VYACEVTHQGLSSPVTKSFNRGEC
101	IgG1 CH2	ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
	Wild-type	NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
		NGKEYKCKVSNKALPAPIEKTIS
102	IgG1 CH2 Fc-	PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKF
	null	NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
		NGKEYKCKVSNKALPAPIEKTIS
103	IgG1 CH3	GQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWE
	Wild-type	SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF
		SCSVMHEALHNHYTQKSLSLSPGK
104	CH1 with	ASTKGPSVFPLAPSSKSTSGGTASLGCLVKDYFPEPVTVSW
	A141S	NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
		NVNHKPSNTKVDKRVEPKSC
105	CLλ with	GQPKAAPSVRLFPPSSEELQANKATLVCLISDFYPGAVTVAW
	T117R	KADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHR
		SYSCQVTHEGSTVEKTVAPTECS
106	V12 CH1 with	ASTKGPSVCPLAPSSKSTSGGTASLGCLVKDYFPEPVTVSW
	A141S	NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
		NVNHKPSNTKVDKRVEPKSV
107	V12 CLλ with	GQPKAAPSVRLFPPCSEELQANKATLVCLISDFYPGAVTVAW
	T117R	KADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHR
		SYSCQVTHEGSTVEKTVAPTEVS
108	IgG1 CH2	APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
	WT (IMGT)	KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
		WLNGKEYKCKVSNKALPAPIEKTIS
109	IgG1 CH2 Fc-	APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV
	null (IMGT)	KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
		WLNGKEYKCKVSNKALPAPIEKTIS
110	LM1486	EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYNMHWVRQA
	CD20 VH-	PGKGLEWVGAIYPGSGDTSYNQKFKGRFTISVDKSKNTLYL
	CH-TCR-	QMNSLRAEDTAVYYCARVVYYSNSYWYFDVWGQGTLVTV
	CH2CH3	SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS
	(hole/RF)	WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
		NVNHKPSNTKVDKRVEPKSCDKTGGSEVQLVESGGGLVQPG
		GSLRLSCVASGDVHKINFLGWYRQAPGKEREKVAHISIGDQT
		DYADSAKGRFTISRDESKNMVYLQMNSLKPEDTAVYFCRAF
		SRIYPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVF
		LFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGV
		EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
		VSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSL
		SCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV
		SKLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
111	TPP-42197	SYSMN
	BCMA
	VH CDR1
112	TPP-42197	SISGQSNYIYYADSVKG
	BCMA
	VH CDR2
113	TPP-42197	GGNYFVEYFQY
	BCMA
	VH CDR3
114	TPP-42197	RASQYISSNNLA
	BCMA VL
	CDR1
115	TPP-42197	GASNRAT
	BCMA VL
	CDR2
116	TPP-42197	QQYADSPIT
	BCMA VL
	CDR3
117	TPP-42197	EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAP
	BCMA VH	GKGLEWVSSISGQSNYIYYADSVKGRFTISRDNAKNSLYLQM
		NSLRAEDTAVYYCARGGNYFVEYFQYWGQGTLVTVSS
118	TPP-42197	EIVLTQSPGTLSLSPGERATLSCRASQYISSNNLAWYQQKPGQ
	BCMA VL	APRLLIYGASNRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVY
		YCQQYADSPITFGQGTKLEIK
119	TPP-42197	EIVLTQSPGTLSLSPGERATLSCRASQYISSNNLAWYQQKPGQ
	BCMA light	APRLLIYGASNRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVY
	chain	YCQQYADSPITFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGT
		ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSK
		DSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR
		GEC
120	TPP-42197	EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAP
	BCMA VH-	GKGLEWVSSISGQSNYIYYADSVKGRFTISRDNAKNSLYLQM
	CH1	NSLRAEDTAVYYCARGGNYFVEYFQYWGQGTLVTVSSAST
		KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
		LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK
		PSNTKVDKRVEPKSCDK
121	TPP-42197	EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAP
	hole (TCR)	GKGLEWVSSISGQSNYIYYADSVKGRFTISRDNAKNSLYLQM
	BCMA HC	NSLRAEDTAVYYCARGGNYFVEYFQYWGQGTLVTVSSAST
		KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
		LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK
		PSNTKVDKRVEPKSCDKTGGSEVQLVESGGGLVQPGGSLRL
		SCVASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTDYAD
		SAKGRFTISRDESKNMVYLQMNSLKPEDTAVYFCRAFSRIYP
		YDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPPK
		PKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVHN
		AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
		ALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCAV
		KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLT
		VDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
122	TPP-42197-	EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAP
	knob (CD8)	GKGLEWVSSISGQSNYIYYADSVKGRFTISRDNAKNSLYLQM
	BCMA HC	NSLRAEDTAVYYCARGGNYFVEYFQYWGQGTLVTVSSAST
		KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
		LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK
		PSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGSLRLS
		CAASGFTFDDYAIGWFRQAPGKEREGVSCIRVSDGSTYYADS
		VKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAAGSLYTC
		VQSIVWPARPYYDMDYWGQGTLVTVSSGGGGSTHTCPPCP
		APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV
		KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
		WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPC
		REEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTP
		PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
		TQKSLSLSPGK
123	LM1953	RNSAVWN
	STEAP2 HC
	CDR1
124	LM1953	RTYYRSKWYNDYAVSVKS
	STEAP2 HC
	CDR2
125	LM1953	GLRQNQFYYYMDV
	STEAP2 HC
	CDR3
126	LM1953	RASQSVASNLA
	STEAP2 LC
	CDR1
127	LM1953	GASTRAT
	STEAP2 LC
	CDR2
128	LM1953	QQYNNWPFT
	STEAP2 LC
	CDR3
129	LM1953	QVQLQQSGPGLVKPSQTLSLTCAISGDSVSRNSAVWNWIRQS
	STEAP2 VH	PSRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSL
		QLNSVTPEDTAVYYCARGLRQNQFYYYMDVWGKGTTVTVS
		S
130	LM1953	EIVMTQSPATLSVSPGERATLSCRASQSVASNLAWYQQKPG
	STEAP2 VL	QAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAV
		YYCQQYNNWPFTFGPGTKVDIK
131	LM1953	EIVMTQSPATLSVSPGERATLSCRASQSVASNLAWYQQKPG
	STEAP2 LC	QAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAV
		YYCQQYNNWPFTFGPGTKVDIKRTVAAPSVFIFPPSDEQLKS
		GTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD
		SKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSF
		NRGEC
132	LM1953	QVQLQQSGPGLVKPSQTLSLTCAISGDSVSRNSAVWNWIRQS
	STEAP2 VH-	PSRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSL
	CH1	QLNSVTPEDTAVYYCARGLRQNQFYYYMDVWGKGTTVTVS
		SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
		NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN
		VNHKPSNTKVDKRVEPKSCDK
133	LM1953	QVQLQQSGPGLVKPSQTLSLTCAISGDSVSRNSAVWNWIRQS
	STEAP2 knob	PSRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSL
	(CD8)	QLNSVTPEDTAVYYCARGLRQNQFYYYMDVWGKGTTVTVS
		SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
		NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN
		VNHKPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGG
		SLRLSCAASGFTFDDYAIGWFRQAPGKEREGVSCIRVSDGST
		YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAAG
		SLYTCVQSIVWPARPYYDMDYWGQGTLVTVSSGGGGSTHT
		CPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHE
		DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL
		HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT
		LPPCREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNY
		KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL
		HNHYTQKSLSLSPGK
134	LM1953	QVQLQQSGPGLVKPSQTLSLTCAISGDSVSRNSAVWNWIRQS
	STEAP2 hole	PSRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSL
	(TCR)	QLNSVTPEDTAVYYCARGLRQNQFYYYMDVWGKGTTVTVS
		SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
		NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN
		VNHKPSNTKVDKRVEPKSCDKTGGSEVQLVESGGGLVQPGG
		SLRLSCVASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTD
		YADSAKGRFTISRDESKNMVYLQMNSLKPEDTAVYFCRAFS
		RIYPYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFL
		FPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVE
		VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
		SNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLS
		CAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVS
		KLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
135	TPP-46956	SYWIE
	TPP-46957
	TPP-46958
	TPP-46959
	TPP-46960
	TPP-47826
	TPP-49058
	LRRC15 VH
	CDR1
136	TPP-46956	EILPGSDTTNYAQNFQD
	LRRC15 VH
	CDR2
137	TPP-46956	DRGNYRAWFGY
	TPP-46957
	TPP-46958
	TPP-46959
	TPP-46960
	TPP-47826
	TPP-49058
	LRRC15 VH
	CDR3
138	TPP-46956	RASQDISNYLN
	TPP-46957
	TPP-46958
	TPP-46959
	TPP-46960
	TPP-47826
	TPP-49058
	LRRC15 VL
	CDR1
139	TPP-46956	YTSRLHS
	TPP-46957
	TPP-46958
	TPP-47826
	LRRC15 VL
	CDR2
140	TPP-46956	QQGNALPWT
	TPP-46957
	TPP-46958
	TPP-46959
	TPP-46960
	TPP-47826
	TPP-49058
	LRRC15 VL
	CDR3
141	TPP-46956	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	LRRC15 VH	PGQGLEWIGEILPGSDTTNYAQNFQDRVTFTSDTSISTAYMEL
		SRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSS
142	TPP-46956	DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQKPGK
	TPP-46957	APKFLIYYTSRLHSGVPSRFSGSGSGTDYTLTISSLQPEDFATY
	TPP-46958	YCQQGNALPWTFGGGTKVEIK
	TPP-47826
	LRRC15 VL
143	TPP-46956	DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQKPGK
	TPP-46957	APKFLIYYTSRLHSGVPSRFSGSGSGTDYTLTISSLQPEDFATY
	TPP-46958	YCQQGNALPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSG
	TPP-47826	TASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS
	LRRC15	KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFN
	Light chain	RGEC
144	TPP-46956	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	LRRC15 VH-	PGQGLEWIGEILPGSDTTNYAQNFQDRVTFTSDTSISTAYMEL
	CH1	SRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAST
		KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
		LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK
		PSNTKVDKRVEPKSCDK
145	TPP-46956	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	LRRC15 hole	PGQGLEWIGEILPGSDTTNYAQNFQDRVTFTSDTSISTAYMEL
	(TCR)	SRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAST
		KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
		LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK
		PSNTKVDKRVEPKSCDKTGGSEVQLVESGGGLVQPGGSLRL
		SCVASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTDYAD
		SAKGRFTISRDESKNMVYLQMNSLKPEDTAVYFCRAFSRIYP
		YDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPPK
		PKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVHN
		AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
		ALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCAV
		KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLT
		VDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
146	TPP-46956	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	LRRC15	PGQGLEWIGEILPGSDTTNYAQNFQDRVTFTSDTSISTAYMEL
	knob (CD8)	SRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAST
		KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
		LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK
		PSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGSLRLS
		CAASGFTFTDYAIGWFRQAPGKEREGVSCIRVSDGSTYYADS
		VKGRFTISRDSSKNTLYLQMNSLRAEDTAVYYCAAGSLYTC
		VQSIVWPARPYYDMDYWGQGTLVTVSSGGGGSTHTCPPCP
		APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV
		KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
		WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPC
		REEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTP
		PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
		TQKSLSLSPGK
147	TPP-46957	EILPGSDTTNYAQQFQD
	LRRC15 VH
	CDR2
148	TPP-46957	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	LRRC15 VH	PGQGLEWIGEILPGSDTTNYAQQFQDRVTFTSDTSISTAYMEL
		SRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSS
149	TPP-46957	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	LRRC15 VH-	PGQGLEWIGEILPGSDTTNYAQQFQDRVTFTSDTSISTAYMEL
	CH1	SRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAST
		KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
		LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK
		PSNTKVDKRVEPKSCDK
150	TPP-46957	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	LRRC15 hole	PGQGLEWIGEILPGSDTTNYAQQFQDRVTFTSDTSISTAYMEL
	(TCR)	SRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAST
		KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
		LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK
		PSNTKVDKRVEPKSCDKTGGSEVQLVESGGGLVQPGGSLRL
		SCVASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTDYAD
		SAKGRFTISRDESKNMVYLQMNSLKPEDTAVYFCRAFSRIYP
		YDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPPK
		PKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVHN
		AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
		ALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCAV
		KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLT
		VDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
151	TPP-46957	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	LRRC15	PGQGLEWIGEILPGSDTTNYAQQFQDRVTFTSDTSISTAYMEL
	knob (CD8)	SRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAST
		KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
		LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK
		PSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGSLRLS
		CAASGFTFTDYAIGWFRQAPGKEREGVSCIRVSDGSTYYADS
		VKGRFTISRDSSKNTLYLQMNSLRAEDTAVYYCAAGSLYTC
		VQSIVWPARPYYDMDYWGQGTLVTVSSGGGGSTHTCPPCP
		APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV
		KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
		WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPC
		REEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTP
		PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
		TQKSLSLSPGK
152	TPP-46958	EILPGSDATNYAQKFQD
	TPP-47826
	LRRC15 VH
	CDR2
153	TPP-46958	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	TPP-47826	PGQGLEWIGEILPGSDATNYAQKFQDRVTFTSDTSISTAYME
	LRRC15 VH	LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSS
154	TPP-46958	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	TPP-47826	PGQGLEWIGEILPGSDATNYAQKFQDRVTFTSDTSISTAYME
	LRRC15 VH-	LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAS
	CH1	TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
		ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
		KPSNTKVDKRVEPKSCDK
155	TPP-46958	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	LRRC15 hole	PGQGLEWIGEILPGSDATNYAQKFQDRVTFTSDTSISTAYME
	(TCR)	LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAS
		TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
		ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
		KPSNTKVDKRVEPKSCDKTGGSEVQLVESGGGLVQPGGSLR
		LSCVASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTDYA
		DSAKGRFTISRDESKNMVYLQMNSLKPEDTAVYFCRAFSRIY
		PYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPP
		KPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVH
		NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
		KALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCA
		VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKL
		TVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
156	TPP-46958	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	TPP-47826	PGQGLEWIGEILPGSDATNYAQKFQDRVTFTSDTSISTAYME
	LRRC15	LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAS
	knob (CD8)	TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
		ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
		KPSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGSLR
		LSCAASGFTFTDYAIGWFRQAPGKEREGVSCIRVSDGSTYYA
		DSVKGRFTISRDSSKNTLYLQMNSLRAEDTAVYYCAAGSLY
		TCVQSIVWPARPYYDMDYWGQGTLVTVSSGGGGSTHTCPP
		CPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDP
		EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ
		DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPP
		CREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT
		PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
		YTQKSLSLSPGK
157	TPP-46959	EILPGSDTTNYAQKFQD
	TPP-46960
	TPP-49058
	LRRC15 VH
	CDR2
158	TPP-46959	YTSRLES
	TPP-49058
	LRRC15 VL
	CDR2
159	TPP-46959	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	TPP-46960	PGQGLEWIGEILPGSDTTNYAQKFQDRVTFTSDTSISTAYMEL
	TPP-49058	SRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSS
	LRRC15 VH
160	TPP-46959	DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQKPGK
	TPP-49058	APKFLIYYTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATY
	LRRC15 VL	YCQQGNALPWTFGGGTKVEIK
161	TPP-46959	DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQKPGK
	TPP-49058	APKFLIYYTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATY
	LRRC15	YCQQGNALPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSG
	Light chain	TASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS
		KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFN
		RGEC
162	TPP-46959	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	TPP-46960	PGQGLEWIGEILPGSDTTNYAQKFQDRVTFTSDTSISTAYMEL
	TPP-49058	SRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAST
	LRRC15 VH-	KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
	CH1	LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK
		PSNTKVDKRVEPKSCDK
163	TPP-46959	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	TPP-46960	PGQGLEWIGEILPGSDTTNYAQKFQDRVTFTSDTSISTAYMEL
	LRRC15 hole	SRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAST
	(TCR)	KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
		LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK
		PSNTKVDKRVEPKSCDKTGGSEVQLVESGGGLVQPGGSLRL
		SCVASGDVHKINFLGWYRQAPGKEREKVAHISIGDQTDYAD
		SAKGRFTISRDESKNMVYLQMNSLKPEDTAVYFCRAFSRTYP
		YDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPPK
		PKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVHN
		AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
		ALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCAV
		KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLT
		VDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
164	TPP-46959	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	TPP-46960	PGQGLEWIGEILPGSDTTNYAQKFQDRVTFTSDTSISTAYMEL
	TPP-49058	SRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAST
	LRRC15	KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
	knob (CD8)	LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK
		PSNTKVDKRVEPKSCDKTGGSEVQLLESGGGLVQPGGSLRLS
		CAASGFTFTDYAIGWFRQAPGKEREGVSCIRVSDGSTYYADS
		VKGRFTISRDSSKNTLYLQMNSLRAEDTAVYYCAAGSLYTC
		VQSIVWPARPYYDMDYWGQGTLVTVSSGGGGSTHTCPPCP
		APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV
		KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
		WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPC
		REEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTP
		PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
		TQKSLSLSPGK
165	TPP-46960	YTSRLNS
	LRRC15 VL
	CDR2
166	TPP-46960	DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQKPGK
	LRRC15 VL	APKFLIYYTSRLNSGVPSRFSGSGSGTDYTLTISSLQPEDFATY
		YCQQGNALPWTFGGGTKVEIK
167	TPP-46960	DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQKPGK
	LRRC15	APKFLIYYTSRLNSGVPSRFSGSGSGTDYTLTISSLQPEDFATY
	Light chain	YCQQGNALPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSG
		TASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS
		KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFN
		RGEC
168	CD8 VHHb	EVQLLESGGGLVQPGGSLRLSCAASGFTFTDYAIGWFRQAPG
	D30T/N74S	KEREGVSCIRVSDGSTYYADSVKGRFTISRDSSKNTLYLQMN
		SLRAEDTAVYYCAAGSLYTCVQSIVWPARPYYDMDYWGQG
		TLVTVSS
169	CD8 VHHb	GFTFTDYA
	D30T/N74S
	CDR1
	IMGT
170	CD8 VHHb	IRVSDGST
	D30T/N74S
	CDR2
	IMGT
171	CD8 VHHb	AAGSLYTCVQSIVWPARPYYDMDY
	D30T/N74S
	CDR3
	IMGT
172	TCR VHHa	DVQLVESGGGVVQPGGSLRLSCVASGYVHKINFYGWYRQA
	alternative	PGKEREKVAHISIGDQTDYADSAKGRFTISRDESKNTVYLQM
		NSLRPEDTAAYYCRALSRIWPYDYWGQGTLVTVSS
173	TCR VHHa	INFYG
	CDR1
	alternative
174	TCR VHHa	HISIGDQTDYADSAKG
	CDR2
	alternative
175	TCR VHHa	LSRIWPYDY
	CDR3
	alternative
176	TPP-47826	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	LRRC15 hole	PGQGLEWIGEILPGSDATNYAQKFQDRVTFTSDTSISTAYME
	(TCR)	LSRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAS
		TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
		ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH
		KPSNTKVDKRVEPKSCDKTGGSDVQLVESGGGVVQPGGSLR
		LSCVASGYVHKINFYGWYRQAPGKEREKVAHISIGDQTDYA
		DSAKGRFTISRDESKNTVYLQMNSLRPEDTAAYYCRALSRIW
		PYDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPP
		KPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVH
		NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
		KALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCA
		VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKL
		TVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
177	TPP-49058	EVQLVQSGAEVKKPGASVKVSCKASGYKFSSYWIEWVKQA
	LRRC15 hole	PGQGLEWIGEILPGSDTTNYAQKFQDRVTFTSDTSISTAYMEL
	(TCR)	SRLRSDDTAVYYCARDRGNYRAWFGYWGQGTLVTVSSAST
		KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
		LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK
		PSNTKVDKRVEPKSCDKTGGSDVQLVESGGGVVQPGGSLRL
		SCVASGYVHKINFYGWYRQAPGKEREKVAHISIGDQTDYAD
		SAKGRFTISRDESKNTVYLQMNSLRPEDTAAYYCRALSRIWP
		YDYWGQGTLVTVSSGGGGSTHTCPPCPAPPVAGPSVFLFPPK
		PKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVHN
		AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
		ALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLSCAV
		KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLT
		VDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK
178	Comparator T	EVQLVESGGGLVQPGGSLRLSCAASGFTFNTYAMNWVRQA
	HC (CD3)	PGKGLEWVARIRSKYNNYATYYAASVKGRFTISRDDSKNSL
		YLQMNSLKTEDTAVYYCARHGNFGNSYVSWFAYWGQGTL
		VTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVT
		VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKT
		YTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFL
		FPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE
		VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKV
		SNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLT
		CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLLYS
		KLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK
179	Comparator T	QTVVTQEPSLTVSPGGTVTLTCRSSTGAVTTSNYANWVQQK
	LC (CD3)	PGQAPRGLIGGTNKRAPGTPARFSGSLLGGKAALTLSGVQPE
		DEAEYYCALWYSNLWVFGGGTKLTVLGQPKAAPSVTLFPPS
		SEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETT
		TPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVE
		KTVAPTECS
180	Comparator T	QLQLQESGPGLVKPSETLSLTCTVSGGSISSGSYFWGWIRQPP
	HC (BCMA)	GKGLEWIGSIYYSGITYYNPSLKSRVTISVDTSKNQFSLKLSS
		VTAADTAVYYCARHDGAVAGLFDYWGQGTLVTVSSASTKG
		PSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTS
		GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPS
		NTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLM
		ISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPRE
		EQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEK
		TISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDI
		AVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQ
		EGNVFSCSVMHEALHNHYTQKSLSLSLGK
181	Comparator T	SYVLTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQPPGQA
	LC (BCMA)	PVVVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAV
		YYCQVWDSSSDHVVFGGGTKLTVLGQPKAAPSVTLFPPSSE
		ELQANKATLVCLISDFYPGAVTVAWKGDSSPVKAGVETTTP
		SKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKT
		VAPTECS
182	LM1486/LM1486-	SYNMH
	2 CD20
	VH CDR1
	Kabat
183	CD8 VHHb	CIRVSDGSTYYADSVKG
	CDR2
	(alternate)
	LM1653
184	GPC3	CAAGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAA
	Heavy Chain	CCTGGCGCCTCTGTGAAGGTGTCCTGCAAGGCTTCTGGCT
	Hole (TCR)	ACACCTTTACCGACTACGAGATGCACTGGGTCCGACAGGC
		TCCAGGACAAGGCTTGGAATGGATGGGCGCCCTGGATCCT
		AAGACCGGCGATACCGCTTACTCCCAGAAATTCAAGGGCA
		GAGTGACCCTGACCGCCGACAAGTCTACCTCCACCGCCTA
		CATGGAACTGTCCAGCCTGACCTCTGAGGACACCGCCGTG
		TACTACTGCACCCGGTTCTACTCCTACACCTACTGGGGCC
		AGGGCACCCTGGTTACAGTGTCCTCTGCTTCCACCAAGGG
		ACCCAGCGTTTTCCCTCTGGCTCCATCCTCCAAGTCCACCT
		CTGGTGGAACAGCTGCTCTGGGCTGCCTGGTCAAGGACTA
		CTTTCCTGAGCCTGTGACCGTGTCCTGGAACTCTGGCGCTC
		TGACATCTGGCGTGCACACCTTTCCAGCTGTGCTGCAGTC
		CTCCGGCCTGTACTCTCTGTCCTCTGTCGTGACCGTGCCTT
		CCAGCTCTCTGGGAACCCAGACCTACATCTGCAATGTGAA
		CCACAAGCCTTCCAATACCAAGGTGGACAAGAGAGTGGA
		ACCCAAGTCCTGCGACAAGACAGGCGGATCTGAGGTCCA
		GCTGGTCGAATCTGGCGGAGGATTGGTTCAGCCTGGCGGC
		TCTCTGAGACTGTCTTGTGTGGCTAGCGGCGACGTGCACA
		AGATCAACTTTCTCGGCTGGTACAGACAGGCCCCTGGCAA
		AGAGAGAGAGAAGGTCGCCCACATCTCCATCGGCGACCA
		GACCGATTACGCCGACTCTGCTAAGGGCAGATTCACCATC
		TCTCGGGACGAGTCCAAGAACATGGTGTACCTGCAGATGA
		ACTCCCTGAAGCCAGAGGATACCGCTGTGTATTTCTGCCG
		GGCCTTCAGCCGGATCTACCCTTACGATTATTGGGGACAG
		GGAACACTCGTGACAGTGTCTAGCGGAGGCGGAGGCTCT
		ACCCATACCTGTCCTCCATGTCCTGCTCCTCCAGTGGCTGG
		CCCTTCCGTGTTTCTGTTCCCTCCAAAGCCTAAGGACACCC
		TGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGT
		GGATGTGAAGCACGAGGACCCAGAAGTGAAGTTCAATTG
		GTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAA
		GCCTAGAGAGGAACAGTACAACTCCACCTACAGAGTGGT
		GTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGC
		AAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGCCT
		GCTCCTATCGAAAAGACCATCTCCAAGGCCAAGGGACAG
		CCCCGGGAACCTCAAGTCTGTACCCTGCCTCCTAGCCGGG
		AAGAGATGACCAAGAACCAGGTGTCCCTGTCCTGTGCCGT
		GAAGGGCTTCTACCCCTCTGATATCGCCGTGGAATGGGAG
		TCCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTC
		CTGTGCTGGACTCCGACGGCTCATTCTTCCTGGTGTCCAAG
		CTGACAGTGGACAAGTCCAGATGGCAGCAGGGCAACGTG
		TTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAATCACT
		ACACACAGAAGTCCCTGTCTCTGAGCCCCGGCAAA
185	LM1653	CAAGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAA
	GPC3	CCTGGCGCCTCTGTGAAGGTGTCCTGCAAGGCTTCTGGCT
	Heavy Chain	ACACCTTTACCGACTACGAGATGCACTGGGTCCGACAGGC
	Knob (CD8)	TCCAGGACAAGGCTTGGAATGGATGGGCGCCCTGGATCCT
		AAGACCGGCGATACCGCTTACTCCCAGAAATTCAAGGGCA
		GAGTGACCCTGACCGCCGACAAGTCTACCTCCACCGCCTA
		CATGGAACTGTCCAGCCTGACCTCTGAGGACACCGCCGTG
		TACTACTGCACCCGGTTCTACTCCTACACCTACTGGGGCC
		AGGGCACCCTGGTTACAGTGTCCTCTGCTTCCACCAAGGG
		ACCCAGCGTTTTCCCTCTGGCTCCATCCTCCAAGTCCACCT
		CTGGTGGAACAGCTGCTCTGGGCTGCCTGGTCAAGGACTA
		CTTTCCTGAGCCTGTGACCGTGTCCTGGAACTCTGGCGCTC
		TGACATCTGGCGTGCACACCTTTCCAGCTGTGCTGCAGTC
		CTCCGGCCTGTACTCTCTGTCCTCTGTCGTGACCGTGCCTT
		CCAGCTCTCTGGGAACCCAGACCTACATCTGCAATGTGAA
		CCACAAGCCTTCCAACACCAAGGTCGACAAGAGAGTGGA
		ACCCAAGTCCTGCGACAAGACAGGCGGATCTGAGGTGCA
		GCTGCTCGAATCTGGCGGAGGATTGGTTCAGCCTGGCGGC
		TCTCTGAGACTGTCTTGTGCTGCCTCCGGCTTCACCTTCGA
		CGATTACGCCATCGGCTGGTTCAGACAGGCCCCTGGCAAA
		GAGAGAGAGGGCGTCAGCTGCATCAGAGTGTCTGACGGC
		TCTACCTACTACGCCGACTCCGTGAAGGGCAGATTCACCA
		TCTCTCGGGACAACTCCAAGAACACCCTGTACCTGCAGAT
		GAACTCCCTGAGAGCCGAGGATACCGCTGTGTATTATTGC
		GCCGCTGGCTCCCTGTATACCTGCGTGCAGTCTATCGTGTG
		GCCCGCCAGACCTTACTACGACATGGACTATTGGGGACAG
		GGAACACTCGTGACAGTGTCTAGCGGAGGCGGCGGATCT
		ACCCATACCTGTCCTCCATGTCCTGCTCCTCCAGTGGCTGG
		CCCTTCCGTGTTTCTGTTCCCTCCAAAGCCTAAGGACACCC
		TGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGT
		GGATGTGAAGCACGAGGACCCAGAAGTGAAGTTCAATTG
		GTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAA
		GCCTAGAGAGGAACAGTACAACTCCACCTACAGAGTGGT
		GTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGC
		AAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGCCT
		GCTCCTATCGAAAAGACCATCTCCAAGGCCAAGGGACAG
		CCCAGGGAACCCCAGGTTTACACCCTGCCTCCATGCCGGG
		AAGAGATGACCAAGAACCAGGTGTCCCTGTGGTGCCTGGT
		TAAGGGCTTCTACCCCTCCGATATCGCCGTGGAATGGGAG
		TCTAATGGCCAGCCTGAGAACAACTACAAGACCACACCTC
		CTGTGCTGGACTCCGACGGCTCATTCTTCCTGTACTCCAAG
		CTGACAGTGGACAAGTCCAGATGGCAGCAGGGCAACGTG
		TTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAATCACT
		ACACACAGAAGTCCCTGTCTCTGAGCCCCGGCAAA
186	LM1653	GACGTCGTGATGACCCAGTCTCCTCTGTCTCTGCCTGTGAC
	GPC3	ACCTGGCGAGCCTGCCTCCATCTCTTGCAGATCTTCTCAGT
	Light Chain	CCCTGGTGCACTCCAACCGGAACACCTACCTGCACTGGTA
		TCTGCAGAAGCCCGGACAGTCTCCCCAGCTGCTGATCTAC
		AAGGTGTCCAACAGATTCTCTGGCGTGCCCGACAGATTCA
		GCGGCTCTGGCTCTGGCACCGACTTCACCCTGAAGATCTC
		TAGAGTGGAAGCCGAGGACGTGGGCGTGTACTACTGCTCT
		CAGAATACCCACGTGCCACCTACCTTTGGCCAGGGCACCA
		AGCTGGAAATCAAGAGAACCGTGGCCGCTCCTTCCGTGTT
		CATCTTCCCACCATCTGACGAGCAGCTGAAGTCCGGCACA
		GCTTCTGTCGTGTGCCTGCTGAACAACTTCTACCCTCGGGA
		AGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGTC
		CGGCAACTCCCAAGAGTCTGTGACCGAGCAGGACTCCAA
		GGACTCTACCTACAGCCTGTCCTCCACACTGACCCTGTCTA
		AGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAAG
		TGACCCACCAGGGCCTGTCTAGCCCTGTGACCAAGTCTTT
		CAACCGGGGCGAGTGC
187	LM1486-2	GAGGTGCAGCTGGTTGAATCTGGCGGAGGATTGGTTCAGC
	CD20	CTGGCGGCTCTCTGAGACTGTCTTGTGCTGCCTCTGGCTAC
	Heavy Chain	ACCTTCACCAGCTACAACATGCACTGGGTCCGACAGGCCC
	Hole (TCR)	CTGGCAAAGGATTGGAATGGGTCGGAGCTATCTACCCTGG
		CTCCGGCGATACCTCCTACAACCAGAAGTTCAAGGGCAGA
		TTCACCATCTCCGTGGACAAGTCCAAGAACACCCTGTACC
		TGCAGATGAACTCCCTGAGAGCCGAGGACACCGCCGTGTA
		CTATTGTGCCAGAGTGGTGTACTACTCCAACTCCTACTGGT
		ACTTCGACGTGTGGGGCCAGGGAACACTGGTCACAGTGTC
		CTCTGCTTCCACCAAGGGACCCTCTGTGTTCCCTCTGGCTC
		CTTCCAGCAAGTCTACCTCTGGTGGAACCGCTGCTCTGGG
		CTGCCTGGTCAAGGATTACTTTCCTGAGCCTGTGACCGTGT
		CCTGGAATTCTGGTGCTCTGACCTCCGGCGTGCACACCTTT
		CCAGCTGTGCTGCAGTCCTCTGGCCTGTACTCTCTGTCCTC
		TGTCGTGACCGTGCCTTCTAGCTCTCTGGGCACCCAGACCT
		ACATCTGCAACGTGAACCACAAGCCTTCCAACACCAAGGT
		GGACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACCGG
		CGGATCTGAAGTCCAGTTGGTGGAAAGCGGAGGTGGACTT
		GTGCAGCCAGGTGGAAGCCTGAGACTGAGTTGTGTGGCTT
		CTGGCGACGTGCACAAGATCAACTTTCTCGGCTGGTACAG
		GCAGGCTCCCGGCAAAGAAAGGGAAAAAGTGGCCCACAT
		CTCCATCGGCGACCAGACCGATTACGCCGACTCTGCCAAA
		GGCCGGTTTACCATCTCTCGGGACGAGAGCAAGAACATGG
		TGTATCTCCAGATGAACAGTCTGAAGCCCGAGGATACAGC
		TGTGTACTTCTGCCGGGCCTTCAGCCGGATCTACCCTTACG
		ATTATTGGGGACAGGGCACCCTGGTTACCGTTTCTAGTGG
		CGGAGGCGGCTCTACCCATACCTGTCCTCCATGTCCTGCTC
		CACCTGTGGCCGGCCCTTCCGTGTTCCTGTTTCCTCCAAAG
		CCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAGTGA
		CCTGCGTGGTGGTGGATGTGAAACACGAGGATCCCGAAGT
		GAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAA
		CGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCAC
		CTACCGCGTGGTGTCTGTGCTGACCGTTCTGCACCAGGAC
		TGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAAC
		AAGGCCCTGCCTGCCCCCATCGAAAAGACCATCTCTAAGG
		CTAAGGGCCAGCCTCGCGAGCCTCAAGTCTGTACACTGCC
		TCCTAGCCGGGAAGAGATGACCAAGAATCAGGTGTCCCTG
		TCCTGCGCCGTGAAGGGCTTCTACCCTTCTGATATCGCCGT
		GGAATGGGAGTCCAACGGCCAGCCTGAGAACAACTACAA
		GACCACACCTCCTGTGCTGGACTCCGACGGCTCATTCTTCC
		TGGTGTCCAAGCTGACCGTGGATAAGAGCAGATGGCAGC
		AGGGCAACGTGTTCTCCTGCTCTGTGATGCACGAGGCCCT
		GCACAACCACTATACCCAGAAATCCCTGTCTCTGAGCCCT
		GGCAAG
188	LM1486/	GAGGTGCAGCTGGTGGAATCTGGCGGAGGATTGGTTCAGC
	LM1486-2	CTGGCGGCTCTCTGAGACTGTCTTGTGCTGCCTCCGGCTAC
	CD20	ACCTTCACCAGCTACAACATGCACTGGGTCCGACAGGCCC
	Heavy Chain	CTGGCAAAGGATTGGAATGGGTCGGAGCTATCTACCCCGG
	Knob (CD8)	CTCTGGCGACACCTCCTACAACCAGAAGTTCAAGGGCAGA
		TTCACCATCTCCGTGGACAAGTCCAAGAACACCCTGTACC
		TGCAGATGAACTCCCTGAGAGCCGAGGACACCGCCGTGTA
		CTACTGTGCCAGAGTGGTGTACTACAGCAACTCCTACTGG
		TACTTCGACGTGTGGGGCCAGGGCACACTGGTCACAGTTT
		CTTCCGCCTCCACCAAGGGACCCAGCGTTTTCCCTCTGGCT
		CCATCCTCCAAGTCTACCTCTGGCGGAACAGCTGCTCTGG
		GCTGCCTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGT
		GTCCTGGAACTCTGGCGCTCTGACATCTGGCGTGCACACC
		TTTCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTC
		CTCTGTCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAG
		ACCTACATCTGCAATGTGAACCACAAGCCTTCCAACACCA
		AGGTGGACAAGAGAGTGGAACCCAAGTCCTGCGATAAGA
		CCGGCGGAAGCGAAGTGCAGCTGCTTGAAAGCGGAGGTG
		GACTTGTGCAGCCAGGCGGAAGCCTGAGATTGTCCTGTGC
		CGCTTCTGGCTTTACCTTCGACGACTACGCCATCGGCTGGT
		TCAGACAGGCTCCCGGAAAAGAGAGAGAGGGCGTCAGCT
		GCATCAGAGTGTCTGACGGCTCTACCTACTACGCCGACTC
		CGTGAAAGGCCGGTTCACCATCAGCCGGGACAACAGCAA
		GAATACTCTGTATCTCCAAATGAACAGCCTGCGCGCTGAG
		GATACCGCTGTGTATTATTGCGCCGCTGGCTCCCTGTATAC
		CTGCGTGCAGTCTATCGTGTGGCCCGCCAGACCTTACTAC
		GACATGGATTACTGGGGACAGGGAACCCTGGTTACCGTGT
		CTAGCGGCGGAGGCGGATCTACCCATACCTGTCCTCCATG
		TCCTGCTCCACCTGTGGCCGGCCCTTCCGTGTTCCTGTTTC
		CTCCAAAGCCTAAGGACACCCTGATGATCTCTCGGACCCC
		TGAAGTGACCTGCGTGGTGGTGGATGTGAAACACGAGGA
		TCCCGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAA
		GTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTAC
		AACTCCACCTACCGCGTGGTGTCCGTGCTGACCGTTCTGC
		ATCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGG
		TGTCCAACAAGGCCCTGCCTGCCCCCATCGAAAAGACCAT
		CTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTAC
		ACCCTGCCACCTTGCCGGGAAGAGATGACCAAGAACCAG
		GTGTCCCTGTGGTGCCTCGTGAAGGGCTTCTACCCTTCCGA
		TATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAA
		CAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGC
		TCATTCTTCCTGTACTCCAAGCTGACCGTGGATAAGTCTCG
		GTGGCAGCAGGGCAACGTGTTCTCCTGCTCTGTGATGCAC
		GAGGCCCTGCACAACCACTATACCCAGAAGTCCCTGTCTC
		TGAGCCCCGGCAAG
189	LM1486/	GACATCCAGATGACCCAGTCTCCATCCTCTCTGTCTGCCTC
	LM1486-2	TGTGGGCGACAGAGTGACAATCACCTGTAGAGCCTCCAGC
	CD20	TCCGTGTCCTACATGCACTGGTATCAGCAGAAGCCCGGCA
	Light Chain	AGGCCCCTAAGCCTCTGATCTACGCTCCTTCTAATCTGGCC
		TCCGGCGTGCCCTCTAGATTTTCTGGCTCTGGATCTGGCAC
		CGACTTCACCCTGACCATCAGTTCTCTGCAGCCTGAGGAC
		TTCGCCACCTACTACTGCCAGCAGTGGTCTTTCAACCCTCC
		TACCTTTGGCCAGGGCACCAAGGTGGAAATCAAGAGAAC
		CGTGGCCGCTCCTTCCGTGTTCATCTTCCCACCATCTGACG
		AGCAGCTGAAGTCCGGCACAGCTTCTGTCGTGTGCCTGCT
		GAACAACTTCTACCCTCGGGAAGCCAAGGTGCAGTGGAA
		GGTGGACAATGCCCTGCAGTCCGGCAACTCCCAAGAGTCT
		GTGACCGAGCAGGACTCCAAGGACTCTACCTACAGCCTGT
		CCTCCACACTGACCCTGTCTAAGGCCGACTACGAGAAGCA
		CAAGGTGTACGCCTGTGAAGTGACCCACCAGGGACTGTCT
		AGCCCCGTGACCAAGTCCTTCAACAGAGGCGAGTGT
190	LM1953	CAAGTGCAGCTGCAGCAGTCAGGTCCAGGACTGGTGAAG
	STEAP2	CCCTCGCAGACCCTCTCACTCACCTGTGCCATCTCCGGGG
	Heavy Chain	ACAGTGTCTCCAGAAACAGTGCTGTTTGGAATTGGATCAG
	Hole (TCR)	GCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGAAGGAC
		ATACTACAGGTCCAAGTGGTATAATGATTATGCAGTATCT
		GTGAAAAGTCGAATAACCATCAACCCAGACACATCCAAG
		AACCAGTTCTCCCTGCAACTGAACTCTGTGACTCCCGAGG
		ACACGGCTGTGTATTACTGTGCAAGGGGGTTACGACAGAA
		CCAGTTCTACTACTACATGGACGTCTGGGGCAAAGGGACC
		ACGGTCACCGTCTCCTCCGCTAGTACCAAGGGACCCAGCG
		TGTTCCCTCTGGCACCTTCCAGCAAGTCTACCTCTGGCGGA
		ACAGCCGCTCTGGGCTGTCTGGTCAAGGACTACTTTCCCG
		AGCCTGTGACCGTGTCCTGGAATTCTGGCGCTCTGACCAG
		CGGAGTGCATACCTTTCCAGCTGTGCTGCAGTCCTCCGGC
		CTGTACTCTCTGTCCTCTGTCGTGACCGTGCCTTCCAGCTC
		TCTGGGCACCCAGACCTACATCTGCAATGTGAACCACAAG
		CCTTCCAACACCAAGGTGGACAAGAGGGTGGAACCCAAG
		TCCTGCGACAAGACCGGCGGATCTGAAGTGCAGCTGGTCG
		AGTCTGGCGGAGGATTGGTTCAACCTGGCGGCTCCCTGAG
		ACTGTCTTGTGTGGCTAGCGGAGATGTGCATAAGATCAAT
		TTTCTCGGCTGGTACAGACAGGCCCCTGGCAAAGAGAGAG
		AGAAGGTCGCCCACATCTCCATCGGCGACCAGACCGATTA
		CGCCGACTCTGCCAAGGGCAGATTCACCATCTCTCGGGAC
		GAGTCCAAGAACATGGTGTACCTGCAGATGAACTCCCTGA
		AGCCTGAGGATACCGCCGTGTACTTCTGCCGGGCCTTCTC
		TCGGATCTACCCCTACGATTATTGGGGCCAGGGCACCCTG
		GTCACAGTGTCATCTGGAGGCGGCGGATCTACTCACACGT
		GCCCACCGTGCCCAGCACCTCCGGTGGCCGGACCGTCAGT
		CTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCT
		CGCGAACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAA
		ACACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGAC
		GGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAG
		GAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCA
		CCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACA
		AGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCAATCGA
		GAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACC
		ACAGGTCTGCACCCTGCCCCCATCCCGGGAGGAGATGACC
		AAGAACCAGGTCAGCCTGAGCTGCGCGGTCAAAGGCTTCT
		ATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGC
		AGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGG
		ACTCCGACGGCTCCTTCTTCCTCGTTAGCAAGCTCACCGTG
		GACAAGAGCCGGTGGCAGCAGGGGAACGTCTTCTCATGCT
		CCGTGATGCATGAGGCTCTGCACAACCGCTTCACGCAGAA
		GAGCCTCTCCCTGTCTCCGGGTAAA
191	LM1953	CAAGTGCAGCTGCAGCAGTCAGGTCCAGGACTGGTGAAG
	STEAP2	CCCTCGCAGACCCTCTCACTCACCTGTGCCATCTCCGGGG
	Heavy Chain	ACAGTGTCTCCAGAAACAGTGCTGTTTGGAATTGGATCAG
	Knob (CD8)	GCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGAAGGAC
		ATACTACAGGTCCAAGTGGTATAATGATTATGCAGTATCT
		GTGAAAAGTCGAATAACCATCAACCCAGACACATCCAAG
		AACCAGTTCTCCCTGCAACTGAACTCTGTGACTCCCGAGG
		ACACGGCTGTGTATTACTGTGCAAGGGGGTTACGACAGAA
		CCAGTTCTACTACTACATGGACGTCTGGGGCAAAGGGACC
		ACGGTCACCGTCTCCTCCGCTAGCACCAAGGGACCCAGCG
		TGTTCCCTCTGGCACCTTCCAGCAAGTCTACCTCTGGCGGA
		ACAGCCGCTCTGGGCTGTCTGGTCAAGGACTACTTTCCCG
		AGCCTGTGACCGTGTCCTGGAATTCTGGCGCTCTGACCAG
		CGGAGTGCATACCTTTCCAGCTGTGCTGCAGTCCTCCGGC
		CTGTACTCTCTGTCCTCTGTCGTGACCGTGCCTTCCAGCTC
		TCTGGGCACCCAGACCTACATCTGCAATGTGAACCACAAG
		CCTTCCAACACCAAGGTGGACAAGAGGGTGGAACCCAAG
		TCCTGCGACAAGACCGGCGGATCTGAGGTCCAATTGTTGG
		AATCTGGAGGGGGGCTCGTGCAGCCGGGGGGAAGTTTGC
		GCCTTTCATGCGCTGCATCAGGGTTTACTTTCGACGACTAC
		GCTATTGGCTGGTTCAGGCAAGCGCCGGGCAAGGAACGC
		GAAGGCGTGTCCTGCATAAGAGTTTCAGATGGCAGTACCT
		ATTACGCCGATAGCGTCAAGGGACGATTCACGATCAGCCG
		GGATAATAGCAAAAATACACTTTATTTGCAAATGAACTCT
		CTTAGGGCAGAAGATACTGCGGTCTATTATTGTGCGGCCG
		GGTCCCTTTATACCTGCGTGCAGTCTATCGTATGGCCTGCT
		AGGCCCTATTACGATATGGATTACTGGGGACAGGGTACAC
		TCGTGACGGTATCCAGTGGAGGCGGCGGATCTACTCACAC
		GTGCCCACCGTGCCCAGCACCTCCGGTGGCCGGACCGTCA
		GTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGA
		TCTCCCGTACCCCTGAGGTCACATGCGTGGTGGTGGACGT
		GAAACACGAAGACCCTGAGGTCAAGTTCAACTGGTACGT
		GGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCG
		GGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGT
		CCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAG
		TACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCAA
		TCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAG
		AACCACAGGTCTACACCCTGCCCCCATGCCGGGAGGAGAT
		GACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGG
		CTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAAT
		GGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTG
		CTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCAC
		CGTGGACAAGAGCCGGTGGCAGCAGGGGAACGTCTTCTC
		ATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACG
		CAGAAGAGCTTAAGCCTGTCTCCGGGTAAA
192	LM1953	GAAATAGTGATGACGCAGTCTCCAGCCACCCTGTCTGTGT
	STEAP2	CTCCCGGGGAAAGAGCCACCCTCTCCTGCCGAGCCAGTCA
	Light Chain	GAGTGTTGCCAGCAACTTAGCCTGGTACCAGCAGAAACCT
		GGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCACCA
		GAGCCACTGGTATCCCAGCCAGGTTCAGTGGCAGTGGGTC
		TGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGTCT
		GAAGATTTTGCAGTTTATTACTGTCAACAGTATAATAACT
		GGCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAA
		ACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCAT
		CTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTG
		CCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAG
		TGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGG
		AGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACA
		GCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACG
		AGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGG
		GCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAG
		AGTGT
193	TPP-42197-	GAGGTGCAGCTGGTGGAAAGTGGAGGTGGCCTGGTTAAG
	BCMA	CCTGGCGGGAGCCTGCGCCTGAGCTGCGCTGCCAGCGGCT
	Heavy Chain	TCACTTTCTCATCTTACAGCATGAACTGGGTGCGCCAGGC
	Knob (CD8)	CCCTGGCAAAGGCCTTGAGTGGGTTAGTAGCATCAGCGGC
		CAGTCAAACTACATTTATTACGCCGATTCCGTCAAGGGCA
		GGTTTACAATCTCCCGTGATAATGCGAAGAATAGCCTCTA
		TCTCCAGATGAATTCCCTGCGCGCCGAAGATACAGCCGTG
		TATTACTGCGCGAGGGGCGGGAATTACTTTGTTGAATACT
		TCCAGTACTGGGGCCAGGGTACCCTCGTCACCGTTTCCAG
		TGCAAGCACCAAGGGCCCATCCGTCTTCCCCCTGGCACCC
		TCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCT
		GCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTC
		GTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTC
		CCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCA
		GCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGAC
		CTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAG
		GTGGACAAGAGAGTTGAACCAAAAAGCTGCGACAAGACC
		GGAGGATCTGAGGTCCAGCTGCTAGAGTCTGGCGGAGGA
		TTGGTTCAGCCTGGCGGATCTCTGAGACTGTCTTGTGCCGC
		CTCTGGCTTCACCTTCGACGATTACGCCATCGGCTGGTTCA
		GACAGGCCCCTGGCAAAGAGAGAGAGGGCGTCAGCTGCA
		TCAGAGTGTCTGACGGCTCCACCTACTACGCCGATTCTGT
		GAAGGGCAGATTCACCATCTCCCGCGACAACTCCAAGAAC
		ACCCTGTACCTGCAGATGAACTCCCTGCGGGCTGAGGACA
		CCGCCGTGTACTATTGTGCTGCCGGCTCTCTGTACACCTGT
		GTGCAGTCTATCGTGTGGCCCGCCAGACCTTACTACGACA
		TGGACTATTGGGGCCAGGGCACCCTAGTGACCGTCTCAAG
		CGGTGGCGGAGGATCTACCCACACCTGTCCTCCATGCCCT
		GCTCCACCCGTGGCCGGCCCATCCGTGTTTCTGTTCCCTCC
		AAAGCCTAAGGACACCCTGATGATCTCTCGGACCCCTGAA
		GTGACCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCG
		AAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGC
		ACAACGCCAAGACCAAGCCTAGAGAGGAACAGTACAACT
		CCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCA
		GGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTC
		CAACAAGGCCCTGCCTGCCCCCATCGAAAAGACCATCTCC
		AAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACC
		CTGCCACCTTGCCGGGAAGAGATGACCAAGAACCAGGTG
		TCCCTGTGGTGCCTCGTGAAGGGCTTCTACCCTTCCGATAT
		CGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAA
		CTACAAGACAACCCCTCCTGTGCTGGACTCCGACGGCTCA
		TTCTTCCTGTACTCCAAGCTGACAGTGGACAAGTCCAGAT
		GGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGA
		GGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTG
		TCCCCTGGAAAA
194	TPP-42197-	GAGGTGCAGCTGGTGGAAAGTGGAGGTGGCCTGGTTAAG
	BCMA	CCTGGCGGGAGCCTGCGCCTGAGCTGCGCTGCCAGCGGCT
	Heavy Chain	TCACTTTCTCATCTTACAGCATGAACTGGGTGCGCCAGGC
	Hole (TCR)	CCCTGGCAAAGGCCTTGAGTGGGTTAGTAGCATCAGCGGC
		CAGTCAAACTACATTTATTACGCCGATTCCGTCAAGGGCA
		GGTTTACAATCTCCCGTGATAATGCGAAGAATAGCCTCTA
		TCTCCAGATGAATTCCCTGCGCGCCGAAGATACAGCCGTG
		TATTACTGCGCGAGGGGCGGGAATTACTTTGTTGAATACT
		TCCAGTACTGGGGCCAGGGTACCCTCGTCACCGTTTCCAG
		TGCAAGCACCAAGGGCCCATCCGTCTTCCCCCTGGCACCC
		TCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCT
		GCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTC
		GTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTC
		CCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCA
		GCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGAC
		CTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAG
		GTGGACAAGAGAGTTGAACCAAAAAGCTGCGACAAGACC
		GGAGGATCTGAAGTGCAGCTGGTCGAGTCTGGCGGAGGA
		TTGGTTCAACCTGGCGGCTCCCTGAGACTGTCTTGTGTGGC
		TAGCGGAGATGTGCATAAGATCAATTTTCTCGGCTGGTAC
		AGACAGGCCCCTGGCAAAGAGAGAGAGAAGGTCGCCCAC
		ATCTCCATCGGCGACCAGACCGATTACGCCGACTCTGCCA
		AGGGCAGATTCACCATCTCTCGGGACGAGTCCAAGAACAT
		GGTGTACCTGCAGATGAACTCCCTGAAGCCTGAGGATACC
		GCCGTGTACTTCTGCCGGGCCTTCTCTCGGATCTACCCCTA
		CGATTATTGGGGCCAGGGCACCCTGGTCACAGTGTCATCT
		GGTGGCGGAGGATCTACCCACACCTGTCCTCCATGCCCTG
		CTCCACCCGTGGCCGGCCCTTCCGTGTTCCTGTTTCCTCCA
		AAGCCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAG
		TGACCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCGA
		AGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCAC
		AACGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCC
		ACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGG
		ATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCA
		ACAAGGCCCTGCCTGCCCCCATCGAAAAGACCATCTCTAA
		GGCTAAGGGCCAGCCTCGCGAGCCTCAAGTCTGTACACTG
		CCTCCTAGCCGGGAAGAGATGACCAAGAACCAGGTGTCC
		CTGTCTTGCGCCGTGAAGGGCTTCTACCCTTCCGATATCGC
		CGTGGAATGGGAGTCCAATGGCCAGCCTGAGAACAACTA
		CAAGACAACCCCTCCTGTGCTGGACTCCGACGGCTCATTC
		TTCCTGGTGTCCAAGCTGACAGTGGACAAGTCCAGATGGC
		AGCAGGGCAACGTGTTCTCCTGCTCTGTGATGCACGAGGC
		CCTGCACAACCGGTTCACCCAGAAATCCCTGTCTCTGTCC
		CCTGGCAAG
195	TPP-42197-	GAGATTGTCCTGACACAGTCCCCTGGCACCCTTAGCCTGT
	BCMA	CCCCCGGTGAAAGAGCGACCCTGTCCTGCCGTGCCTCACA
	Light Chain	ATACATCTCTTCCAACAATCTGGCGTGGTACCAACAGAAG
		CCCGGGCAAGCTCCCCGCCTCCTGATTTACGGGGCATCCA
		ATAGAGCCACCGGGATCCCTGATAGATTCTCCGGCAGCGG
		TAGCGGTACCGATTTCACCCTGACCATCTCTCGCCTGGAG
		CCTGAAGACTTCGCGGTGTACTATTGTCAACAGTATGCTG
		ATTCCCCAATCACATTCGGCCAGGGTACTAAGCTTGAAAT
		CAAGCGGACAGTGGCCGCACCGTCCGTGTTCATCTTCCCA
		CCTTCCGACGAGCAGCTGAAGTCTGGCACAGCCTCTGTCG
		TGTGCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGT
		GCAGTGGAAGGTGGACAATGCCCTGCAGTCCGGCAACTCC
		CAAGAGTCTGTGACCGAGCAGGACTCCAAGGACAGCACC
		TACAGCCTGTCCTCCACACTGACCCTGTCCAAGGCCGACT
		ACGAGAAGCACAAGGTGTACGCCTGCGAAGTGACCCATC
		AGGGCCTGTCTAGCCCTGTGACCAAGTCTTTCAACCGGGG
		CGAGTGC
196	LRC150016	GAAGTGCAGCTGGTTCAGTCTGGCGCCGAAGTGAAGAAA
	LRRC15	CCTGGCGCCTCTGTGAAGGTGTCCTGCAAGGCTTCCGGCT
	Heavy Chain	ACAAGTTCTCCAGCTACTGGATCGAGTGGGTCAAGCAGGC
	Knob (CD8)	TCCTGGACAGGGACTCGAGTGGATCGGAGAGATCCTGCCT
		GGCTCTGACACCACCAACTACAACGAGAAGTTCAAGGAC
		CGGGCCACCTTCACCTCCGACACCTCTATCAACACCGCCT
		ACATGGAACTGTCCCGGCTGAGATCTGACGACACCGCCGT
		GTACTACTGCGCCAGAGACAGAGGCAACTACAGAGCTTG
		GTTTGGCTACTGGGGCCAGGGCACACTGGTCACAGTTTCT
		TCTGCAAGCACCAAGGGACCCTCTGTGTTCCCTCTGGCTC
		CTTCCAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGG
		CTGCCTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTG
		TCCTGGAACTCTGGCGCTCTGACATCTGGCGTGCACACCT
		TTCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCC
		TCTGTCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGA
		CCTACATCTGCAATGTGAACCACAAGCCTTCCAACACCAA
		GGTGGACAAGAGAGTGGAACCCAAGTCCTGCGATAAGAC
		CGGCGGATCTGAGGTGCAGCTGTTGGAATCTGGCGGTGGA
		TTGGTTCAGCCTGGCGGCTCTCTGAGACTGTCTTGTGCCGC
		TTCTGGCTTCACCTTCGACGACTACGCCATCGGCTGGTTCA
		GACAGGCCCCTGGCAAAGAGAGAGAGGGCGTCAGCTGCA
		TCAGAGTGTCTGACGGCTCTACCTACTACGCCGACTCCGT
		GAAGGGCAGATTCACCATCTCTCGGGACAACTCCAAGAAC
		ACCCTGTACCTGCAGATGAACTCCCTGAGAGCCGAGGACA
		CCGCCGTGTACTATTGTGCTGCTGGCTCCCTGTACACCTGT
		GTGCAGTCTATCGTGTGGCCCGCCAGACCTTACTACGACA
		TGGATTATTGGGGCCAGGGCACCCTGGTCACAGTTTCTAG
		CGGAGGCGGAGGCAGCACCCATACCTGTCCTCCATGTCCT
		GCTCCTCCAGTGGCTGGCCCTTCCGTGTTTCTGTTCCCTCC
		AAAGCCTAAGGACACCCTGATGATCTCTCGGACCCCTGAA
		GTGACCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCG
		AAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGC
		ACAACGCCAAGACCAAGCCTAGAGAGGAACAGTACAACT
		CCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCA
		GGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTC
		CAACAAGGCCCTGCCTGCTCCTATCGAAAAGACCATCTCC
		AAGGCCAAGGGACAGCCCAGGGAACCCCAGGTTTACACC
		CTGCCTCCATGCCGGGAAGAGATGACCAAGAACCAGGTG
		TCCCTGTGGTGCCTGGTTAAGGGCTTCTACCCCTCCGATAT
		CGCCGTGGAATGGGAGTCTAATGGCCAGCCTGAGAACAA
		CTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCA
		TTCTTCCTGTACTCCAAGCTGACAGTGGACAAGTCCAGAT
		GGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGA
		GGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTG
		AGCCCCGGCAAA
197	LRC150016	GAAGTGCAGCTGGTTCAGTCTGGCGCCGAAGTGAAGAAA
	LRRC15	CCTGGCGCCTCTGTGAAGGTGTCCTGCAAGGCTTCCGGCT
	Heavy Chain	ACAAGTTCTCCAGCTACTGGATCGAGTGGGTCAAGCAGGC
	Hole (TCR)	TCCTGGACAGGGACTCGAGTGGATCGGAGAGATCCTGCCT
		GGCTCTGACACCACCAACTACAACGAGAAGTTCAAGGAC
		CGGGCCACCTTCACCTCCGACACCTCTATCAACACCGCCT
		ACATGGAACTGTCCCGGCTGAGATCTGACGACACCGCCGT
		GTACTACTGCGCCAGAGACAGAGGCAACTACAGAGCTTG
		GTTTGGCTACTGGGGCCAGGGCACACTGGTCACAGTTTCT
		TCTGCAAGTACTAAGGGCCCCTCTGTGTTCCCTCTGGCTCC
		TTCCAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGC
		TGCCTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGT
		CCTGGAACTCTGGCGCTCTGACATCTGGCGTGCACACCTT
		TCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCT
		CTGTCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGAC
		CTACATCTGCAATGTGAACCACAAGCCTTCCAACACCAAG
		GTGGACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACC
		GGCGGATCTGAGGTGCAGCTGGTGGAATCTGGCGGAGGA
		TTGGTTCAGCCTGGCGGCTCTCTGAGACTGTCTTGTGTGGC
		TAGCGGCGACGTGCACAAGATCAACTTTCTCGGCTGGTAC
		AGACAGGCCCCTGGCAAAGAGAGAGAGAAGGTCGCCCAC
		ATCTCCATCGGCGACCAGACCGATTACGCCGACTCTGCCA
		AGGGCAGATTCACCATCTCTCGGGACGAGTCCAAGAACAT
		GGTGTACCTGCAGATGAACTCCCTGAAGCCTGAGGATACC
		GCCGTGTACTTCTGCCGGGCCTTCTCTCGGATCTACCCCTA
		CGATTATTGGGGCCAGGGCACCCTGGTCACAGTTTCTAGC
		GGCGGAGGCGGCTCTACCCATACCTGTCCTCCATGTCCTG
		CTCCTCCAGTGGCTGGCCCATCCGTGTTTCTGTTCCCTCCA
		AAGCCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAG
		TGACCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCGA
		AGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCAC
		AACGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCC
		ACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGG
		ATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCA
		ACAAGGCCCTGCCTGCTCCTATCGAAAAGACCATCTCCAA
		GGCCAAGGGCCAGCCTCGGGAACCTCAAGTCTGTACCCTG
		CCTCCTAGCCGGGAAGAGATGACCAAGAACCAGGTGTCC
		CTGTCCTGTGCCGTGAAGGGCTTCTACCCTTCCGATATCGC
		CGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTA
		CAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCATTC
		TTCCTGGTGTCCAAGCTGACAGTGGACAAGTCCAGATGGC
		AGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGC
		CCTGCACAACAGGTTCACCCAGAAGTCCCTGTCTCTGAGC
		CCTGGCAAG
198	LRC150016	GACATCCAGATGACCCAGTCTCCATCCTCTCTGTCCGCCTC
	LRRC15	TGTGGGCGACAGAGTGACCATCACCTGTAGAGCCAGCCA
	Light Chain	GGACATCTCCAACTACCTGAACTGGTATCAGCAGAAACCT
		GGCGGCGCTGTGAAGTTCCTGATCTACTACACCTCTCGGC
		TGCACTCCGGCGTGCCCTCTAGATTTTCTGGCTCTGGATCC
		GGCACCGACTATACCCTGACAATCTCCAGCCTGCAGCCTG
		AGGACTTCGCTACCTACTTCTGCCAGCAAGGCGAGGCTCT
		GCCTTGGACATTTGGCGGCGGAACAAAGGTGGAAATCAA
		ACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCAT
		CTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTG
		CCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAG
		TGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGG
		AGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACA
		GCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACG
		AGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGG
		GCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAG
		AGTGT
199	TPP-46956	GAGGTGCAGTTGGTCCAATCAGGAGCAGAAGTCAAAAAA
	LRRC15	CCCGGTGCTTCCGTTAAAGTATCTTGCAAAGCTAGCGGGT
	Heavy Chain	ACAAGTTCTCTTCCTATTGGATCGAGTGGGTTAAGCAGGC
	Knob (CD8)	ACCTGGACAGGGTCTTGAATGGATAGGCGAAATCCTTCCC
		GGTAGCGACACCACAAACTATGCTCAAAAGTTCCAAGATA
		GAGTTACTTTCACTTCAGATACTTCCATATCTACTGCATAT
		ATGGAACTGTCCCGTTTGAGGAGTGATGACACAGCAGTGT
		ACTACTGTGCCCGTGACCGTGGAAACTACCGTGCATGGTT
		CGGCTATTGGGGTCAGGGTACACTCGTAACAGTGAGCTCA
		GCAAGCACCAAGGGACCCTCTGTGTTCCCTCTGGCTCCTT
		CCAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGCTG
		CCTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGTCCT
		GGAACTCTGGCGCTCTGACATCTGGCGTGCACACCTTTCC
		AGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCTCTG
		TCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGACCTA
		CATCTGCAATGTGAACCACAAGCCTTCCAACACCAAGGTG
		GACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACCGGC
		GGATCTGAGGTACAGCTGCTGGAATCAGGAGGCGGCTTG
		GTTCAGCCAGGAGGCTCACTTCGGCTCTCCTGCGCAGCAA
		GTGGGTTTACATTTACTGATTATGCTATAGGCTGGTTTCGG
		CAAGCACCTGGTAAGGAACGCGAAGGTGTGTCATGTATTC
		GCGTAAGCGACGGATCAACATACTATGCAGACAGTGTCA
		AAGGTAGATTTACTATTAGTCGTGACAGCTCAAAGAATAC
		TCTGTACTTGCAAATGAATTCCCTTCGTGCCGAAGATACT
		GCCGTTTACTATTGCGCCGCAGGATCACTCTACACCTGCG
		TGCAGAGCATTGTTTGGCCTGCCAGGCCTTATTATGATAT
		GGACTATTGGGGCCAAGGCACTTTGGTCACTGTTAGCTCT
		GGAGGCGGAGGCAGCACCCATACCTGTCCTCCATGTCCTG
		CTCCTCCAGTGGCTGGCCCTTCCGTGTTTCTGTTCCCTCCA
		AAGCCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAG
		TGACCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCGA
		AGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCAC
		AACGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCC
		ACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGG
		ATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCA
		ACAAGGCCCTGCCTGCTCCTATCGAAAAGACCATCTCCAA
		GGCCAAGGGACAGCCCAGGGAACCCCAGGTTTACACCCT
		GCCTCCATGCCGGGAAGAGATGACCAAGAACCAGGTGTC
		CCTGTGGTGCCTGGTTAAGGGCTTCTACCCCTCCGATATCG
		CCGTGGAATGGGAGTCTAATGGCCAGCCTGAGAACAACT
		ACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCATT
		CTTCCTGTACTCCAAGCTGACAGTGGACAAGTCCAGATGG
		CAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGG
		CCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGAG
		CCCCGGCAAA
200	TPP-46956	GAGGTGCAGTTGGTCCAATCAGGAGCAGAAGTCAAAAAA
	LRRC15	CCCGGTGCTTCCGTTAAAGTATCTTGCAAAGCTAGCGGGT
	Heavy Chain	ACAAGTTCTCTTCCTATTGGATCGAGTGGGTTAAGCAGGC
	Hole (TCR)	ACCTGGACAGGGTCTTGAATGGATAGGCGAAATCCTTCCC
		GGTAGCGACACCACAAACTATGCTCAAAAGTTCCAAGATA
		GAGTTACTTTCACTTCAGATACTTCCATATCTACTGCATAT
		ATGGAACTGTCCCGTTTGAGGAGTGATGACACAGCAGTGT
		ACTACTGTGCCCGTGACCGTGGAAACTACCGTGCATGGTT
		CGGCTATTGGGGTCAGGGTACACTCGTAACAGTGAGCTCA
		GCAAGTACTAAGGGCCCCTCTGTGTTCCCTCTGGCTCCTTC
		CAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGCTGC
		CTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGTCCT
		GGAACTCTGGCGCTCTGACATCTGGCGTGCACACCTTTCC
		AGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCTCTG
		TCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGACCTA
		CATCTGCAATGTGAACCACAAGCCTTCCAACACCAAGGTG
		GACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACCGGC
		GGATCTGAGGTGCAGCTGGTGGAATCTGGCGGAGGATTG
		GTTCAGCCTGGCGGCTCTCTGAGACTGTCTTGTGTGGCTA
		GCGGCGACGTGCACAAGATCAACTTTCTCGGCTGGTACAG
		ACAGGCCCCTGGCAAAGAGAGAGAGAAGGTCGCCCACAT
		CTCCATCGGCGACCAGACCGATTACGCCGACTCTGCCAAG
		GGCAGATTCACCATCTCTCGGGACGAGTCCAAGAACATGG
		TGTACCTGCAGATGAACTCCCTGAAGCCTGAGGATACCGC
		CGTGTACTTCTGCCGGGCCTTCTCTCGGATCTACCCCTACG
		ATTATTGGGGCCAGGGCACCCTGGTCACAGTTTCTAGCGG
		CGGAGGCGGCTCTACCCATACCTGTCCTCCATGTCCTGCTC
		CTCCAGTGGCTGGCCCATCCGTGTTTCTGTTCCCTCCAAAG
		CCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAGTGA
		CCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCGAAGT
		GAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAA
		CGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCAC
		CTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGAT
		TGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAAC
		AAGGCCCTGCCTGCTCCTATCGAAAAGACCATCTCCAAGG
		CCAAGGGCCAGCCTCGGGAACCTCAAGTCTGTACCCTGCC
		TCCTAGCCGGGAAGAGATGACCAAGAACCAGGTGTCCCT
		GTCCTGTGCCGTGAAGGGCTTCTACCCTTCCGATATCGCC
		GTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTAC
		AAGACCACACCTCCTGTGCTGGACTCCGACGGCTCATTCT
		TCCTGGTGTCCAAGCTGACAGTGGACAAGTCCAGATGGCA
		GCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCC
		CTGCACAACAGGTTCACCCAGAAGTCCCTGTCTCTGAGCC
		CTGGCAAG
201	TPP-46956	GACATCCAGATGACCCAGAGTCCCTCTAGTCTCTCAGCTT
	LRRC15	CCGTGGGGGACCGCGTCACTATAACTTGCCGTGCCTCACA
	Light Chain	AGACATATCAAACTACCTCAACTGGTATCAACAAAAGCCT
		GGAAAAGCTCCTAAGTTTTTGATTTATTACACTTCCAGGCT
		GGAGTCCGGTGTCCCATCCAGGTTCAGCGGCAGTGGAAGC
		GGAACCGACTATACACTGACTATTTCTAGCTTGCAACCCG
		AGGACTTTGCCACCTATTATTGCCAGCAAGGTAACGCCCT
		GCCCTGGACCTTCGGTGGCGGAACCAAGGTGGAAATCAA
		GCGGACAGTGGCCGCACCGTCCGTGTTCATCTTCCCACCT
		TCCGACGAGCAGCTGAAGTCTGGCACAGCCTCTGTCGTGT
		GCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCA
		GTGGAAGGTGGACAATGCCCTGCAGTCCGGCAACTCCCAA
		GAGTCTGTGACCGAGCAGGACTCCAAGGACAGCACCTAC
		AGCCTGTCCTCCACACTGACCCTGTCCAAGGCCGACTACG
		AGAAGCACAAGGTGTACGCCTGCGAAGTGACCCATCAGG
		GCCTGTCTAGCCCTGTGACCAAGTCTTTCAACCGGGGCGA
		GTGC
202	TPP-46957	GAGGTTCAGTTGGTACAATCTGGAGCCGAGGTGAAAAAA
	LRRC15	CCAGGTGCAAGCGTCAAGGTTTCCTGCAAAGCTAGTGGGT
	Heavy Chain	ACAAATTCAGTTCATACTGGATAGAGTGGGTTAAGCAAGC
	Knob (CD8)	TCCTGGACAAGGTTTGGAGTGGATTGGTGAAATCTTGCCT
		GGCAGCGATACTACAAACTACGCTCAGCAGTTTCAGGACC
		GCGTAACATTCACCTCTGATACATCTATATCAACTGCATAT
		ATGGAACTTAGCAGGCTCAGGTCCGACGACACTGCCGTCT
		ATTACTGCGCTCGGGATAGGGGGAATTATCGTGCTTGGTT
		TGGTTATTGGGGTCAAGGGACACTGGTTACAGTTTCTTCA
		GCAAGCACCAAGGGACCCTCTGTGTTCCCTCTGGCTCCTT
		CCAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGCTG
		CCTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGTCCT
		GGAACTCTGGCGCTCTGACATCTGGCGTGCACACCTTTCC
		AGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCTCTG
		TCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGACCTA
		CATCTGCAATGTGAACCACAAGCCTTCCAACACCAAGGTG
		GACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACCGGC
		GGATCTGAGGTACAGCTGCTGGAATCAGGAGGCGGCTTG
		GTTCAGCCAGGAGGCTCACTTCGGCTCTCCTGCGCAGCAA
		GTGGGTTTACATTTACTGATTATGCTATAGGCTGGTTTCGG
		CAAGCACCTGGTAAGGAACGCGAAGGTGTGTCATGTATTC
		GCGTAAGCGACGGATCAACATACTATGCAGACAGTGTCA
		AAGGTAGATTTACTATTAGTCGTGACAGCTCAAAGAATAC
		TCTGTACTTGCAAATGAATTCCCTTCGTGCCGAAGATACT
		GCCGTTTACTATTGCGCCGCAGGATCACTCTACACCTGCG
		TGCAGAGCATTGTTTGGCCTGCCAGGCCTTATTATGATAT
		GGACTATTGGGGCCAAGGCACTTTGGTCACTGTTAGCTCT
		GGAGGCGGAGGCAGCACCCATACCTGTCCTCCATGTCCTG
		CTCCTCCAGTGGCTGGCCCTTCCGTGTTTCTGTTCCCTCCA
		AAGCCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAG
		TGACCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCGA
		AGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCAC
		AACGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCC
		ACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGG
		ATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCA
		ACAAGGCCCTGCCTGCTCCTATCGAAAAGACCATCTCCAA
		GGCCAAGGGACAGCCCAGGGAACCCCAGGTTTACACCCT
		GCCTCCATGCCGGGAAGAGATGACCAAGAACCAGGTGTC
		CCTGTGGTGCCTGGTTAAGGGCTTCTACCCCTCCGATATCG
		CCGTGGAATGGGAGTCTAATGGCCAGCCTGAGAACAACT
		ACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCATT
		CTTCCTGTACTCCAAGCTGACAGTGGACAAGTCCAGATGG
		CAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGG
		CCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGAG
		CCCCGGCAAA
203	TPP-46957	GAGGTTCAGTTGGTACAATCTGGAGCCGAGGTGAAAAAA
	LRRC15	CCAGGTGCAAGCGTCAAGGTTTCCTGCAAAGCTAGTGGGT
	Heavy Chain	ACAAATTCAGTTCATACTGGATAGAGTGGGTTAAGCAAGC
	Hole (TCR)	TCCTGGACAAGGTTTGGAGTGGATTGGTGAAATCTTGCCT
		GGCAGCGATACTACAAACTACGCTCAGCAGTTTCAGGACC
		GCGTAACATTCACCTCTGATACATCTATATCAACTGCATAT
		ATGGAACTTAGCAGGCTCAGGTCCGACGACACTGCCGTCT
		ATTACTGCGCTCGGGATAGGGGGAATTATCGTGCTTGGTT
		TGGTTATTGGGGTCAAGGGACACTGGTTACAGTTTCTTCA
		GCAAGTACTAAGGGCCCCTCTGTGTTCCCTCTGGCTCCTTC
		CAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGCTGC
		CTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGTCCT
		GGAACTCTGGCGCTCTGACATCTGGCGTGCACACCTTTCC
		AGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCTCTG
		TCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGACCTA
		CATCTGCAATGTGAACCACAAGCCTTCCAACACCAAGGTG
		GACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACCGGC
		GGATCTGAGGTGCAGCTGGTGGAATCTGGCGGAGGATTG
		GTTCAGCCTGGCGGCTCTCTGAGACTGTCTTGTGTGGCTA
		GCGGCGACGTGCACAAGATCAACTTTCTCGGCTGGTACAG
		ACAGGCCCCTGGCAAAGAGAGAGAGAAGGTCGCCCACAT
		CTCCATCGGCGACCAGACCGATTACGCCGACTCTGCCAAG
		GGCAGATTCACCATCTCTCGGGACGAGTCCAAGAACATGG
		TGTACCTGCAGATGAACTCCCTGAAGCCTGAGGATACCGC
		CGTGTACTTCTGCCGGGCCTTCTCTCGGATCTACCCCTACG
		ATTATTGGGGCCAGGGCACCCTGGTCACAGTTTCTAGCGG
		CGGAGGCGGCTCTACCCATACCTGTCCTCCATGTCCTGCTC
		CTCCAGTGGCTGGCCCATCCGTGTTTCTGTTCCCTCCAAAG
		CCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAGTGA
		CCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCGAAGT
		GAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAA
		CGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCAC
		CTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGAT
		TGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAAC
		AAGGCCCTGCCTGCTCCTATCGAAAAGACCATCTCCAAGG
		CCAAGGGCCAGCCTCGGGAACCTCAAGTCTGTACCCTGCC
		TCCTAGCCGGGAAGAGATGACCAAGAACCAGGTGTCCCT
		GTCCTGTGCCGTGAAGGGCTTCTACCCTTCCGATATCGCC
		GTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTAC
		AAGACCACACCTCCTGTGCTGGACTCCGACGGCTCATTCT
		TCCTGGTGTCCAAGCTGACAGTGGACAAGTCCAGATGGCA
		GCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCC
		CTGCACAACAGGTTCACCCAGAAGTCCCTGTCTCTGAGCC
		CTGGCAAG
204	TPP-46957	GATATTCAGATGACACAATCCCCTAGCAGTCTCAGTGCCT
	LRRC15	CAGTAGGGGACCGCGTCACAATAACATGTAGAGCCTCCCA
	Light Chain	GGATATTTCTAACTACTTGAACTGGTACCAGCAAAAGCCC
		GGTAAAGCCCCTAAATTCCTTATTTACTACACAAGCCGAT
		TGCACTCTGGTGTACCATCCAGGTTTAGTGGAAGCGGCTC
		TGGTACAGATTACACACTGACCATTTCATCCCTGCAACCA
		GAGGATTTCGCCACATATTACTGCCAACAGGGAAACGCCC
		TCCCTTGGACCTTTGGTGGGGGAACTAAAGTTGAGATCAA
		GCGGACAGTGGCCGCACCGTCCGTGTTCATCTTCCCACCT
		TCCGACGAGCAGCTGAAGTCTGGCACAGCCTCTGTCGTGT
		GCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCA
		GTGGAAGGTGGACAATGCCCTGCAGTCCGGCAACTCCCAA
		GAGTCTGTGACCGAGCAGGACTCCAAGGACAGCACCTAC
		AGCCTGTCCTCCACACTGACCCTGTCCAAGGCCGACTACG
		AGAAGCACAAGGTGTACGCCTGCGAAGTGACCCATCAGG
		GCCTGTCTAGCCCTGTGACCAAGTCTTTCAACCGGGGCGA
		GTGC
205	TPP-46958	GAGGTGCAGCTTGTTCAAAGCGGTGCAGAGGTTAAGAAG
	LRRC15	CCTGGAGCTAGCGTAAAAGTGTCCTGCAAAGCTAGTGGGT
	Heavy Chain	ATAAGTTCTCTAGCTACTGGATCGAGTGGGTGAAACAAGC
	Knob (CD8)	CCCAGGGCAGGGGTTGGAGTGGATTGGAGAAATACTCCC
		CGGATCAGACGCCACTAACTATGCACAGAAGTTTCAAGAC
		AGGGTTACCTTCACATCAGACACATCCATATCAACCGCAT
		ATATGGAGTTGTCCAGGCTGCGGAGCGACGACACCGCTGT
		CTATTATTGTGCCAGGGACCGGGGGAACTACAGGGCTTGG
		TTTGGGTACTGGGGACAGGGTACACTCGTCACAGTGTCCT
		CCGCAAGCACCAAGGGACCCTCTGTGTTCCCTCTGGCTCC
		TTCCAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGC
		TGCCTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGT
		CCTGGAACTCTGGCGCTCTGACATCTGGCGTGCACACCTT
		TCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCT
		CTGTCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGAC
		CTACATCTGCAATGTGAACCACAAGCCTTCCAACACCAAG
		GTGGACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACC
		GGCGGATCTGAGGTACAGCTGCTGGAATCAGGAGGCGGC
		TTGGTTCAGCCAGGAGGCTCACTTCGGCTCTCCTGCGCAG
		CAAGTGGGTTTACATTTACTGATTATGCTATAGGCTGGTTT
		CGGCAAGCACCTGGTAAGGAACGCGAAGGTGTGTCATGT
		ATTCGCGTAAGCGACGGATCAACATACTATGCAGACAGTG
		TCAAAGGTAGATTTACTATTAGTCGTGACAGCTCAAAGAA
		TACTCTGTACTTGCAAATGAATTCCCTTCGTGCCGAAGAT
		ACTGCCGTTTACTATTGCGCCGCAGGATCACTCTACACCT
		GCGTGCAGAGCATTGTTTGGCCTGCCAGGCCTTATTATGA
		TATGGACTATTGGGGCCAAGGCACTTTGGTCACTGTTAGC
		TCTGGAGGCGGAGGCAGCACCCATACCTGTCCTCCATGTC
		CTGCTCCTCCAGTGGCTGGCCCTTCCGTGTTTCTGTTCCCT
		CCAAAGCCTAAGGACACCCTGATGATCTCTCGGACCCCTG
		AAGTGACCTGCGTGGTGGTGGATGTGAAGCACGAGGATC
		CCGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGT
		GCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTACAA
		CTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCAC
		CAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTG
		TCCAACAAGGCCCTGCCTGCTCCTATCGAAAAGACCATCT
		CCAAGGCCAAGGGACAGCCCAGGGAACCCCAGGTTTACA
		CCCTGCCTCCATGCCGGGAAGAGATGACCAAGAACCAGG
		TGTCCCTGTGGTGCCTGGTTAAGGGCTTCTACCCCTCCGAT
		ATCGCCGTGGAATGGGAGTCTAATGGCCAGCCTGAGAAC
		AACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCT
		CATTCTTCCTGTACTCCAAGCTGACAGTGGACAAGTCCAG
		ATGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCAC
		GAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTC
		TGAGCCCCGGCAAA
206	TPP-46958	GAGGTGCAGCTTGTTCAAAGCGGTGCAGAGGTTAAGAAG
	LRRC15	CCTGGAGCTAGCGTAAAAGTGTCCTGCAAAGCTAGTGGGT
	Heavy Chain	ATAAGTTCTCTAGCTACTGGATCGAGTGGGTGAAACAAGC
	Hole (TCR)	CCCAGGGCAGGGGTTGGAGTGGATTGGAGAAATACTCCC
		CGGATCAGACGCCACTAACTATGCACAGAAGTTTCAAGAC
		AGGGTTACCTTCACATCAGACACATCCATATCAACCGCAT
		ATATGGAGTTGTCCAGGCTGCGGAGCGACGACACCGCTGT
		CTATTATTGTGCCAGGGACCGGGGGAACTACAGGGCTTGG
		TTTGGGTACTGGGGACAGGGTACACTCGTCACAGTGTCCT
		CCGCAAGTACTAAGGGCCCCTCTGTGTTCCCTCTGGCTCCT
		TCCAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGCT
		GCCTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGTC
		CTGGAACTCTGGCGCTCTGACATCTGGCGTGCACACCTTT
		CCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCTC
		TGTCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGACC
		TACATCTGCAATGTGAACCACAAGCCTTCCAACACCAAGG
		TGGACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACCG
		GCGGATCTGAGGTGCAGCTGGTGGAATCTGGCGGAGGATT
		GGTTCAGCCTGGCGGCTCTCTGAGACTGTCTTGTGTGGCT
		AGCGGCGACGTGCACAAGATCAACTTTCTCGGCTGGTACA
		GACAGGCCCCTGGCAAAGAGAGAGAGAAGGTCGCCCACA
		TCTCCATCGGCGACCAGACCGATTACGCCGACTCTGCCAA
		GGGCAGATTCACCATCTCTCGGGACGAGTCCAAGAACATG
		GTGTACCTGCAGATGAACTCCCTGAAGCCTGAGGATACCG
		CCGTGTACTTCTGCCGGGCCTTCTCTCGGATCTACCCCTAC
		GATTATTGGGGCCAGGGCACCCTGGTCACAGTTTCTAGCG
		GCGGAGGCGGCTCTACCCATACCTGTCCTCCATGTCCTGC
		TCCTCCAGTGGCTGGCCCATCCGTGTTTCTGTTCCCTCCAA
		AGCCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAGT
		GACCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCGA
		AGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCAC
		AACGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCC
		ACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGG
		ATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCA
		ACAAGGCCCTGCCTGCTCCTATCGAAAAGACCATCTCCAA
		GGCCAAGGGCCAGCCTCGGGAACCTCAAGTCTGTACCCTG
		CCTCCTAGCCGGGAAGAGATGACCAAGAACCAGGTGTCC
		CTGTCCTGTGCCGTGAAGGGCTTCTACCCTTCCGATATCGC
		CGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTA
		CAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCATTC
		TTCCTGGTGTCCAAGCTGACAGTGGACAAGTCCAGATGGC
		AGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGC
		CCTGCACAACAGGTTCACCCAGAAGTCCCTGTCTCTGAGC
		CCTGGCAAG
207	TPP-46958	GATATTCAGATGACACAATCCCCTAGCAGTCTCAGTGCCT
	LRRC15	CAGTAGGGGACCGCGTCACAATAACATGTAGAGCCTCCCA
	Light Chain	GGATATTTCTAACTACTTGAACTGGTACCAGCAAAAGCCC
		GGTAAAGCCCCTAAATTCCTTATTTACTACACAAGCCGAT
		TGCACTCTGGTGTACCATCCAGGTTTAGTGGAAGCGGCTC
		TGGTACAGATTACACACTGACCATTTCATCCCTGCAACCA
		GAGGATTTCGCCACATATTACTGCCAACAGGGAAACGCCC
		TCCCTTGGACCTTTGGTGGGGGAACTAAAGTTGAGATCAA
		GCGGACAGTGGCCGCACCGTCCGTGTTCATCTTCCCACCT
		TCCGACGAGCAGCTGAAGTCTGGCACAGCCTCTGTCGTGT
		GCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCA
		GTGGAAGGTGGACAATGCCCTGCAGTCCGGCAACTCCCAA
		GAGTCTGTGACCGAGCAGGACTCCAAGGACAGCACCTAC
		AGCCTGTCCTCCACACTGACCCTGTCCAAGGCCGACTACG
		AGAAGCACAAGGTGTACGCCTGCGAAGTGACCCATCAGG
		GCCTGTCTAGCCCTGTGACCAAGTCTTTCAACCGGGGCGA
		GTGC
208	TPP-46959	GAGGTGCAGTTGGTCCAATCAGGAGCAGAAGTCAAAAAA
	LRRC15	CCCGGTGCTTCCGTTAAAGTATCTTGCAAAGCTAGCGGGT
	Heavy Chain	ACAAGTTCTCTTCCTATTGGATCGAGTGGGTTAAGCAGGC
	Knob (CD8)	ACCTGGACAGGGTCTTGAATGGATAGGCGAAATCCTTCCC
		GGTAGCGACACCACAAACTATGCTCAAAAGTTCCAAGATA
		GAGTTACTTTCACTTCAGATACTTCCATATCTACTGCATAT
		ATGGAACTGTCCCGTTTGAGGAGTGATGACACAGCAGTGT
		ACTACTGTGCCCGTGACCGTGGAAACTACCGTGCATGGTT
		CGGCTATTGGGGTCAGGGTACACTCGTAACAGTGAGCTCA
		GCAAGCACCAAGGGACCCTCTGTGTTCCCTCTGGCTCCTT
		CCAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGCTG
		CCTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGTCCT
		GGAACTCTGGCGCTCTGACATCTGGCGTGCACACCTTTCC
		AGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCTCTG
		TCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGACCTA
		CATCTGCAATGTGAACCACAAGCCTTCCAACACCAAGGTG
		GACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACCGGC
		GGATCTGAGGTACAGCTGCTGGAATCAGGAGGCGGCTTG
		GTTCAGCCAGGAGGCTCACTTCGGCTCTCCTGCGCAGCAA
		GTGGGTTTACATTTACTGATTATGCTATAGGCTGGTTTCGG
		CAAGCACCTGGTAAGGAACGCGAAGGTGTGTCATGTATTC
		GCGTAAGCGACGGATCAACATACTATGCAGACAGTGTCA
		AAGGTAGATTTACTATTAGTCGTGACAGCTCAAAGAATAC
		TCTGTACTTGCAAATGAATTCCCTTCGTGCCGAAGATACT
		GCCGTTTACTATTGCGCCGCAGGATCACTCTACACCTGCG
		TGCAGAGCATTGTTTGGCCTGCCAGGCCTTATTATGATAT
		GGACTATTGGGGCCAAGGCACTTTGGTCACTGTTAGCTCT
		GGAGGCGGAGGCAGCACCCATACCTGTCCTCCATGTCCTG
		CTCCTCCAGTGGCTGGCCCTTCCGTGTTTCTGTTCCCTCCA
		AAGCCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAG
		TGACCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCGA
		AGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCAC
		AACGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCC
		ACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGG
		ATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCA
		ACAAGGCCCTGCCTGCTCCTATCGAAAAGACCATCTCCAA
		GGCCAAGGGACAGCCCAGGGAACCCCAGGTTTACACCCT
		GCCTCCATGCCGGGAAGAGATGACCAAGAACCAGGTGTC
		CCTGTGGTGCCTGGTTAAGGGCTTCTACCCCTCCGATATCG
		CCGTGGAATGGGAGTCTAATGGCCAGCCTGAGAACAACT
		ACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCATT
		CTTCCTGTACTCCAAGCTGACAGTGGACAAGTCCAGATGG
		CAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGG
		CCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGAG
		CCCCGGCAAA
209	TPP-46959	AGGTGCAGTTGGTCCAATCAGGAGCAGAAGTCAAAAAAC
	LRRC15	CCGGTGCTTCCGTTAAAGTATCTTGCAAAGCTAGCGGGTA
	Heavy Chain	CAAGTTCTCTTCCTATTGGATCGAGTGGGTTAAGCAGGCA
	Hole (TCR)	CCTGGACAGGGTCTTGAATGGATAGGCGAAATCCTTCCCG
		GTAGCGACACCACAAACTATGCTCAAAAGTTCCAAGATAG
		AGTTACTTTCACTTCAGATACTTCCATATCTACTGCATATA
		TGGAACTGTCCCGTTTGAGGAGTGATGACACAGCAGTGTA
		CTACTGTGCCCGTGACCGTGGAAACTACCGTGCATGGTTC
		GGCTATTGGGGTCAGGGTACACTCGTAACAGTGAGCTCAG
		CAAGTACTAAGGGCCCCTCTGTGTTCCCTCTGGCTCCTTCC
		AGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGCTGCC
		TGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGTCCTG
		GAACTCTGGCGCTCTGACATCTGGCGTGCACACCTTTCCA
		GCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCTCTGT
		CGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGACCTAC
		ATCTGCAATGTGAACCACAAGCCTTCCAACACCAAGGTGG
		ACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACCGGCG
		GATCTGAGGTGCAGCTGGTGGAATCTGGCGGAGGATTGGT
		TCAGCCTGGCGGCTCTCTGAGACTGTCTTGTGTGGCTAGC
		GGCGACGTGCACAAGATCAACTTTCTCGGCTGGTACAGAC
		AGGCCCCTGGCAAAGAGAGAGAGAAGGTCGCCCACATCT
		CCATCGGCGACCAGACCGATTACGCCGACTCTGCCAAGGG
		CAGATTCACCATCTCTCGGGACGAGTCCAAGAACATGGTG
		TACCTGCAGATGAACTCCCTGAAGCCTGAGGATACCGCCG
		TGTACTTCTGCCGGGCCTTCTCTCGGATCTACCCCTACGAT
		TATTGGGGCCAGGGCACCCTGGTCACAGTTTCTAGCGGCG
		GAGGCGGCTCTACCCATACCTGTCCTCCATGTCCTGCTCCT
		CCAGTGGCTGGCCCATCCGTGTTTCTGTTCCCTCCAAAGCC
		TAAGGACACCCTGATGATCTCTCGGACCCCTGAAGTGACC
		TGCGTGGTGGTGGATGTGAAGCACGAGGATCCCGAAGTG
		AAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAAC
		GCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCACC
		TACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATT
		GGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACA
		AGGCCCTGCCTGCTCCTATCGAAAAGACCATCTCCAAGGC
		CAAGGGCCAGCCTCGGGAACCTCAAGTCTGTACCCTGCCT
		CCTAGCCGGGAAGAGATGACCAAGAACCAGGTGTCCCTG
		TCCTGTGCCGTGAAGGGCTTCTACCCTTCCGATATCGCCGT
		GGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAA
		GACCACACCTCCTGTGCTGGACTCCGACGGCTCATTCTTCC
		TGGTGTCCAAGCTGACAGTGGACAAGTCCAGATGGCAGC
		AGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCT
		GCACAACAGGTTCACCCAGAAGTCCCTGTCTCTGAGCCCT
		GGCAAG
210	TPP-46959	GACATCCAGATGACCCAGAGTCCCTCTAGTCTCTCAGCTT
	LRRC15	CCGTGGGGGACCGCGTCACTATAACTTGCCGTGCCTCACA
	Light Chain	AGACATATCAAACTACCTCAACTGGTATCAACAAAAGCCT
		GGAAAAGCTCCTAAGTTTTTGATTTATTACACTTCCAGGCT
		GGAGTCCGGTGTCCCATCCAGGTTCAGCGGCAGTGGAAGC
		GGAACCGACTATACACTGACTATTTCTAGCTTGCAACCCG
		AGGACTTTGCCACCTATTATTGCCAGCAAGGTAACGCCCT
		GCCCTGGACCTTCGGTGGCGGAACCAAGGTGGAAATCAA
		GCGGACAGTGGCCGCACCGTCCGTGTTCATCTTCCCACCT
		TCCGACGAGCAGCTGAAGTCTGGCACAGCCTCTGTCGTGT
		GCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCA
		GTGGAAGGTGGACAATGCCCTGCAGTCCGGCAACTCCCAA
		GAGTCTGTGACCGAGCAGGACTCCAAGGACAGCACCTAC
		AGCCTGTCCTCCACACTGACCCTGTCCAAGGCCGACTACG
		AGAAGCACAAGGTGTACGCCTGCGAAGTGACCCATCAGG
		GCCTGTCTAGCCCTGTGACCAAGTCTTTCAACCGGGGCGA
		GTGC
211	TPP-46960	GAGGTGCAGTTGGTCCAATCAGGAGCAGAAGTCAAAAAA
	LRRC15	CCCGGTGCTTCCGTTAAAGTATCTTGCAAAGCTAGCGGGT
	Heavy Chain	ACAAGTTCTCTTCCTATTGGATCGAGTGGGTTAAGCAGGC
	Knob (CD8)	ACCTGGACAGGGTCTTGAATGGATAGGCGAAATCCTTCCC
		GGTAGCGACACCACAAACTATGCTCAAAAGTTCCAAGATA
		GAGTTACTTTCACTTCAGATACTTCCATATCTACTGCATAT
		ATGGAACTGTCCCGTTTGAGGAGTGATGACACAGCAGTGT
		ACTACTGTGCCCGTGACCGTGGAAACTACCGTGCATGGTT
		CGGCTATTGGGGTCAGGGTACACTCGTAACAGTGAGCTCA
		GCAAGCACCAAGGGACCCTCTGTGTTCCCTCTGGCTCCTT
		CCAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGCTG
		CCTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGTCCT
		GGAACTCTGGCGCTCTGACATCTGGCGTGCACACCTTTCC
		AGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCTCTG
		TCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGACCTA
		CATCTGCAATGTGAACCACAAGCCTTCCAACACCAAGGTG
		GACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACCGGC
		GGATCTGAGGTACAGCTGCTGGAATCAGGAGGCGGCTTG
		GTTCAGCCAGGAGGCTCACTTCGGCTCTCCTGCGCAGCAA
		GTGGGTTTACATTTACTGATTATGCTATAGGCTGGTTTCGG
		CAAGCACCTGGTAAGGAACGCGAAGGTGTGTCATGTATTC
		GCGTAAGCGACGGATCAACATACTATGCAGACAGTGTCA
		AAGGTAGATTTACTATTAGTCGTGACAGCTCAAAGAATAC
		TCTGTACTTGCAAATGAATTCCCTTCGTGCCGAAGATACT
		GCCGTTTACTATTGCGCCGCAGGATCACTCTACACCTGCG
		TGCAGAGCATTGTTTGGCCTGCCAGGCCTTATTATGATAT
		GGACTATTGGGGCCAAGGCACTTTGGTCACTGTTAGCTCT
		GGAGGCGGAGGCAGCACCCATACCTGTCCTCCATGTCCTG
		CTCCTCCAGTGGCTGGCCCTTCCGTGTTTCTGTTCCCTCCA
		AAGCCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAG
		TGACCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCGA
		AGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCAC
		AACGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCC
		ACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGG
		ATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCA
		ACAAGGCCCTGCCTGCTCCTATCGAAAAGACCATCTCCAA
		GGCCAAGGGACAGCCCAGGGAACCCCAGGTTTACACCCT
		GCCTCCATGCCGGGAAGAGATGACCAAGAACCAGGTGTC
		CCTGTGGTGCCTGGTTAAGGGCTTCTACCCCTCCGATATCG
		CCGTGGAATGGGAGTCTAATGGCCAGCCTGAGAACAACT
		ACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCATT
		CTTCCTGTACTCCAAGCTGACAGTGGACAAGTCCAGATGG
		CAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGG
		CCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGAG
		CCCCGGCAAA
212	TPP-46960	GAGGTGCAGTTGGTCCAATCAGGAGCAGAAGTCAAAAAA
	LRRC15	CCCGGTGCTTCCGTTAAAGTATCTTGCAAAGCTAGCGGGT
	Heavy Chain	ACAAGTTCTCTTCCTATTGGATCGAGTGGGTTAAGCAGGC
	Hole (TCR)	ACCTGGACAGGGTCTTGAATGGATAGGCGAAATCCTTCCC
		GGTAGCGACACCACAAACTATGCTCAAAAGTTCCAAGATA
		GAGTTACTTTCACTTCAGATACTTCCATATCTACTGCATAT
		ATGGAACTGTCCCGTTTGAGGAGTGATGACACAGCAGTGT
		ACTACTGTGCCCGTGACCGTGGAAACTACCGTGCATGGTT
		CGGCTATTGGGGTCAGGGTACACTCGTAACAGTGAGCTCA
		GCAAGTACTAAGGGCCCCTCTGTGTTCCCTCTGGCTCCTTC
		CAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGCTGC
		CTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGTCCT
		GGAACTCTGGCGCTCTGACATCTGGCGTGCACACCTTTCC
		AGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCTCTG
		TCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGACCTA
		CATCTGCAATGTGAACCACAAGCCTTCCAACACCAAGGTG
		GACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACCGGC
		GGATCTGAGGTGCAGCTGGTGGAATCTGGCGGAGGATTG
		GTTCAGCCTGGCGGCTCTCTGAGACTGTCTTGTGTGGCTA
		GCGGCGACGTGCACAAGATCAACTTTCTCGGCTGGTACAG
		ACAGGCCCCTGGCAAAGAGAGAGAGAAGGTCGCCCACAT
		CTCCATCGGCGACCAGACCGATTACGCCGACTCTGCCAAG
		GGCAGATTCACCATCTCTCGGGACGAGTCCAAGAACATGG
		TGTACCTGCAGATGAACTCCCTGAAGCCTGAGGATACCGC
		CGTGTACTTCTGCCGGGCCTTCTCTCGGATCTACCCCTACG
		ATTATTGGGGCCAGGGCACCCTGGTCACAGTTTCTAGCGG
		CGGAGGCGGCTCTACCCATACCTGTCCTCCATGTCCTGCTC
		CTCCAGTGGCTGGCCCATCCGTGTTTCTGTTCCCTCCAAAG
		CCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAGTGA
		CCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCGAAGT
		GAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAA
		CGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCAC
		CTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGAT
		TGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAAC
		AAGGCCCTGCCTGCTCCTATCGAAAAGACCATCTCCAAGG
		CCAAGGGCCAGCCTCGGGAACCTCAAGTCTGTACCCTGCC
		TCCTAGCCGGGAAGAGATGACCAAGAACCAGGTGTCCCT
		GTCCTGTGCCGTGAAGGGCTTCTACCCTTCCGATATCGCC
		GTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTAC
		AAGACCACACCTCCTGTGCTGGACTCCGACGGCTCATTCT
		TCCTGGTGTCCAAGCTGACAGTGGACAAGTCCAGATGGCA
		GCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCC
		CTGCACAACAGGTTCACCCAGAAGTCCCTGTCTCTGAGCC
		CTGGCAAG
213	TPP-46960	GACATACAGATGACTCAGTCTCCATCTTCCCTTTCCGCTTC
	LRRC15	TGTGGGTGATCGAGTAACAATTACATGCCGGGCATCACAG
	Light Chain	GATATTTCTAACTATCTTAATTGGTATCAACAAAAACCAG
		GTAAGGCTCCAAAGTTTCTGATCTATTATACCTCTCGGCTG
		AACAGTGGTGTCCCCAGTAGGTTCAGCGGTAGTGGCAGCG
		GAACTGACTACACCCTGACCATTTCTTCTCTGCAACCCGA
		AGATTTTGCAACCTACTATTGTCAACAAGGCAACGCTCTT
		CCTTGGACCTTTGGCGGAGGCACAAAGGTCGAAATAAAA
		CGGACAGTGGCCGCACCGTCCGTGTTCATCTTCCCACCTTC
		CGACGAGCAGCTGAAGTCTGGCACAGCCTCTGTCGTGTGC
		CTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCAGT
		GGAAGGTGGACAATGCCCTGCAGTCCGGCAACTCCCAAG
		AGTCTGTGACCGAGCAGGACTCCAAGGACAGCACCTACA
		GCCTGTCCTCCACACTGACCCTGTCCAAGGCCGACTACGA
		GAAGCACAAGGTGTACGCCTGCGAAGTGACCCATCAGGG
		CCTGTCTAGCCCTGTGACCAAGTCTTTCAACCGGGGCGAG
		TGC
214	TPP-47826	GAGGTGCAGCTTGTTCAAAGCGGTGCAGAGGTTAAGAAG
	LRRC15	CCTGGAGCTAGCGTAAAAGTGTCCTGCAAAGCTAGTGGGT
	Heavy Chain	ATAAGTTCTCTAGCTACTGGATCGAGTGGGTGAAACAAGC
	Knob (CD8)	CCCAGGGCAGGGGTTGGAGTGGATTGGAGAAATACTCCC
		CGGATCAGACGCCACTAACTATGCACAGAAGTTTCAAGAC
		AGGGTTACCTTCACATCAGACACATCCATATCAACCGCAT
		ATATGGAGTTGTCCAGGCTGCGGAGCGACGACACCGCTGT
		CTATTATTGTGCCAGGGACCGGGGGAACTACAGGGCTTGG
		TTTGGGTACTGGGGACAGGGTACACTCGTCACAGTGTCCT
		CCGCAAGCACCAAGGGACCCTCTGTGTTCCCTCTGGCTCC
		TTCCAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGC
		TGCCTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGT
		CCTGGAACTCTGGCGCTCTGACATCTGGCGTGCACACCTT
		TCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCT
		CTGTCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGAC
		CTACATCTGCAATGTGAACCACAAGCCTTCCAACACCAAG
		GTGGACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACC
		GGCGGATCTGAGGTACAGCTGCTGGAATCAGGAGGCGGC
		TTGGTTCAGCCAGGAGGCTCACTTCGGCTCTCCTGCGCAG
		CAAGTGGGTTTACATTTACTGATTATGCTATAGGCTGGTTT
		CGGCAAGCACCTGGTAAGGAACGCGAAGGTGTGTCATGT
		ATTCGCGTAAGCGACGGATCAACATACTATGCAGACAGTG
		TCAAAGGTAGATTTACTATTAGTCGTGACAGCTCAAAGAA
		TACTCTGTACTTGCAAATGAATTCCCTTCGTGCCGAAGAT
		ACTGCCGTTTACTATTGCGCCGCAGGATCACTCTACACCT
		GCGTGCAGAGCATTGTTTGGCCTGCCAGGCCTTATTATGA
		TATGGACTATTGGGGCCAAGGCACTTTGGTCACTGTTAGC
		TCTGGAGGCGGAGGCAGCACCCATACCTGTCCTCCATGTC
		CTGCTCCTCCAGTGGCTGGCCCTTCCGTGTTTCTGTTCCCT
		CCAAAGCCTAAGGACACCCTGATGATCTCTCGGACCCCTG
		AAGTGACCTGCGTGGTGGTGGATGTGAAGCACGAGGATC
		CCGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGT
		GCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTACAA
		CTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCAC
		CAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTG
		TCCAACAAGGCCCTGCCTGCTCCTATCGAAAAGACCATCT
		CCAAGGCCAAGGGACAGCCCAGGGAACCCCAGGTTTACA
		CCCTGCCTCCATGCCGGGAAGAGATGACCAAGAACCAGG
		TGTCCCTGTGGTGCCTGGTTAAGGGCTTCTACCCCTCCGAT
		ATCGCCGTGGAATGGGAGTCTAATGGCCAGCCTGAGAAC
		AACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCT
		CATTCTTCCTGTACTCCAAGCTGACAGTGGACAAGTCCAG
		ATGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCAC
		GAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTC
		TGAGCCCCGGCAAA
215	TPP-47826	GAAGTGCAGTTGGTGCAGTCTGGGGCCGAAGTTAAGAAA
	LRRC15	CCAGGGGCCTCTGTTAAAGTAAGTTGCAAGGCTTCCGGTT
	Heavy Chain	ACAAGTTTTCAAGCTACTGGATCGAGTGGGTCAAGCAGGC
	Hole (TCR)	CCCAGGCCAAGGGTTGGAATGGATAGGAGAGATCCTTCC
		AGGGAGCGATGCCACAAACTACGCCCAAAAATTCCAGGA
		CAGGGTTACTTTTACTAGTGACACTTCCATATCAACAGCCT
		ATATGGAGCTGTCCCGATTGCGCTCCGATGATACTGCCGT
		GTACTATTGCGCCAGGGATAGAGGCAATTACCGCGCATGG
		TTTGGATACTGGGGGCAGGGAACACTTGTGACAGTTAGCT
		CCGCAAGCACCAAGGGCCCATCCGTCTTCCCCCTGGCACC
		CTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGC
		TGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGT
		CGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTT
		CCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGC
		AGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGA
		CCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAA
		GGTGGACAAGAGAGTTGAACCAAAAAGCTGTGATAAAAC
		CGGAGGTTCAGACGTGCAGTTGGTGGAATCTGGAGGCGGT
		GTTGTACAGCCCGGCGGCTCTTTGAGACTCTCATGTGTCG
		CCAGCGGCTACGTACACAAGATCAATTTTTATGGCTGGTA
		TAGGCAAGCTCCCGGTAAAGAGCGCGAGAAAGTCGCTCA
		TATTAGCATCGGAGATCAGACTGACTACGCCGACTCCGCT
		AAAGGGCGCTTCACCATATCTAGGGATGAATCAAAGAAC
		ACTGTGTATCTCCAGATGAACTCTCTCCGCCCAGAGGATA
		CTGCCGCTTACTATTGTCGTGCTCTTTCCCGTATATGGCCT
		TATGATTACTGGGGACAGGGGACATTGGTTACTGTGAGTT
		CTGGTGGCGGAGGATCTACCCACACCTGTCCTCCATGCCC
		TGCTCCACCCGTGGCCGGCCCTTCCGTGTTCCTGTTTCCTC
		CAAAGCCTAAGGACACCCTGATGATCTCTCGGACCCCTGA
		AGTGACCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCC
		GAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTG
		CACAACGCCAAGACCAAGCCTAGAGAGGAACAGTACAAC
		TCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACC
		AGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGT
		CCAACAAGGCCCTGCCTGCCCCCATCGAAAAGACCATCTC
		TAAGGCTAAGGGCCAGCCTCGCGAGCCTCAAGTCTGTACA
		CTGCCTCCTAGCCGGGAAGAGATGACCAAGAACCAGGTG
		TCCCTGTCTTGCGCCGTGAAGGGCTTCTACCCTTCCGATAT
		CGCCGTGGAATGGGAGTCCAATGGCCAGCCTGAGAACAA
		CTACAAGACAACCCCTCCTGTGCTGGACTCCGACGGCTCA
		TTCTTCCTGGTGTCCAAGCTGACAGTGGACAAGTCCAGAT
		GGCAGCAGGGCAACGTGTTCTCCTGCTCTGTGATGCACGA
		GGCCCTGCACAACCGGTTCACCCAGAAATCCCTGTCTCTG
		TCCCCTGGCAAG
216	TPP-47826	GATATTCAGATGACACAATCCCCTAGCAGTCTCAGTGCCT
	LRRC15	CAGTAGGGGACCGCGTCACAATAACATGTAGAGCCTCCCA
	Light Chain	GGATATTTCTAACTACTTGAACTGGTACCAGCAAAAGCCC
		GGTAAAGCCCCTAAATTCCTTATTTACTACACAAGCCGAT
		TGCACTCTGGTGTACCATCCAGGTTTAGTGGAAGCGGCTC
		TGGTACAGATTACACACTGACCATTTCATCCCTGCAACCA
		GAGGATTTCGCCACATATTACTGCCAACAGGGAAACGCCC
		TCCCTTGGACCTTTGGTGGGGGAACTAAAGTTGAGATCAA
		GCGGACAGTGGCCGCACCGTCCGTGTTCATCTTCCCACCT
		TCCGACGAGCAGCTGAAGTCTGGCACAGCCTCTGTCGTGT
		GCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCA
		GTGGAAGGTGGACAATGCCCTGCAGTCCGGCAACTCCCAA
		GAGTCTGTGACCGAGCAGGACTCCAAGGACAGCACCTAC
		AGCCTGTCCTCCACACTGACCCTGTCCAAGGCCGACTACG
		AGAAGCACAAGGTGTACGCCTGCGAAGTGACCCATCAGG
		GCCTGTCTAGCCCTGTGACCAAGTCTTTCAACCGGGGCGA
		GTGC
217	TPP-49058	GAGGTGCAGTTGGTCCAATCAGGAGCAGAAGTCAAAAAA
	LRRC15	CCCGGTGCTTCCGTTAAAGTATCTTGCAAAGCTAGCGGGT
	Heavy Chain	ACAAGTTCTCTTCCTATTGGATCGAGTGGGTTAAGCAGGC
	Knob (CD8)	ACCTGGACAGGGTCTTGAATGGATAGGCGAAATCCTTCCC
		GGTAGCGACACCACAAACTATGCTCAAAAGTTCCAAGATA
		GAGTTACTTTCACTTCAGATACTTCCATATCTACTGCATAT
		ATGGAACTGTCCCGTTTGAGGAGTGATGACACAGCAGTGT
		ACTACTGTGCCCGTGACCGTGGAAACTACCGTGCATGGTT
		CGGCTATTGGGGTCAGGGTACACTCGTAACAGTGAGCTCA
		GCAAGCACCAAGGGACCCTCTGTGTTCCCTCTGGCTCCTT
		CCAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGCTG
		CCTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGTCCT
		GGAACTCTGGCGCTCTGACATCTGGCGTGCACACCTTTCC
		AGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCTCTG
		TCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGACCTA
		CATCTGCAATGTGAACCACAAGCCTTCCAACACCAAGGTG
		GACAAGAGAGTGGAACCCAAGTCCTGCGATAAGACCGGC
		GGATCTGAGGTACAGCTGCTGGAATCAGGAGGCGGCTTG
		GTTCAGCCAGGAGGCTCACTTCGGCTCTCCTGCGCAGCAA
		GTGGGTTTACATTTACTGATTATGCTATAGGCTGGTTTCGG
		CAAGCACCTGGTAAGGAACGCGAAGGTGTGTCATGTATTC
		GCGTAAGCGACGGATCAACATACTATGCAGACAGTGTCA
		AAGGTAGATTTACTATTAGTCGTGACAGCTCAAAGAATAC
		TCTGTACTTGCAAATGAATTCCCTTCGTGCCGAAGATACT
		GCCGTTTACTATTGCGCCGCAGGATCACTCTACACCTGCG
		TGCAGAGCATTGTTTGGCCTGCCAGGCCTTATTATGATAT
		GGACTATTGGGGCCAAGGCACTTTGGTCACTGTTAGCTCT
		GGAGGCGGAGGCAGCACCCATACCTGTCCTCCATGTCCTG
		CTCCTCCAGTGGCTGGCCCTTCCGTGTTTCTGTTCCCTCCA
		AAGCCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAG
		TGACCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCGA
		AGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCAC
		AACGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCC
		ACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGG
		ATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCA
		ACAAGGCCCTGCCTGCTCCTATCGAAAAGACCATCTCCAA
		GGCCAAGGGACAGCCCAGGGAACCCCAGGTTTACACCCT
		GCCTCCATGCCGGGAAGAGATGACCAAGAACCAGGTGTC
		CCTGTGGTGCCTGGTTAAGGGCTTCTACCCCTCCGATATCG
		CCGTGGAATGGGAGTCTAATGGCCAGCCTGAGAACAACT
		ACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCATT
		CTTCCTGTACTCCAAGCTGACAGTGGACAAGTCCAGATGG
		CAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGG
		CCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGAG
		CCCCGGCAAA
218	TPP-49058	GAGGTGCAGTTGGTCCAATCAGGAGCAGAAGTCAAAAAA
	LRRC15	CCCGGTGCTTCCGTTAAAGTATCTTGCAAAGCTAGCGGGT
	Heavy Chain	ACAAGTTCTCTTCCTATTGGATCGAGTGGGTTAAGCAGGC
	Hole (TCR)	ACCTGGACAGGGTCTTGAATGGATAGGCGAAATCCTTCCC
		GGTAGCGACACCACAAACTATGCTCAAAAGTTCCAAGATA
		GAGTTACTTTCACTTCAGATACTTCCATATCTACTGCATAT
		ATGGAACTGTCCCGTTTGAGGAGTGATGACACAGCAGTGT
		ACTACTGTGCCCGTGACCGTGGAAACTACCGTGCATGGTT
		CGGCTATTGGGGTCAGGGTACACTCGTAACAGTGAGCTCA
		GCAAGCACCAAGGGACCCTCTGTGTTCCCTCTGGCTCCTT
		CCAGCAAGTCTACCTCTGGCGGAACAGCTGCTCTGGGCTG
		CCTGGTCAAGGACTACTTTCCTGAGCCTGTGACCGTGTCCT
		GGAACTCTGGCGCTCTGACATCTGGCGTGCACACCTTTCC
		AGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCTCTG
		TCGTGACCGTGCCTTCCAGCTCTCTGGGAACCCAGACCTA
		CATCTGCAATGTGAACCACAAGCCTTCCAACACCAAGGTG
		GACAAGAGAGTGGAACCAAAAAGCTGTGACAAGACTGGT
		GGCTCAGACGTACAATTGGTTGAAAGTGGTGGGGGCGTA
		GTCCAGCCCGGAGGCAGCCTCCGACTTTCCTGCGTCGCAT
		CTGGATACGTCCATAAAATCAACTTTTACGGCTGGTACAG
		ACAAGCACCAGGGAAAGAGCGGGAAAAAGTTGCCCACAT
		ATCCATTGGCGACCAGACAGATTATGCCGACAGCGCCAAG
		GGTCGATTTACTATAAGTCGGGACGAATCAAAGAATACTG
		TGTATCTCCAGATGAACAGTCTTCGACCTGAAGACACCGC
		CGCTTATTATTGTAGGGCTTTGTCCAGGATCTGGCCATACG
		ATTACTGGGGGCAAGGGACACTCGTAACCGTTAGTTCTGG
		TGGCGGAGGATCTACCCACACCTGTCCTCCATGCCCTGCT
		CCACCCGTGGCCGGCCCTTCCGTGTTCCTGTTTCCTCCAAA
		GCCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAGTG
		ACCTGCGTGGTGGTGGATGTGAAGCACGAGGATCCCGAA
		GTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACA
		ACGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCA
		CCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGA
		TTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAA
		CAAGGCCCTGCCTGCCCCCATCGAAAAGACCATCTCTAAG
		GCTAAGGGCCAGCCTCGCGAGCCTCAAGTCTGTACACTGC
		CTCCTAGCCGGGAAGAGATGACCAAGAACCAGGTGTCCCT
		GTCTTGCGCCGTGAAGGGCTTCTACCCTTCCGATATCGCC
		GTGGAATGGGAGTCCAATGGCCAGCCTGAGAACAACTAC
		AAGACAACCCCTCCTGTGCTGGACTCCGACGGCTCATTCT
		TCCTGGTGTCCAAGCTGACAGTGGACAAGTCCAGATGGCA
		GCAGGGCAACGTGTTCTCCTGCTCTGTGATGCACGAGGCC
		CTGCACAACCGGTTCACCCAGAAATCCCTGTCTCTGTCCC
		CTGGCAAG
219	TPP-49058	GACATCCAGATGACCCAGAGTCCCTCTAGTCTCTCAGCTT
	LRRC15	CCGTGGGGGACCGCGTCACTATAACTTGCCGTGCCTCACA
	Light Chain	AGACATATCAAACTACCTCAACTGGTATCAACAAAAGCCT
		GGAAAAGCTCCTAAGTTTTTGATTTATTACACTTCCAGGCT
		GGAGTCCGGTGTCCCATCCAGGTTCAGCGGCAGTGGAAGC
		GGAACCGACTATACACTGACTATTTCTAGCTTGCAACCCG
		AGGACTTTGCCACCTATTATTGCCAGCAAGGTAACGCCCT
		GCCCTGGACCTTCGGTGGCGGAACCAAGGTGGAAATCAA
		GCGGACAGTGGCCGCACCGTCCGTGTTCATCTTCCCACCT
		TCCGACGAGCAGCTGAAGTCTGGCACAGCCTCTGTCGTGT
		GCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCA
		GTGGAAGGTGGACAATGCCCTGCAGTCCGGCAACTCCCAA
		GAGTCTGTGACCGAGCAGGACTCCAAGGACAGCACCTAC
		AGCCTGTCCTCCACACTGACCCTGTCCAAGGCCGACTACG
		AGAAGCACAAGGTGTACGCCTGCGAAGTGACCCATCAGG
		GCCTGTCTAGCCCTGTGACCAAGTCTTTCAACCGGGGCGA
		GTGC
220	CH1 with	ASTKGPSVFPLAPSSKSTSGGTADLGCLVKDYFPEPVTVSW
	A141D	NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
		NVNHKPSNTKVDKRVEPKSC
221	IgG1 CH2	APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
	WT(IMGT)	KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
	alternate	WLNGKEYKCKVSNKALPAPIEKTISKAK
222	IgG1 CH2 Fc-	APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV
	null (IMGT)	KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD
	alternate	WLNGKEYKCKVSNKALPAPIEKTISKAK

Example 2: In Vitro Testing of LM1486 Binding Proteins

The activity of the novel LM1486 (CD20) binding proteins disclosed herein were demonstrated.

In Vitro Cytolysis

Method

An assay was conducted to measure cytolysis of LM1486 compared to known CD3 engagers. The assay was run for 4 days with an OCI-Ly18 CD20+ DLBCL line and PBMCs at an E:T ratio of 5:1. Target B cell survival was assessed by flow cytometry.

Results

FIGS. 1A and 1B show potent in vitro cytolysis by LM1486 compared to conventional CD3 engagers. FIG. 1A shows the in vitro cytolytic activity of LM1486 and CD3 engagers with monovalent or bivalent engagement to CD20. FIG. 1B shows the corresponding EC₅₀s in the cytolysis in vitro assay.

Comparison of CD4 T Cell and CD8 T Cell Activation with Known CD3 Engager

Method

At the end of the cytolysis assay, T cell activation status was evaluated by flow cytometry.

Results

A CD8-biased T cell activation profile was induced by LM1486 compared to conventional CD3 engagers. FIG. 1C shows corresponding CD4 T cell activation profiles as % of cells expressing CD25 on the cell surface. FIG. 1D shows corresponding CD8 T cell activation profiles as % of cells expressing CD25.

Comparison of Cytokine Release Profiles with known CD3 Engagers

Method

The cytokine release profiles were measured in the supernatant of an in vitro cytolysis experiment using a multiplexing assay.

Results

FIGS. 2A-2D show in vitro cytokine release profiles of LM1486 compared to conventional CD3 engagers: IL-6 (FIG. 2A), IL-10 (FIG. 2B), TNF-α (FIG. 2C), and IL-17A (FIG. 2D). The results show that LM1486 induced much lower levels of cytokines when compared with the CD3×CD20 bivalent engager (FIG. 26A) and levels similar to the CD3×CD20 monovalent engager.

In Vitro Cytolysis in Different CD20+ B Cell Lines

Method

The in vitro cytolytic activity of LM1486 was measured with different CD20+ B cell lines incubated with PBMCs at an E:T ratio of 5:1. The assay was run for 3 to 4 days and target B cell survival was assessed by flow cytometry.

Results

FIG. 3 shows potent in vitro cytolytic activity of LM1486 across an array of B cell lines expressing various levels of CD20 antigen density and also shows the absence of correlation between CD20 density and EC₅₀s.

Comparison of Cytolytic Activity of LM1486 with CD8 and TCR Null Controls

Method

LM1486 was compared with control binding proteins TENG0501 (TCR null) and TENG0502 (CD8 null). The assay was run with an OCI-Ly18 CD20+ DLBCL line and PBMCs at an E:T ratio of 5:1, and target B cell survival and T cell activation profiles were assessed by flow cytometry.

FIGS. 4A-4C illustrate the structures of LM1486 (FIG. 4A) and the structures of modified binding proteins with a null binding domain replacing the anti-CD8 domain (FIG. 4B) or a null binding domain replacing the anti-TCR binding domain (FIG. 4C).

Results

FIG. 4D shows the in vitro cytolytic activity of LM1486 and control binding proteins TENG0501 and TENG0502. FIG. 4E shows corresponding T cell activation profiles as % CD4 and CD8 T cells expressing CD25 assessed by flow cytometry. The data demonstrates that the LM1486 TCR binding domain is essential to cytolytic activity and that CD8 arm enhances potency and confers CD8-biased T cell activation.

Comparison of Cytolytic Activity of LM1486 with Single Null Binding Domain Controls

Method

LM1486 was compared with control binding proteins TENG0500 (null binding protein replacing anti-CD20 domain on anti-CD8 arm) and TENG0499 (null binding protein replacing anti-CD20 domain on anti-TCR arm). The assay was run with OCI-Ly18 CD20+ DLBCL line and PBMCs at an E:T ratio of 5:1, and target B cell killing and T cell activation profiles were assessed by flow cytometry.

FIGS. 5A-5C illustrate the structure of LM1486 (FIG. 5A) and the structure of modified binding proteins with a null binding domain replacing the anti-CD20 domain on the anti-CD8 arm (FIG. 5B) or with a null binding domain replacing the anti-CD20 binding domain on the anti-TCR arm (FIG. 5C).

Results

FIG. 5D shows the in vitro cytolytic activity of LM1486 and control binding proteins TENG0499 and TENG0500. FIG. 5E shows corresponding T cell activation profiles as % CD4 and CD8 T cells expressing CD25. These data demonstrate that CD20 binding on the TCR arm is required to achieve cytolysis.

Comparison of Cytolytic Activity of LM1486 with LM1486-2

LM1486 differs from LM1486-2 by the presence of an RF mutation in the TCR heavy chain of LM1486 (see SEQ ID NO: 110), H318R and Y319F.

Method

The assay was run for 3 days with OCI-Ly18 CD20+ DLBCL line and PBMCs at an E:T ratio of 5:1. B cell survival and T cell activation profiles were assessed by flow cytometry.

Results

FIG. 9 shows overlapping in vitro cytolysis profiles for LM1486 and LM1486-2. FIGS. 9A and 9D show cytotoxicity profiles in a range of T cell engager concentrations for 2 different PBMC donors. FIGS. 9B, 9C, and FIG. 9E, 9F show associated CD4 and CD8 T cell activation profiles as % of CD25 surface expression for 2 donors, respectively.

Example 3: In Vivo Efficacy of LM1486 in B Cell Model

Assessing Anti-Tumor Efficacy In Vivo in a Disseminated B Cell Model

Method

At day 6, animals were humanized with 5e6 PBMCs. Animals were then treated i.p. twice a week with PBS, or LM1486 at 0.8, 0.08, or 0.008 mg/kg. Tumor burden was monitored twice weekly by bioluminescence imaging and total flux (p/s) was recorded.

Results

The results demonstrate that the LM1486 binding protein was efficacious in vivo. FIG. 6 shows tumor burden in a disseminated model of the PBS treated groups (FIG. 6A), groups treated LM1486 at 0.8 mg/kg (FIG. 6B), 0.08 mg/kg (FIG. 6C), and 0.008 mg/kg (FIG. 6D). These data demonstrate that LM1486 confers potent anti-tumor efficacy in vivo with as low as 0.008 mg/kg.

Comparing Tumor Growth Inhibition, Survival Benefit, and Superior Safety Over Known CD3×CD20 Known Engagers

Method

CD34+ engrafted NSG mice (n=8-9/group) were implanted s.c. on the flank with 5e6 OCI-Ly18 cells. Animals were then treated i.p. with 1 mg/kg of T cell engagers (CD3×CD20 monovalent (FIG. 26B) or bivalent engager (FIG. 26A) or LM1486 or PBS) at day 1, followed by weekly engager dosing until day 21. Tumor growth was monitored twice weekly until study end. Animals were euthanized when tumor reached 2000 mm³. Percentage survival is reported for all treatment groups up to study end (day 42), weight loss was monitored over time as a surrogate assessment of safety.

Results

FIG. 7 shows LM1486 potent tumor growth inhibition, survival benefit, and superior safety over known CD3 engagers in a fully humanized in vivo model with subcutaneous B cell tumor. FIGS. 7A-7C shows LM1486 potent tumor growth inhibition, survival benefit, and superior safety over conventional CD3 engagers in a fully humanized in vivo model with subcutaneous B cell tumor. FIG. 7C shows the percentage body weigh loss recorded 72h post first dose of T cell engager.

Anti-Tumor Efficacy of LM1486 in a Disseminated DLBCL In Vivo Model Compared to Known CD3×CD20 Engagers

Method

NSG mice were injected i.v. with Toledo B cell line (DLBCL origin) expressing Luciferase (Toledo-Luc) at day 0. At day 7, mice were humanized with 10e6 PBMCs. At day 21, a single dose of T cell engager was injected at i.v at 1 mg/kg for LM1486 and equimolar concentrations for the other molecules. At day 24, tumor burden was evaluated using bioluminescence imaging (BLI). Total flux (p/s) reflecting tumor burden are displayed for each treatment group. 3h post engager treatment, animals were bled, and levels of circulating cytokines were evaluated using a multiplexing assay.

Results

FIGS. 8B-E show reduced cytokine release: TNF-α (pg/mL) (FIG. 8B), IL-10 (pg/mL) (FIG. 8C), IL-2 (pg/mL) (FIG. 8D) and IL-17A (pg/mL) (FIG. 8E) compared to a known CD3×CD20 bivalent engager (FIG. 26A). Further, FIG. 8F shows reduced weight loss compared to the same known CD3×CD20 bivalent engager. LM1486, in a disseminated DLBCL in vivo model, has a superior safety profile compared to a known CD3×CD20 bivalent engager (FIG. 26A) (reduced systemic cytokine and reduced weight loss).

Example 4: In Vitro Testing of LRC150016 Binding Proteins

A novel T-cell engager, LRC150016 (LRRC15), was generated, featuring two LRRC15-binding domains, a CD8 binding domain and a TCR binding domain (FIG. 15).

Results

Binding Protein Structure and Cell Binding Activity.

LRC150016 was assessed for binding onto LRRC15^Pos and LRRC15^neg cell lines. LRC150016 engager shows specific binding to LRRC15 expressing cell lines. The assay was performed by incubating LRC150016 with LRRC15^pos cell lines (FIG. 10A) Saos-2 WT (FIG. 10B) and RPMI WT (FIG. 10C)), LRRC15^neg cell lines (A431, Saos-2 KO and RPMI-7951 KO), for 1h at 4° C. LRC150016 was detected using anti-human Fc Alexa Fluor 647 conjugated secondary antibody (MFI). LRC150016 is able to bind to LRRC15^pos tumour cell lines Saos-2 and RPMI7951, but not to the LRRC15^neg cell line A431 (FIG. 10A). When LRRC15 is knocked out from Saos2 WT and from RMPI-7951 WT cells, the binding of LRC150016 is lost, demonstrating that the observed binding is specifically due to LRRC15 protein binding (FIG. 10B and FIG. 10C).

LRC150016 Mediates T-Cell Effector Functions in an Antigen (LRRC15) Dependent Manner.

LRC150016 was assessed for in-vitro T-cell effector functions on Saos-2 WT(LRRC15⁺) and Saos-2 KO(LRRC15⁻) target cell lines. Purified human T cells were co-cultured with target cell lines in the presence of LRC150016 at an E:T ratio of 4:1 for 3 days. % of target cytolysis (cytotoxicity) was measured by CELL-TITRE-GLO. Cytokines (TNFα, IFNγ and IL-6) were measured in culture medium collected post 3 days of co-culture by a multiplexing cytokine assay. FIG. 11A depicts that LRC150016 induces a dose-dependent strong cytolysis of Saos-2 WT (LRRC15⁺), but not Saos-2 KO (LRRC15⁻) target cells. FIG. 11B shows cytokines associated with the induction of T-cell effector function mediated by LRC150016 upon co-culture of T-cells with Saos-2 WT cells, but not with LRRC15-KO Saos-2 cells. FIG. 11A-11B shows LRC150016 engager mediates T-cell effector functions in an antigen (LRRC15) dependent manner.

LRC150016 Specifically Induces Activation of CD8 T-Cells, Over CD4 T-Cells, Against LRRC15⁺ Saos-2 Cells.

To compare LRC150016 with a Comparator bispecific LRRC15/CD3 T-cell engager, whole healthy human PBMCs (effectors) were co-cultured with Saos-2 WT (LRRC15+) target cells in the presence of molecules for a period of 4 days at an E:T ratio 4:1 (FIG. 12A). Post 4 days co-culture, target cell cytolysis, activation of T-cells and secreted cytokines were measured by CELL-TITRE-GLO, CD25 surface expression assessment by flow cytometry and a multiplex cytokine assay respectively. Activation of CD8 T-cells in this co-culture, measured by upregulation of CD25 is similar for both molecules (FIG. 12B), but the CD3-targeted bispecific also strongly induces unwanted activation of CD4 T-cells, also shown by CD25 upregulation (FIG. 12C). FIG. 12 demonstrates potent in vitro cytolysis and CD8-biased T-cell activation profile induced by LRC150016, compared to Comparator CD3 TCE.

Despite similar target cell cytolysis activity, LRC150016 induced lower levels of cytokines, compared to the CD3-targeting Comparator TCE (FIGS. 13A-D). FIGS. 13A-13D show the in vitro cytokine profile of LRC150016, compared to the Comparator CD3 TCE. Cytokine profiles were measured in the supernatant of whole PBMC in vitro cytolysis experiments using a multiplexing cytokine assay. TNFα (FIG. 13A), IL-6 (FIG. 13B), IL-10 (FIG. 13C), and IL-2 (FIG. 13D).

LRC150016 Mediates T-Cell Cytolytic Activity Against LRRC15⁺ Cells Across a Range of LRRC15 Expression Levels.

To understand the correlation between LRRC15 copy number (density) on target cells and LRC150016 mediated target cell cytolysis, purified human T cells were co-cultured with LRRC15^pos target cell lines, from various indications, in the presence of LRC150016 for a period of 6-days at an E:T ratio of 4:1. Target cell cytolysis was measured by CELL-TITRE-GLO. E_max and EC₅₀ values were derived from these experiments and projected against LRRC15 copy number of corresponding target cells. In co-cultures of tumour cells with isolated CD3 T-cells, LRC150016 induces tumour cell cytolysis of cells with more than 10-fold variation in LRRC15 antigen copy number (FIG. 14). FIG. 14A and FIG. 14B demonstrates that LRC150016 shows specific in vitro cytolytic activity across an array of cell lines expressing various levels of LRRC15.

Example 5: In Vitro and In Vivo Testing of LM1653 (GPC3) Binding Proteins

Binding Kinetics Assay

Method

Binding kinetics were measured by biolayer interferometry on an Octet384 instrument. Streptavidin (SA) biosensors were loaded with biotinylated protein antigens (ACRO Biosystems, Newark, DE) in PBS pH 7.2, 1 mg/mL BSA, 0.05% (v/v) TWEEN (Kinetic buffer). The loaded biosensors were washed in the same buffer before carrying out association and dissociation measurements with various antibodies for the indicated times. Kinetic parameters (K_on and K_off) and affinities (K_D) were calculated from a non-linear fit of the data using the OCTET software v.12.2.1.24.

Cell Binding Assay

Method

Cell bindings were measured by flow cytometry. Human hepatoma cells HepG2 and Hep3B, human and cyno PBMC were incubated with serial diluted GPC3 TITAN molecule on ice for 30 minutes, washed with FACS buffer and stained with goat anti-human-IgG-APC (Jackson ImmunoResearch) on ice for 30 minutes followed by flow cytometry analysis.

Results

FIG. 25 shows LM1653 binds human hepatoma and effector cells. FIG. 25 shows that GPC3 TITAN LM1653 binds human hepatoma cells Hep3B at EC₅₀: 8 nM (FIG. 25A) and HepG2 at EC₅₀: 2 nM (FIG. 25B). GPC3 TITAN binds to human PBMC at EC₅₀: 6 nM (FIG. 25C). Cell bindings were measured by flow cytometry. Human hepatoma cells HepG2 and Hep3B, human PBMC were incubated with serial diluted GPC3 TITAN molecule on ice for 30 minutes, washed with FACS buffer and stained with goat anti-human-IgG-APC (Jackson ImmunoResearch) on ice for 30 minutes followed by flow cytometry analysis.

TABLE 2
Kinetic analysis of anti-GPC3 Fab on GPC3 protein
	GPC3	Ka (1/Ms)	Kdis (1/s)	KD (M)
	human	2.26E05	4.22E−04	1.87E−09

TABLE 3
Kinetic analysis of anti-CD8 VHH on CD8α protein
	CD8α	Ka (1/Ms)	Kdis (1/s)	KD (M)
	Human	5.17E05	2.86E−02	5.53E−08

Table 2 shows kinetic analysis of anti-GPC3 fab on soluble recombinant human GPC3 protein interaction. Table 3 shows kinetic analysis of anti-CD8 VHH on soluble recombinant human CD8a protein interaction via Biolayer Interferomerty on an OCTET RED384.

Binding kinetics were measured by biolayer interferometry on an OCTET 384 instrument. Streptavidin (SA) biosensors were loaded with biotinylated protein antigens (ACRO Biosystems, Newark, DE) in PBS pH 7.2, 1 mg/mL BSA, 0.05% (v/v) TWEEN (Kinetic buffer). The loaded biosensors were washed in the same buffer before carrying out association and dissociation measurements with various antibodies for the indicated times. Kinetic parameters (K_on and K_off) and affinities (K_D) were calculated from a non-linear fit of the data using the OCTET software v.12.2.1.24.

In Vitro Cytolysis Induced by LM1653 Compared to a Conventional GPC3×CD3 Engager, Against Hepatocellular Carcinoma (HCC) Cell Lines

Method

GPC3 expression was detected with a commercially available anti-human GPC3 antibody and quantified using the BD Quantibrite kit. Cytotoxicity assay against the indicated HCC cell line was run for 4 days using PBMCs at an E:T ratio of 10:1, using the XCELLIGENCE system.

Results

FIG. 16 shows in vitro cytolysis induced by LM1653 compared to a conventional GPC3×CD3 engager, against hepatocellular carcinoma (HCC) cell lines expressing different levels of GPC3. FIG. 16A shows surface expression of GPC3 across 4 HCC cell lines. GPC3 expression was detected with a commercially available anti-human GPC3 antibody and quantified using the BD Quantibrite kit. FIG. 16B-E shows the in vitro cytolytic activity of LM1653 and of a monovalent GPC3×CD3 engager against the HCC cell lines HepG2 (FIG. 16B), Hep3B (FIG. 16C), Huh-7 (FIG. 16D), and PLC/PRF/5 (FIG. 16E). Graphs show mean±SEM (n=3-6 PBMC donors). The tables in the Figures show the corresponding EC₅₀ and killing E_max for each cell line.

T-Cell Activation Profiles

Method

T cell activation profiles were determined by flow cytometry. Activation profile was defined as % of parental cells expressing CD25.

Results

FIG. 17 shows CD8-biased T cell activation profile induced by LM1653 compared to a conventional GPC3×CD3 engager, against HCC cell line expressing different levels of GPC3. FIG. 17 shows CD8+ T cell (upper panels) and CD4+ T cell (bottom panels) activation profiles induced by LM1653 and by a monovalent GPC3×CD3 engager in cytotoxicity assay against HCC cell lines HepG2 (FIG. 17A), Hep3B (FIG. 17B), Huh-7 (FIG. 17C), and PLC/PRF/5 (FIG. 17D). Graphs show mean±SEM (n=3-6 PBMC donors). The tables show the EC₅₀ for CD8 and CD4 T cell activation for each cell line.

Comparison of Cytokine Release Profiles with a Known CD3 Engager

Method

Cytokine released in supernatant were measured with a multiplex assay (Luminex).

Results

FIG. 18 shows in vitro cytokine release profiles of LM1653 compared to a conventional GPC3×CD3 engager. FIG. 18 shows levels of secreted cytokines, TNF-α (FIG. 18A), IL-10 (FIG. 18B), IL-2 (FIG. 18C), and IFN-7 (FIG. 18D), in an in vitro cytolysis assay, using HCC Hep3B cell line as target cells at E:T ratio of 10:1. Results from one representative PBMC donor of 3 are shown.

T Cell Activation and Cytokine Release Profile Induced by LM1653 Compared to a Conventional GPC3×CD3 Engager

Method

Donor PBMCs (n=3) were incubated with LM1653 and a conventional GPC3×CD3 engager for 48 hours. CD8 and CD4 T cell activation profiles (defined as % of parental cells expressing CD25) were determined by flow cytometry. Cytokines released in supernatants at 48 hours were measured using a multiplex assay (LUMINEX).

Results

FIG. 19 shows non-specific T cell activation and cytokine release profile induced by LM1653 compared to a conventional GPC3×CD3 engager. FIG. 19 shows non-specific activity of LM1653 and of a conventional GPC3×CD3 engager in a plate-bound assay. Donor PBMCs (n=3) were incubated with LM1653 and a conventional GPC3×CD3 engager for 48 hours. CD8 and CD4 T cell activation profiles (defined as % of parental cells expressing CD25) are depicted in FIG. 19A and FIG. 19B, respectively. Histograms represent mean±SEM; *, p<0.05, 2-way ANOVA. Released IL-6 (FIG. 19C), TNF-α (FIG. 19D), IFN-7 (FIG. 19E), IL-2 (FIG. 19F), IL-10 (FIG. 19G), and IL-17 (FIG. 19H) were measured in the supernatant at 48 hours, using a multiplex assay (LUMINEX). Results from one representative donor PBMC are shown.

In Vitro Cytolysis and Cytokine Release Profile Induced by LM1653 and by a Conventional GPC3×CD3 Engager

Method

The assay was run for 4 days using either purified CD4 or CD8 T cells at an E:T ratio of 5:1, using the XCELLIGENCE systems. Target cells were HCC GPC3⁺ Hep3B. Cytokine released in supernatants were measured with a multiplex assay (LUMINEX).

Results

FIG. 20 shows in vitro cytolysis and cytokine release profile induced by LM1653 and by a conventional GPC3×CD3 engager, using purified CD4 and CD8 T cells. FIG. 20 shows the in vitro cytolytic activity (FIG. 20A-B) and cytokine secretion (FIG. 20C-J) of LM1653 and of a monovalent GPC3×CD3 engager using purified CD4 T cells (upper panels) and purified CD8 T cells (bottom panels), in a XCELLIGENCE assay.

In Vitro Cytolysis Induced by LM1653 Against GPC3⁺ Hep3B

Method

Dissociated tumor cells from HCC patients were used as source of effector cells in this assay, using the XCELLIGENCE systems. Tumor-infiltrating T cells were added to the culture at E:T ratio of 2:1 and 3:1, respectively. Target cells were HCC GPC3+ Hep3B.

Results

FIG. 21 shows in vitro cytolysis induced by LM1653 against GPC3⁺ Hep3B, using HCC patient-derived tumor infiltrating lymphocytes. FIG. 21 shows LM1653-dependent killing of GPC3⁺ Hep3B cell line by patients-derived tumor infiltrating lymphocytes obtained from HCC stage II (FIG. 21A) and stage IIIB (FIG. 21B). Tumor-infiltrating T cells were added to the culture at E:T ratio of 2:1 and 3:1, respectively. Cytotoxicity activity is shown for LM1653 and a null control compound unable to bind GPC3.

In Vitro Cytolysis Induced by LM1653 Against GPC3+ Lung and Ovarian Cancer Cells

Method

Cytotoxicity assays against the indicated ovarian and lung cell lines were run for 4 days using PBMCs at an E:T ratio of 10:1, using the XCELLIGENCE system.

Results

FIG. 22 shows in vitro cytolysis induced by LM1653 compared to a conventional GPC3×CD3 engager, against GPC3+ lung and ovarian cancer cell. FIG. 22 shows the in vitro cytolytic activity of LM1653 and of a monovalent GPC3×CD3 engager against non small cell lung cancer lines NCI-H661 (FIG. 22A) and NCI-H2172 (FIG. 22B) and ovarian cancer cell lines OV-90 (FIG. 22C) and Kuramochi (FIG. 22D). The tables in the figures show the corresponding EC₅₀ and killing E_max for each cell line

In Vitro Cytolysis and T Cell Activation Profile Induced by LM1653 and LM1653-2.

Method

Cytotoxicity assays against the indicated HCC cell line were run for 4 days using and cytolysis activity was measured at day 4, using PBMC at the ratio of 10:1 in the XCELLIGENCE system (n=1 PBMC donor). T cell activation profile was determined in cytotoxicity assays by flow cytometry.

Results

FIG. 23 shows comparable in vitro cytolysis and T cell activation profile induced by LM1653 and LM1653-2 (same amino acid sequence as LM1653, but produced in a different lot). FIG. 23A-B The tables show the corresponding EC₅₀ and killing E_max for GPC3+ cell line Hep3B (FIG. 23A) and Huh-7 (FIG. 23B). T cell activation profile was determined in cytotoxicity assay, using the Hep3B (FIG. 23C-D) and Huh-7 cell line (FIG. 23E-F). The percentage of CD8 (filled symbol) and CD4 (empty symbol) T cells expressing CD25 is reported.

In Vivo Anti-Tumor Activity of LM1653 Against GPC3^high and GPC3^low HCC Xenograft Models.

Method

GPC3^high

At day 0, NSG mice (n=10/group) were implanted SC with 5e06 Hep3B cells, expressing high levels of GPC3. On day 21, animals were humanized with 10e06 panT cells expanded in vitro. Animals were treated starting on day 22 IP once a week with PBS, LM1653 (1, 0.3, 0.1 and 0.03 mg/kg) and conventional GPC3×CD3 engager (1 and 0.1 mg/kg), for a total of 4 injections. Tumor burden was measured overtime and reported as mean±SEM. $, 40% tumor-free; **, p<0.01 and ***, p<0.001, 2-way Anova (FIG. 24A).

GPC3^low

At day 0, NSG mice (n=10/group) were implanted SC with 5e06 PLC/PRF/5 cells, expressing low levels of GPC3. On day 5, animals were humanized with 10e06 panT cells expanded in vitro. Animals were treated starting on day 6 IP twice a week with PBS and LM1653 (3, 1, and 0.3 mg/kg), for a total of 7 injections. Tumor burden was measured overtime and reported as mean±SEM. **, p<0.0001, 2-way Anova (FIG. 24B).

Results

FIG. 24 shows that LM1653 confers anti-tumor efficacy in vivo against GPC3^high and GPC3^low HCC xenograft models. Dotted lines represent TCE administrations. In vivo anti-tumour activity and associated cytokine secretion of LM1653 in Hep3B tumour bearing mice

Method

NSG mice (n=11-12 for each treatment group) were irradiated with 200 cGy at Day −3 and implanted IP with 7×10⁶ Hep3B-Luciferase cells at Day 0. At Day 8, mice were engrafted IV with 1×10⁷ human PBMCs from one healthy donor. At Day 22, mice were injected IP with either vehicle (PBS), LM1653-2 or GPC3×CD3 at 1 mg/kg. Tumor burden was monitored twice a week by bioluminescence imaging (BLI) until Day 24, 2 days post treatment.

Results

FIG. 41 shows that LM1653 induced significant tumor growth inhibition in Hep3B tumor bearing mice without significant associated cytokine secretion, in comparison to a conventional GPC3×CD3 engager.

FIG. 41A shows the tumor growth inhibition induced by both LM1653-2 and a conventional GPC3×CD3 engager. The mean of tumor burden+/−SEM across the course of the experiment in each treatment group is shown. Dashed line indicate day of treatment. The statistical significance of the differences in tumor burden overtime was determined using 2-way ANOVA with Tukey's multiple comparisons test (****p<0.0001; ns=not significant).

In FIG. 41B-E, serum cytokine concentrations were evaluated at 6 hours post TCE administration by multiplex assays. Individual values as well as mean±SEM for each treatment group is plotted for IFN-7 (FIG. 41B), TNF-α (FIG. 41C), IL-10 (FIG. 41D), and IL-2 (FIG. 41E). The statistical significance of the differences was determined using One-way ANOVA with Tukey's multiple comparisons test.

In Vivo Tumour Growth Inhibition Activity and Associated Cytokine Secretion of LM1653-3 in NCI-H661 Lung Tumor Bearing Mice

Methods

NSG mice (n=5-7 for each treatment group) were irradiated with 200 cGy at Day −3 and implanted IV with 2.5×10⁶ NCI-H661-Luciferase cells at Day 0. At Day 12, mice were engrafted IV with 1×10⁷ human PBMCs from one healthy donor. Starting at Day 27, mice were injected IV with either vehicle (PBS) or LM1653-3 at 1 and 0.3 mg/kg 2QW, for a total of 4 doses. OKT3 was used as positive control for T cell activation and cytokine secretion. LM1653-3 has the same amino acid sequence as LM1653 but was produced in a different lot.

Results

FIG. 42A shows that tumor growth inhibition was induced at both doses of LM1653-3.

FIG. 42 shows that LM1653-3 induced significant tumor growth inhibition in NCI-H661 lung tumor bearing mice without significant associated cytokine secretion, in comparison to vehicle-treated animals. Tumor burden was monitored twice a week by bioluminescence imaging (BLI). The geometric mean of tumor burden+/−SD across the course of the experiment in each treatment group is shown. Dashed line indicate days of treatment. The statistical significance of the differences in tumor burden overtime was determined using 2-way ANOVA with Tukey's multiple comparisons test (*, p<0.05; **p<0.01; ****p<0.0001). LM1653-3 exhibits the same in vitro bioactivity profile of LM1653 and LM1653-2.

In FIG. 42B-E, serum cytokine concentrations were evaluated at 3 hours post 1^st TCE or OKT3 administration by multiplex assays. Individual values as well as mean±SEM for each treatment group is plotted for IFN-7 (FIG. 42B), TNF-α (FIG. 42C), IL-10 (FIG. 42D), and IL-2 (FIG. 42E).

Example 6: Efficacy of LM1653 (GPC3 TITAN) in Cynomolgous Monkey Model

Method

Kinetic measurements to soluble forms of GPC3 and CD8ab were performed using a BIACORE instrument. K_D were calculated as the ratio of k_off/k_on from a non-linear fit of the data. Values are presented as mean±standard deviation (SD) of three experiments. Owing to assay limitations, kinetic values for TCR binding were unable to be determined accurately.

PBMC from 3 different healthy human or 4 cynomolgus monkey donors were incubated with HCC GPC3+ HepG2 at an effector to target ratio of 10:1. LM1653 was added in a 5-fold serial dilution starting from 50 nM for a total of 9 dilution doses. The percentage of cytolysis was assessed using XCELLIGENCE impedance-based RTCA MP analyzer at 48 hours post PBMC addition.

Cynomolgus monkey (n=3 per treatment group and per sex) were administered LM1653 weekly by intravenous (bolus) or subcutaneous injection for 1-month (5 doses), at doses of 1 mg/kg/week IV, 6 mg/kg/week IV or 6 mg/kg/week SC followed by a 2-week recovery period. Animals that received vehicle only as both IV & SC were used as controls.

Results

Table 4 shows the binding kinetics of LM1653 to recombinant human and cynomolgus monkey GPC3 and CD8 Proteins.

TABLE 4
Antigen	Kon (M−1s−1)	Koff (s−1)	KD (nM)
Human GPC3	3.41 ± 0.29 E+05	2.26 ± 0.04 E−04	0.67 ± 0.05
Cynomolgus	3.26 ± 0.04 E+05	2.67 ± 0.20 E−04	0.82 ± 0.06
monkey
GPC3
Human CD8ab	3.40 ± 0.23 E+05	2.80 ± 0.18 E−02	82.7 ± 11.0
Cynomolgus	5.27 ± 0.92 E+05	3.29 ± 0.59 E−02	62.4 ± 4.1
monkey
CD8ab

Table 5 shows LM1653-Induced T cell dependent cellular cytotoxicity (TDCC) in HCC HepG2 Cell Line, using human and cynomolgus Monkey PBMCs. Table 5 shows TDCC EC₅₀ and E_max median with interquartile range (IQR) for both human and cynomolgus monkey PBMCs.

	TABLE 5
	T cell Dependent Cellular Cytotoxicity
	Median EC50 (pM)
Species	(IQR = Q3 − Q1)	Emax (%)
Human (n = 3)	8.28	(23.01)	89.72	(14.27)
Cynomolgus monkey (n = 4)	31.55	(85.54)	67.36	(24.60)
Fold differences (c vs h)	3.8	0.76

FIG. 43 shows the effect of LM1653 administration in cynomolgus monkey. FIG. 43A shows the absolute number of CD8 T cells in blood over the course of treatment. Administration of LM1653 was associated with a transient decline in the number of CD8 T cells in the blood, as a result of T cell margination. Margination is considered an expected effect of LM1653 activity, as consequence of drug interaction with target T cells.

FIGS. 43B-E show serum cytokine and chemokine levels following the first dose administration. LM1653 was associated with a transient increase in the chemokines MCP-1 and MIP-1b, that are associated with CD8 T cell migration and function. Only marginal and transient increases were observed for CRS-associated cytokines IL-10 and IL-6, in agreement with the preclinical data generated in vitro and in mice. LM1653 activity is accompanied by only minimal increases in circulating cytokine level, potentially lowering the risk of CRS-associated toxicity in patients. Solid and dotted lines represent values obtained for male and female, respectively, and vehicle control.

Example 7: Humanisation of Anti-TCR VHH Framework Residues

Anti-TCR VHH amino acid sequence of SEQ ID NO: 42 is of camelid nature. Humanization of the amino acid residues to minimize immunogenicity was performed, where non-vernier framework residues were altered to match closest human germline sequence IGHV3-23*1. During humanization, the nanobody hallmark residues Y42, and K52 in framework 2 were not altered. The lead humanized VHH panel were generated where the hallmark residues E49 and R50 were either humanized or not humanized. A total of approximately 40 humanized variants were assessed with a 90% binding retention threshold set. Prior to introduction in trispecific antibody format, humanized clones were tested for binding retention by generation as Fc fusions followed by binding kinetics by cell binding. Mutations from the top performing variants in Fc fusion were combined to generate 6 humanized anti TCR VHH containing a range of 5 to 10 humanized residues. These were integrated into the final trispecific format. The final lead panel was assessed for cytotoxicity and T cell activation. All variants matched the criteria in final format. The following TCR V_HHs were selected: hu2.1 (SEQ ID NO: 43), hu2.2 (SEQ ID NO: 44), hu2.3 (SEQ ID NO: 45), hu2.4 (SEQ ID NO: 46), hu2.5 (SEQ ID NO: 47) and hu2.6 (SEQ ID NO: 48).

Example 8: Efficacy of LM1486 (CD20 TITAN) in Inflammatory Disease and Autoimmune Disorder Models

Methods

Human Peripheral Blood Mononuclear Cells (PBMCs)

Primary human PBMCs were isolated from the whole blood obtained from SLE or myositis patients. PBMCs were counted and 100K/well PBMCs were treated with varying concentrations of the CD20 TITAN molecule in a 96-well plate. PBMCs were cultured for three days at 37° C. in a CO₂ incubator under sterile conditions. The depletion of B cells was measured at the end of three days by flow cytometry analysis and IC₅₀ values were calculated using Prism GraphPad software.

Humanized Mouse Model

FIG. 29 depicts the method steps. NOD-scid IL2Rgamma^null (NSG) mice were obtained from Jackson lab at about 22 weeks post-engraftment, randomized and treated with either vehicle (PBS), NIP228 control at 1 mg/kg or either 0.03 mg/kg, 0.1 mg/kg, 0.3 mg/kg, or 1 mg/kg of LM1486 via intraperitoneal (i.p.) injection. 72 hours post-treatment, blood and tissues were collected and processed for further analysis.

The spleens were meshed through cell strainers with a syringe plunger with 5 mL of neutralize buffer (RPMI1640 (GIBCO) containing 5% FBS (GIBCO) and 2 mM EDTA (INVITROGEN) and centrifuged at 460 g for 5 minutes at 4° C. Cell pellets were resuspended with 1 mL of ACK lysis buffer (GIBCO) for 1 minute, then 2 mL of neutralize buffer was added before centrifugation at 460 g for 5 minutes at 4° C., the pellets were then resuspended with 2 mL of neutralize buffer.

The thymuses were meshed through cell strainers with a syringe plunger with 5 mL of neutralize buffer and centrifuged at 460 g for 5 minutes at 4° C. Cell Pellets were resuspended in 2 mL of neutralize buffer.

Bone marrow cells were flushed out from the femur and tibia from a hind leg using a 25G needle and passed through a cell strainer to remove any bone spicules, muscle or cell clumps, and red cells were lysed with ACK lysis buffer (GIBCO); after centrifugation the pellet was resuspended in cold neutralize buffer.

Blood samples were collected in EDTA pre-treated tube (BD) and centrifuged at 500 g for 10 minutes at 4° C. to separate the plasma from cells. The cell pellets were resuspended in 5 mL of ACK lysis buffer (GIBCO) for 90 seconds at room temperature to lyse the red cells. Washed with 5 mL of neutralize buffer and resuspended in 1 mL cold neutralize buffer.

Capsules and extra connected tissues were removed from all kidney samples by forceps and were gently crushed with a syringe plunger. Dissociation was obtained by using Miltenyi gentleMACS™ dissociation protocol.

In summary, samples were collected into a Miltenyi gentleMACS C tube with 2 mL Multi tissue dissociation medium and incubated on program “37 Multi E” for 30 minutes at 37° C. The cell suspensions were neutralized by adding 2 mL of neutralize buffer and filtered through 40 um cell strainers. After centrifugation, the pellets were resuspended in 40% percoll (GE HEALTHCARE) and centrifugated at 580 g for 20 minutes at room temperature. The cells on bottom were collected and washed with neutralize buffer, then resuspended into 1 mL of cold neutralize buffer. For flow cytometry analysis: 2 million cells were incubated with Live/Dead NIR 780 (THERMOFISHER) at 1:2000 dilution in 200 μl of RPMI1640 containing 2 g Fc blocker (BIOLEGEND) for 5 minutes at 4° C. Cells were washed with FACS buffer by centrifugation at 460 g for 5 minutes at 4° C. Pellets were resuspended in 50 μl of antibody mix for staining with anti-human CD3 BUV 395 (BD, 1:100dilution), anti-human CD19 BUV737 (BD, 1:200 dilution), anti-human CD8a BV711 (BIOLEGEND, 1:400 dilution), anti-human CD2 BB515 (BD, 1:1200 dilution), anti-human CD25 PE (BIOLEGEND, 1:50 dilution), anti-human CD4 PE-Cy7 (BIOLEGEND, 1:400 dilution), anti-human CD45 APC (BIOLEGEND, 1:200), anti-mouse CD45 BV510 (BIOLEGEND, 1:400 dilution) containing 2.5 μl Superbright staining buffer and incubating for 30 minutes on ice. Cells were washed twice with FACS buffer, and then fixed by adding 100 μl of 1× fixation buffer (EBIOSCIENCE) at room temperature for 20 minutes. After centrifugation, cells were resuspended in 200 μl FACS buffer. Flow cytometry was performed using Symphony A3 (BD BIOSCIENCE). Analysis was conducted with FLOJO software.

Lymph nodes were fixed for 48 hours in 4% PFA, dehydrated and embedded into paraffin blocks. Four micrometer thick tissue sections were cut and stained using an immunofluorescence panel with OPAL dyes (AKOYA BIOSCIENCES) for detection: anti-CD19 (EPR5906, Abcam; OPAL 520), anti-CD25 (EPR6452, Abcam; OPAL 570), anti-CD8 (D8A8Y, Cell Signaling Technology; OPAL 620), and anti-CD4 (EPR6855, Abcam; OPAL 480). Digital whole-slide images were generated using PhenoImager HT (AKOYA BIOSCIENCES) and cell classification and tissue area quantification was performed using VISIOPHARM v.2023.09.5.15777.

Results

Calculation of IC₅₀ values for B cell depletion and maximum depletion of B cells in SLE and myositis patients derived PBMCs are depicted in Table 6.

TABLE 6
B cell depletion and max depletion of B cells in SLE and Myositis
		B cell depletion IC50	% avg. Max B cell
	Patient ID	(pM)	depletion
	Myo 109785	107.5	96
	Myo 107370	2.8	94
	Myo 120323	7.1	62
	Myo 109852	1.1	86
	Myo 112243	3.8	89
	Myo 123136	13.3	68
	Myo 138502	38.1	84
	SLE 29144	8.5	81
	SLE 89313	1.2	88
	SLE 20643	11.5	60
	SLE 23199	1.4	27
	SLE 91744	19.1	41
	SLE 23150	0.5	63
	SLE 29942	0.3	41
	SLE 29941	1.8	75

FIGS. 27 and 28 demonstrate the efficacy and safety of LM1486 (CD20 TITAN). LM1486 showed superior and potent B cell depletion (FIGS. 27A and B) in cells from patients with SLE and myositis, while also demonstrating excellent safety with selective CD8+ T-cell activation (FIGS. 27E and F) and CD4 T-cell sparing activity (FIGS. 27C and D). FIGS. 28A and B demonstrate reduced cytokine release of IL-17A (FIG. 28A) and TNFα (FIG. 28B) in an assay of non-specific T-cell activation.

LM1486 (CD20 TITAN) achieved deep depletion of B cells in blood and tissue compartments in a humanized mouse model, 72 hours after 1 dose treatment in human-CD34+ cells engrafted NSG mice by flow cytometry. FIGS. 30A-F depict depletion of B cells in spleen (FIG. 30A), blood (FIG. 30B), bone marrow (FIG. 30C), kidney (FIG. 30D), thymus (FIG. 30E), and lymph nodes (FIG. 30F) in a humanized mouse model after a single dose LM1486 (CD20 TITAN). Depletion was more pronounced in peripheral blood and spleen.

Example 9: Efficacy of LM1486 (CD20 TITAN) in Cynomolgous Monkey Model

Method

Cynomolgus monkeys were dosed twice weekly with either vehicle (n=10) or CD20 TITAN at 3 mg/kg (n=10). The CD20 TITAN group were dosed for a total of 5 doses and necropsy performed on Day 16 (FIG. 44). Mean±SEM B-cell counts (CD19+) were evaluated by flow cytometry and presented for peripheral blood (time course from Day −14 to Day 16) and tissues (Day 16). Absolute circulating CD8+ and CD4+ T cells post first dose and activated CD8/CD4 (CD25+) fold difference over baseline were assessed during the first week of treatment. Finally, group mean plasma cytokine profiles (IL-6, TNFα and IFNγ) were also assessed as a time course from Day −14 to Day 16.

Results

In a cynomolgus monkey model, LM1486 (CD20 TITAN) exhibits potent B cell depletion activity in cynomolgus monkey with low systemic cytokine release and associated toxicity at 3 mg/kg IV. FIG. 45 shows the depletion of circulating CD19+ B cells (FIG. 45A) and tissue specific CD19+ B cells in spleen, bone marrow and lymph nodes (FIG. 45B). FIG. 46 shows shows T cell margination and preferential CD8 activation. Absolute circulating CD8+(FIG. 46A) and CD4+ T (FIG. 46B) cells post first dose and activated CD8/CD4 (CD25+) (FIG. 46C) show a fold difference over baseline during the first week of treatment. FIG. 47 shows the group mean plasma cytokine profiles for IL-6 (FIG. 47A), TNFα (FIG. 47B), and IFNγ (FIG. 47C) as a time course from Day −14 to Day 16.

Example 10: In Vitro and In Vivo Testing of LM1953 (STEAP2 TITAN)

The activity of the LM1953 (STEAP2, see FIG. 43D) binding protein disclosed herein was demonstrated.

In Vitro Cytolysis, T Cell Activation and Cytokine Release Profiles

Method

An assay was conducted to measure cytotoxic activity of LM1953. The assay was run for 3 days with the STEAP2+ prostate cancer lines 22Rv1 and C4-2 and PBMCs from 8 donors at an E:T ratio of 10:1 in an XCELLIGENCE assay. At the end of the cytolysis assay, T cell activation status was evaluated by flow cytometry and the cytokine release profiles were measured in the supernatant using a multiplexing assay.

Results

FIGS. 31A and B show potent in vitro cytolysis by LM1953. FIG. 31A shows the in vitro cytolytic activity of LM1953 on C4-2 cells and the corresponding EC₅₀. FIG. 31B shows the in vitro cytolytic activity of LM1953 on 22Rv1 cells and the corresponding EC₅₀. A CD8-biased T cell activation profile was induced by LM1953. FIG. 32A shows corresponding CD4 and CD8 T cell activation profiles as % of cells expressing CD25 on the cell surface in the C4-2 cytolysis assay and FIG. 32B in the 22Rv1 cytolysis assay. FIG. 33A-D show in vitro cytokine release profiles of LM1953 on C4-2 cells for IL-6 (FIG. 33A) and TNF-α (FIG. 33B), and on 22Rv1 cells for IL-6 (FIG. 33C) and TNF-α (FIG. 33D).

Anti-Tumor Efficacy of LM1953 in a Humanized Prostate Cancer Xenograft Cell Line Model

Method

Male NSG MHC I/II DKO mice were humanized i.v. with 10e6 PBMCs from 3 different donors (n=4/group) on day −7 then 5e6 22Rv1 tumor cells were injected s.c. on the flank on day 0. On days 2, 6, 10, 14, 17 and 21, 0.41 mg/kg of LM1953 was injected i.p. and tumor size was evaluated using calipers.

Results

FIG. 34 shows that LM1953 confers anti-tumor efficacy in vivo against STEAP2+ prostate cancer xenograft model 22Rv1.

Example 11: LRC150016 (LRRC15) Fab Variants with Improved Developability Profiles

Sequence variants of the LRC150016 V_H/V_L were designed to improve the developability and humanisation of the LRRC15 Fab. These variants were generated as human IgG1 format antibodies by standard methods and tested for retention of binding to human, mouse and cyno LRRC15 recombinant protein. Notably, it was found that variants with more human germline residues within H-CDR2 (SEQ ID NOs: 136, 147, 152 and 157) retained binding to LRRC15.

Selected LRRC15-binding Fab variants (see Table 7 below) were then formatted as T-cell engagers in the CD8/TCR format (see FIG. 15), with 2×LRRC15-binding Fab arms, 1×TCR binding VHH and 1×CD8 binding VHH. The CD8-binding sequence was replaced with a variant modified to remove two sequence liabilities identified in the original variant. One of these changes is in CDR1 by IGMT definition. These T-cell engagers were tested for their ability to induce CD8-mediated cytolysis of LRRC15-expressing tumour cell lines in co-culture with human PBMCs.

TABLE 7
	LRRC15 Fab	LRRC15 Fab
Candidate #	VH	VL	TCR VHH	CD8 VHH
LRC150016	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:
	31	32	42	52
TPP-46956	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:
	141	142	42	168
TPP-46957	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:
	148	142	42	168
TPP-46958	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:
	153	142	42	168
TPP-46959	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:
	159	160	42	168
TPP-46960	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:
	159	166	42	168
TPP-47826	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:
	153	142	172	168
TPP-49058	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:	SEQ ID NO:
	159	160	172	168

The affinities of the T-cell engagers to human, mouse and cyno LRRC15 recombinant proteins were measured by Surface Plasmon Resonance using a Single-Cycle Kinetics (SCK) method on a Biacore 8K+ system (CYTIVA).

The T-cell engagers were also tested for unwanted binding to cells transiently transfected with human EphB6. TPP-46959 was identified as having reduced binding to EphB6-expressing cells. This molecule differs from TPP-46960 at the L-CDR2 (SEQ ID NO: 158) where residue 55 is E55 in TPP-46959, compared to H55 in huM25 and N55 in TPP-46960.

LRRC15 (huM25) Fab Sequence Variants Affinity Determination—Single Cycle Kinetics Measurements.

The T-cell engagers were captured on a PROTEIN G chip and exposed to a series of increasing concentrations of human, mouse or cyno LRRC15 recombinant extracellular domain proteins (Acro Biosystems PROTEIN G). The fully corrected binding data were globally fitted to a 1:1 binding model to determine the K_D using the Biacore™ Insight Evaluation Software (CYTIVA). Coloured sensorgram trace represents the raw data and black line the global 1:1 fit to that data. Values for kinetics parameters is shown in Table 8 are averages for 3 or 4 replicate sets of measurements.

TABLE 8
T-cell	LRRC15	ka (M−1 s−1)	kd (s−1)	KD (nM)
engager	Species	Average	SD	Average	SD	Average	SD
TPP-46956	human	3.05E+05	2.61E+03	6.84E−04	1.10E−05	2.24	0.04
	cyno	1.70E+05	1.24E+03	8.18E−04	2.39E−05	4.80	0.1
	mouse	2.05E+05	5.16E+02	2.06E−04	7.55E−06	1.01	0.04
TPP-46957	human	2.64E+05	4.73E+03	6.00E−04	5.56E−06	2.27	0.06
	cyno	1.65E+05	3.23E+03	7.50E−04	4.02E−06	4.55	0.1
	mouse	1.98E+05	8.16E+02	1.88E−04	9.95E−06	0.953	0.05
TPP-46958	human	2.83E+05	4.25E+03	8.77E−04	3.54E−05	3.10	0.08
	cyno	1.70E+05	2.45E+03	1.09E−03	7.34E−05	6.40	0.5
	mouse	1.96E+05	4.04E+03	3.43E−04	5.59E−05	1.75	0.3
TPP-46959	human	3.51E+05	1.99E+04	4.68E−04	5.14E−05	1.34	0.2
	cyno	1.89E+05	2.40E+03	5.68E−04	6.13E−05	3.00	0.3
	mouse	2.38E+05	3.70E+03	1.44E−04	6.40E−06	0.608	0.04
TPP-46960	human	2.97E+05	1.04E+04	2.91E−04	1.65E−05	0.980	0.07
	cyno	1.70E+05	5.75E+03	3.79E−04	2.94E−05	2.24	0.2
	mouse	2.00E+05	3.32E+03	5.64E−05	2.56E−05	0.281	0.13

LRRC15 (huM25) Fab Sequence Variants Functional Assessment.

Whole PBMCs (3 donors) were cultured with LRRC15^pos Saos-2 Wt target cell line for 3 days at an E:T ratio of 10 to 1. LRC150016 and its LRRC15 Fab sequence variants mediated T cell activation, as measured by % CD25 surface expression. Tabular data (Table 9 below) refers TCE molecule EC₅₀ (pM) and E_max (0%) values for CD8 T-cells activation.

	TABLE 9
	TCE	EC50 (pM)	Emax (%)
	LRC150016	3.3	52.31
	TPP-46956	3.9	53.32
	TPP-46957	3.7	49.69
	TPP-46958	3.2	55.17
	TPP-46859	2.0	54.63
	TPP-46960	2.8	55.06

Results are shown in FIG. 35A, values are mean (±SEM) frequency of T-cell activation.

Dose-response curves demonstrated that both LRC150016 and the LRRC15 Fab sequence variant TCEs mediate T-cell activation in a dose dependent manner, as measured by the CD25 surface expression, following 3 days of whole PBMCs co-culture with LRRC15^pos Saos-2 Wt Target cells. Similar to LRC150016, all five LRRC15 Fab sequence variant TCEs selectively activate CD8+ T-cells, while showing no activation of CD4+ T cells. Values shown here are the mean (±SEM) frequency of CD25⁺ cells among CD8 and CD4 T-cells. n=3 donors from a representative experiment.

In-Vitro Cytolytic Activity of LRC150016 and its LRRC15 Fab Sequence Variants Against LRRC15^pos Saos-2 WT Target Cell Line

Target cytolysis was assessed using CELL-TITER-GLO. Tabular data refers to TCE molecule EC₅₀ (pM) and E_max (%) values for target cell cytolysis, see Table 10.

Results are shown in FIG. 35B. Dose-response curves show that compared to LRC150016, all five LRRC15 Fab sequence variant TCEs mediate similar or slightly higher levels of target (Saos-2 Wt) cell cytolysis, as measured by the cytolysis E_max values, after 3 days of Saos-2 target cell co-culture with whole PBMC effector cells. Target cell cytolysis was assessed using CELL-TITER-GLO. Nip 228 control TCE did not mediate any target cell cytotoxicity. Values shown here are the mean (±SEM) frequency of target cell cytolysis (cytotoxicity). n=3 donors from a representative experiment.

	TABLE 10
	TCE	EC50 (pM)	Emax (%)
	LRC150016	6.7	46.84
	TPP-46956	8.7	59.65
	TPP-46957	5.3	49.82
	TPP-46958	6.1	57.72
	TPP-46859	4.4	60.02
	TPP-46960	4.3	60.57

Evaluation of Cytokine (IL-6) Profile in the Supernatant of Whole PBMCs Co-Culture Assay.

Whole PBMCs were cultured with LRRC15^pos Saos-2 Wt target cells either in the presence of LRC150016 or one of five LRRC15 fab sequence variant TCEs for a period of 3 days at an ET ratio of 10 to 1. IL-6, a surrogate for cytokine release syndrome (CRS), was measured in the culture supernatant after 3-days of co-culture using standard ELISA. Values shown here are the mean (±SEM) frequency of IL-6 (pg/mL) in the cell culture supernatant. Results are shown in FIG. 35C, the LRRC15 variants had a similar effect on IL-6 levels as LRC150016.

EphB6 Off-Target Binding Assessment

Ad293 cells were transiently transfected with a plasmid expressing human EphB6. Binding of the LRRC15 T-cell engagers was evaluated by flow cytometry.

Results are shown in FIG. 36, TPP-46959 showed reduced off-target binding to EphB6 expressing AD293 cells, compared to other sequence variants.

Example 12: TPP-42197 (BCMA TITAN) Depletion of the Plasmablast/Plasma Cells in Primary Assays

Method

Healthy donor PBMCs were thawed, counted and naïve B cells were separated by magnetic beads. The cells were cultured with 1 g/ml CpG and 100U/ml IL-2 for 5 days in RPMI media enriched with ATCC to induce B cell differentiation into plasmablasts. Following 5 days of differentiation, autologous PBMCs were thawed and co-cultured with differentiated plasmablasts at a ratio of 1:5 target to effector cells. To maintain this ration, the % of CD3+ T cells per donor was assessed by flow cytometry and the number of PBMCs to be cultured, was determined so it will keep this ratio between plasmablasts and T cells (50,000 plasmablasts with 250,000 CD3+ T cells). The cells were stimulated with increasing concentrations (0.0000003 nM to 20 nM) of either TPP-42197 or the comparator, “Comparator T” (CD3×BCMA TCE) for 48h. At the end of the experiment, the supernatant was collected and the levels of cytokine secretion from the cells was assessed by MSD ELISA. The level of plasmablast cytolysis as well as the level of T cell activation for both CD8+ cytotoxic T cells and CD4+ helper T cells were measured by flow cytometry.

Results

TPP-42197 was found to have a better potency than Comparator T and a better EC₅₀ (2.2 pM versus 415 pM respectively), see FIG. 38. In addition, TPP-42197 induced a selective activation of CD8+ T cells and no activation of CD4+ T cells, the latter being major contributors of cytokine secretion (FIG. 39A-B). In contrast, Comparator T induced activation of both CD8+ and CD4+ T cells. The cytokine profile of TPP-42197 was shown to be superior to that of Comparator T with lower cytokine secretion in general and particularly in those involved in cytokine release syndrome (CRS) events (see FIGS. 40A-F)

TCP-42197Fully Depletes Engrafted Human BCMA+ Cells in Bone Marrow and Spleen

To assess the ability of TCP-42197 to deplete primary B cells expressing BCMA in vivo, a pharmacodynamic (PD) study was performed in the xenogeneic model of graft versus host disease (XenoGvHD). To prepare for this XenoGvHD PD study, healthy human donor peripheral blood mononuclear cells (PBMCs) were isolated from fresh Leukopak (StemExpress™) and stored frozen (liquid nitrogen tank) until the engraftment.

On Day −1, 8-10 week old female NOD scid gamma mice (NSG; Jackson Laboratories) were pre-conditioned with sub-lethal whole-body irradiation (1 Gy). On Day 0, 15 million total PBMC from a healthy human donor was injected intravenously. On day 9, mice received either phosphate buffered saline (PBS), 1 mg/kg TCP-42197, 10 mg/kg TCP-42197 or 10 mg/kg isotype control, intraperitoneally (n=5-9 mice/group). On Day 12, mice were euthanized, and tissues were collected for FACS analysis to determine whether these TCEs depleted the BCMA+ cells present in this in vivo model.

Plasmablasts, defined by flow cytometric analysis as human CD45⁺CD27⁺CD38⁺CD3⁻CD56⁻CD14⁻CD16⁻, were depleted in mice three days following a single treatment of TCP-42197 in both the bone marrow and in the spleen (FIGS. 37A and 37B). Serum human IgG was significantly lower in the TCP-42197 treatment groups compared to the isotype control treated group (FIG. 37C), reflecting the absence of BCMA+ antibody secreting cells in the TCP-42197 treated mice.

Claims

1. A binding protein comprising four polypeptide chains that form two tumor-associated antigen (TAA) binding sites, a T cell receptor binding site, and a T cell co-stimulatory molecule binding site, wherein the first and second polypeptide chains have a structure represented by the formula: VL-CL a third polypeptide chain has a structure represented by the formula: VH1-CH1-VHHa-FCa and a fourth polypeptide chain has a structure represented by the formula: VH1-CH1-VHHb-FCb wherein the first polypeptide and the third polypeptide form the first of the two TAA binding sites, and the second polypeptide and the fourth polypeptide form the second of the two TAA binding sites, andwherein: VL is an immunoglobulin light chain variable domain and VH1 is an immunoglobulin heavy chain variable domain that together form a TAA binding domain that specifically binds a tumor-associated antigen;CL is an immunoglobulin light chain constant domain;CH1 is an immunoglobulin CH1 heavy chain constant domain;VHHa is a single chain variable domain that specifically binds a T cell receptor;VHHb is a single chain variable domain that specifically binds T cell co-stimulatory molecule;FCa is CH2a and CH3a immunoglobulin heavy chain constant domains; andFCb is CH2b and CH3b immunoglobulin heavy chain constant domains.

2. The binding protein of claim 1, further comprising L1, a linker positioned between CH1 and VHHa on the third polypeptide chain, and L2, a linker positioned between VHHa and the FCa on the third polypeptide chain, wherein L1 and L2 are each independently a linker or are absent.

3. The binding protein of either claim 1 or claim 2, further comprising L3, a linker positioned between CH1 and VHHb on the fourth polypeptide chain, and L4, a linker positioned between VHHb and FCb on the fourth polypeptide chain, wherein L3 and L4 are each independently a linker or are absent.

4. The binding protein of any one of claims 1-3 further comprising H1, an immunoglobulin hinge region positioned between CH1 and VHHa on the third polypeptide chain, and H2, an immunoglobulin hinge region positioned between VHHa and the FCa on the third polypeptide chain, wherein H1 and H2 are each independently an immunoglobulin hinge region or are absent.

5. The binding protein of claim 4, wherein H1 comprises SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 90 or is absent, and wherein H2comprises SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 90 or is absent.

6. The binding protein of any one of claims 1-5, further comprising H3, an immunoglobulin hinge region positioned between CH1 and VHHb on the fourth polypeptide chain, and H4, an immunoglobulin hinge region positioned between VHHb and the FCb on the fourth polypeptide chain, wherein H3 and H4 are each independently an immunoglobulin hinge region or are absent.

7. The binding protein of claim 6, wherein H3 comprises SEQ ID NO: 53, DK (SEQ ID NO: 54), SEQ ID NO: 90 or is absent, and wherein H4 comprises SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 91 or is absent.

8. The binding protein of any one of claims 1-7, wherein the first and second polypeptide chains have a structure represented by the formula: VL-CL a third polypeptide chain has a structure represented by the formula: VH1-CH1-H1-L1-VHHa-H2-L2-FCa orVH1-CH1-H1-L1-VHHa L2-H2-FCa and a fourth polypeptide chain has a structure represented by the formula: VH1-CH1-H3-L3-VHHb-H4-L4-FCb orVH1-CH1-H3-L3-VHHb-L4-H4-FCb.

9. The binding protein of any one of claims 1-8, wherein FCa and/or FCb is from an IgG antibody.

10. The binding protein of claim 9, wherein the FCa and/or the Fe is from an IgG1 antibody.

11. The binding protein of any one of claims 1-10, wherein the binding protein activates T cells only when bound to a tumor associated antigen at one or both of the tumor-associated antigen binding sites.

12. The binding protein of any one of claims 2-11, wherein L1, L2, L3, and/or L4 comprise one or more repeats of the amino acid sequence of SEQ ID NO: 37 and/or SEQ ID NO: 38.

13. The binding protein of any one of claims 1-12, wherein the VHHa comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2) and a heavy chain CDR3 (HCDR3), comprising the amino acid sequences of SEQ ID NO: 39, SEQ ID NO: 40 and SEQ ID NO: 41, respectively.

14. The binding protein of any one of claims 1-12, wherein the VHHa comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2) and a heavy chain CDR3 (HCDR3) comprising the amino acid sequence of SEQ ID NO: 62, SEQ ID NO: 40 and SEQ ID NO: 41, respectively.

15. The binding protein of any one of claims 1-14, wherein the VHHa Comprises the amino acid sequence at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 42-48, or any one of SEQ ID NOs: 42-48.

16. The binding protein of any one of claims 1-12, wherein the VHHa comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2) and a heavy chain CDR3 (HCDR3), comprising the amino acid sequences of SEQ ID NO: 173, SEQ ID NO: 174, and SEQ ID NO: 175 respectively.

17. The binding protein of any one of claims 1-12 or 16, wherein the VHHa Comprises the amino acid sequence at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 172, or SEQ ID NO: 172.

18. The binding protein of any one of claims 1-17, wherein the T cell costimulatory molecule is CD8.

19. The binding protein of any one of claims 1-18, wherein the VHHb comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2) and a heavy chain CDR3 (HCDR3) comprising the amino acid sequences of SEQ ID NO: 49, SEQ ID NO: 50 and SEQ ID NO: 51, respectively.

20. The binding protein of any one of claims 1-19, wherein the VHHb Comprises an amino acid sequence at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 52, or SEQ ID NO: 52.

21. The binding protein of any one of claims 1-18, wherein the VHHb comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2) and a heavy chain CDR3 (HCDR3) comprising the amino acid sequences of SEQ ID NO: 169, SEQ ID NO: 170 and SEQ ID NO: 171, respectively.

22. The binding protein of any one of claims 1-21, wherein the VHHb Comprises an amino acid sequence at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 168, or SEQ ID NO: 168.

23. The binding protein of any one of claims 1-22, wherein CH3a and CH3b in each of FCa and FCb comprise modifications to facilitate heterodimerization of FCa and FCb.

24. The binding protein of claim 23, wherein the modifications are substitutions to generate a knob in one of FCa and FCb and a hole in the other of FCa and FCb, wherein the knob is a substitution to tryptophan at position 366, and wherein the hole is a substitution of one or more of the following: (i) a substitution to valine at position 407;(ii) a substitution to serine at position 366; and(iii) a substitution to alanine at position 368and wherein the numbering is according to the Eu index.

25. The binding protein of claim 24, wherein the FCa or the FCb containing the knob further comprises a cysteine at position 354 and/or the FCa or the FCb containing the hole comprises a cysteine at position 349, and wherein the numbering is according to the Eu index.

26. The binding protein of any one of claims 1-25 wherein the CH2a and CH2b immunoglobulin heavy chain constant domains each comprise the following substitutions: E233P/L234V/L235A/G236del/S267K, and wherein the numbering is according to the Eu index.

27. The binding protein of any one of claims 1-26, wherein either the CH3a immunoglobulin heavy chain constant domain or CH3b immunoglobulin heavy chain constant domain comprises a H435R and Y436F substitution, and wherein the numbering is according to the Eu index.

28. The binding protein of any one of claims 1-27, wherein FCa comprises the amino acid sequence of SEQ ID NO: 56 or SEQ ID NO: 57.

29. The binding protein of any one of claims 1-28, wherein FCb comprises the amino acid sequence of SEQ ID NO: 58.

30. The binding protein of any one of claims 1-29, wherein the third polypeptide chain comprises either SEQ ID NO: 59 or SEQ ID NO: 60 and the fourth polypeptide chain comprises SEQ ID NO: 61.

31. The binding protein of any one of claims 1-30, wherein the CL of the first polypeptide chain and/or the CL of the second polypeptide chain comprises a constant light chain lambda region (CLλ); and the binding protein comprises at least one lambda charge pair wherein the lambda charge pair is either between the CLλ of the first polypeptide chain and a corresponding CH1 that is the CH1 of the third polypeptide chain, or the lambda charge pair is between the CLλ of the second polypeptide and a corresponding CH1 that is the CH1 of the fourth polypeptide chain, and where the lambda charge pair is located at one or more of the following pairs of positions: (i) position 117 in the CLλ and position 141 in the corresponding CH1;(ii) position 117 in the CLλ and position 185 in the corresponding CH1;(iii) position 119 in the CLλ and position 128 in the corresponding CH1;(iv) position 134 in the CLλ and position 128 in the corresponding CH1;(v) position 134 in the CLλ and position 145 in the corresponding CH1;(vi) position 134 in the CLλ and position 183 in the corresponding CH1;(vii) position 136 in the CLλ and position 185 in the corresponding CH1;(viii) position 178 in the CLλ and position 173 in the corresponding CH1; and(ix) position 117 in the CLλ and position 187 in the corresponding CH1;wherein the lambda charge pair comprises a positively charged amino acid residue selected from arginine, lysine and histidine located at one of the positions in the lambda charge pair and a negatively charged amino acid residue selected from aspartic acid, glutamic acid, serine and threonine located at the other position in the lambda charge pair; andwherein the numbering is according to the Eu index.

32. The binding protein of claim 31, wherein the lambda charge pair is selected from the following list: (a) arginine at position 117 of the CLλ and aspartic acid at position 141 in the corresponding CH1;(b) arginine at position 117 of the CLλ and glutamic acid at position 141 in the corresponding CH1;(c) arginine at position 117 of the CLλ and serine at position 141 in the corresponding CH1;(d) arginine at position 117 of the CLλ and threonine at position 141 in the corresponding CH1;(e) lysine at position 117 of the CLλ and aspartic acid at position 141 in the corresponding CH1;(f) lysine at position 117 of the CLλ and glutamic acid at position 141 in the corresponding CH1;(g) lysine at position 117 of the CLλ and serine at position 141 in the corresponding CH1; and(h) lysine at position 117 of the CLλ and threonine at position 141 in the corresponding CH1.

33. The binding protein of either claim 31 or claim 32, wherein the CLλ of the first polypeptide chain and/or CLλ of the second polypeptide chain each comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NOs: 98-99.

34. The binding protein of any one of claims 1-33, wherein the CL of the first polypeptide chain and/or the CL of the second polypeptide chain comprises a constant light chain kappa region (CLκ); and a kappa charge pair wherein the kappa charge pair is either between the CLκ of the first polypeptide chain and a corresponding CH1 that is the CH1 of the third polypeptide chain, or the kappa charge pair is between the CLκ of the second polypeptide and a corresponding CH1 that is the CH1 of the fourth polypeptide chain; wherein the kappa charge pair is located at position 117 in the CLκ and position 141 in the corresponding CH1 and comprises a positively charged amino acid residue selected from arginine, lysine, and histidine located at one of the positions in the kappa charge pair, and a negatively charged amino acid residue selected from aspartic acid, glutamic acid, serine and threonine located at the other position in the kappa charge pair.

35. The binding protein of claim 34, wherein the negatively charged amino acid residue in the kappa charge pair is located at position 133 of the CLκ, and the positively charged amino acid residue is located at position 183 of the corresponding CH1, optionally wherein the negatively charged amino acid residue at position 133 of the CLκ is a glutamic acid, and optionally wherein the positively charged amino acid residue at position 183 of the corresponding CH1 is a lysine.

36. The binding protein of either claim 34 or claim 35, wherein the CLκ of the first polypeptide chain or the second polypeptide chain comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 100.

37. The binding protein of any one of claims 34-36 wherein the corresponding CH1 in each of the one or more lambda charge pairs comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to SEQ ID NO: 95-97, and/or the corresponding CH1 in the kappa charge pair comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to SEQ ID NO: 100.

38. The binding protein of any one of claims 31-37, wherein the CLλ comprises an amino acid sequence according to SEQ ID NO: 105, and the corresponding CH1 Comprises an amino acid sequence according to SEQ ID NO: 104 or SEQ ID NO: 220.

39. The binding protein of any one of claims 1-38, wherein either: (i) a disulfide link between the CL of the first polypeptide chain and the CH1 of the third polypeptide chain is formed between a pair of cysteines engineered into the CL of the first polypeptide chain and the CH1 of the third polypeptide chain, and a disulfide link between the CL of the second polypeptide chain and the CH1 of the fourth polypeptide chain is formed between a pair of native cysteines; or(ii) a disulfide link between the CL of the second polypeptide chain and the CH1 of the fourth polypeptide chain is formed between a pair of cysteines engineered into the CL of the second polypeptide chain and the CH1 of the fourth polypeptide chain, and a disulfide link between the CL of the first polypeptide chain and the CH1 of the third polypeptide chain is formed between a pair of native cysteines.

40. The binding protein of claim 39, wherein the pair of cysteines are engineered into either: (i) the CL of the first polypeptide chain and the CH1 of the third polypeptide chain, wherein the CL of the first polypeptide chain is a CLλ and wherein the pair of engineered cysteines are located at position 122 of the CLλ of the first polypeptide chain and position 126 of the CH1 of the third polypeptide chain, and wherein the CL, of the first polypeptide chain comprises a non-cysteine residue at position 212 and the CH1 of the third polypeptide chain comprises a non-cysteine residue at position 220, optionally wherein the non-cysteine residues are valines; or(ii) the CL of the second polypeptide chain and the CH1 of the fourth polypeptide chain, wherein the CL of the first polypeptide chain is a CLλ and wherein the pair of engineered cysteines are located at position 122 of the CLλ of the second polypeptide chain and position 126 of the CH1 of the fourth polypeptide chain, and wherein the CLλ of the second polypeptide chain comprises a non-cysteine residue at position 212 and the CH1 of the fourth polypeptide chain comprises a non-cysteine residue at position 220, optionally wherein the non-cysteine residues are valines.

41. The binding protein of claim 39, wherein the pair of cysteines are engineered into either: (i) the CL of the first polypeptide chain and the CH1 of the third polypeptide chain, wherein the CL of the first polypeptide chain is a CLκ and wherein the pair of engineered cysteines are located at position 121 of the CLκ of the first polypeptide chain and position 126 of the CH1 of the third polypeptide chain, and wherein the CLκ of the first polypeptide chain comprises a non-cysteine residue at position 214 and the CH1 of the third polypeptide chain comprises a non-cysteine residue at position 220, optionally wherein the non-cysteine residues are valines; or(ii) the CL of the second polypeptide chain and the CH1 of the fourth polypeptide chain, wherein the CL of the first polypeptide chain is a CLκ and wherein the pair of engineered cysteines are located at position 121 of the CLκ of the second polypeptide chain and position 126 of the CH1 of the fourth polypeptide chain, and wherein the CLκ of the second polypeptide chain comprises a non-cysteine residue at position 214 and the CH1 of the fourth polypeptide chain comprises a non-cysteine residue at position 220, optionally wherein the non-cysteine residues are valines.

42. The binding protein of any one of claims 39-41, wherein the CLλ comprises an amino acid sequence according to SEQ ID NO: 107, and the corresponding CH1 comprises an amino acid sequence according to SEQ ID NO: 106.

43. The binding protein of any one of claims 1-42, wherein the tumor-associated antigen (TAA) is CD20, Glypican-3 (GPC3), or leucine rich repeat containing 15 (LRRC15).

44. The binding protein of any one of claims 1-42, wherein the tumor-associated antigen (TAA) is B-cell maturation antigen (BCMA) or six-transmembrane epithelial antigen of prostate-2 (STEAP2).

45. The binding protein of any one of claims 1-43, wherein the TAA binding domain binds to CD20 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a light chain CDR3 (LCDR3), comprising the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, respectively.

46. The binding protein of any one of claims 1-43, wherein the TAA binding domain binds to CD20 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a light chain CDR3 (LCDR3), comprising the amino acid sequences of SEQ ID NO: 182, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, respectively.

47. The binding protein of claim 45 or claim 46, wherein the TAA binding domain comprises a VH1 domain and a VL domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 7 and SEQ ID NO: 8, respectively; or comprises a VH chain according to SEQ ID NO: 7 and a VL according to SEQ ID NO: 8.

48. The binding protein of any one of claims 1-43 wherein the TAA binding domain binds to GPC3 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, and SEQ ID NO: 18, respectively.

49. The binding protein of claim 48, wherein the TAA binding domain comprises a VH1 domain and a VL domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 19 and SEQ ID NO: 20, respectively; or comprises a VH chain according to SEQ ID NO: 19 and a VL according to SEQ ID NO: 20.

50. The binding protein of any one of claims 1-43, wherein the TAA binding domain binds to LRRC15 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO: 30 respectively.

51. The binding protein of claim 50, wherein the TAA binding domain comprises a VH1 domain and a VL domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 31 and SEQ ID NO: 32, respectively; or comprises a VH chain according to SEQ ID NO: 31 and a VL according to SEQ ID NO: 32.

52. The binding protein of any one of claims 1-43, wherein the TAA binding domain binds to LRRC15 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, and SEQ ID NO: 140 respectively.

53. The binding protein of claim 52, wherein the TAA binding domain comprises a VH1 domain and a VL domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 141 and SEQ ID NO: 142, respectively; or comprises a VH chain according to SEQ ID NO: 141 and a VL according to SEQ ID NO: 142.

54. The binding protein of any one of claims 1-43, wherein the TAA binding domain binds to LRRC15 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 147, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, and SEQ ID NO: 140 respectively.

55. The binding protein of claim 54, wherein the TAA binding domain comprises a VH1 domain and a VL domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 148 and SEQ ID NO: 142, respectively; or comprises a VH chain according to SEQ ID NO: 148 and a VL according to SEQ ID NO: 142.

56. The binding protein of any one of claims 1-43, wherein the TAA binding domain binds to LRRC15 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 152, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, and SEQ ID NO: 140 respectively.

57. The binding protein of claim 56, wherein the TAA binding domain comprises a VH1 domain and a VL domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 153 and SEQ ID NO: 142, respectively; or comprises a VH chain according to SEQ ID NO: 153 and a VL according to SEQ ID NO: 142.

58. The binding protein of any one of claims 1-43, wherein the TAA binding domain binds to LRRC15 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 157, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 158, and SEQ ID NO: 140 respectively.

59. The binding protein of claim 58, wherein the TAA binding domain comprises a VH1 domain and a VL domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 159 and SEQ ID NO: 160, respectively; or comprises a VH chain according to SEQ ID NO: 159 and a VL according to SEQ ID NO: 160.

60. The binding protein of any one of claims 1-43, wherein the TAA binding domain binds to LRRC15 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 157, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 165, and SEQ ID NO: 140 respectively.

61. The binding protein of claim 60, wherein the TAA binding domain comprises a VH1 domain and a VL domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 159 and SEQ ID NO: 166, respectively; or comprises a VH chain according to SEQ ID NO: 159 and a VL according to SEQ ID NO: 166.

62. The binding protein of any one of claims 1-42 or 44, wherein the TAA binding domain binds to BCMA and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, and SEQ ID NO: 116, respectively.

63. The binding protein of claim 62, wherein the TAA binding domain comprises a VH1 domain and a VL domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 117 and SEQ ID NO: 118, respectively; or comprises a VH chain according to SEQ ID NO: 117 and a VL according to SEQ ID NO: 118.

64. The binding protein of any one of claims 1-42 or 44, wherein the TAA binding domain binds to STEAP2 and comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, and SEQ ID NO: 128, respectively.

65. The binding protein of claim 64, wherein the TAA binding domain comprises a VH1 domain and a VL domain that is at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 129 and SEQ ID NO: 130, respectively; or comprises a VH chain according to SEQ ID NO: 129 and a VL according to SEQ ID NO: 130.

66. The binding protein of any one of claims 1-43 or 45-65 comprising the amino acid of sequences of SEQ ID NO: 9, SEQ ID NO: 11, and SEQ ID NO: 12.

67. The binding protein of any one of claims 1-43 or 45-65 comprising the amino acid of sequences of SEQ ID NO: 21, SEQ ID NO: 23, and SEQ ID NO: 24.

68. The binding protein of any one of claims 1-43 or 45-65 comprising the amino acid of sequences of SEQ ID NO: 33, SEQ ID NO: 35, and SEQ ID NO: 36.

69. The binding protein of any one of claims 1-42 or 44-65 comprising the amino acid of sequences of SEQ ID NO: 119, SEQ ID NO: 121, and SEQ ID NO: 122.

70. The binding protein of any one of claims 1-42 or 44-65 comprising the amino acid of sequences of SEQ ID NO: 131, SEQ ID NO: 134, and SEQ ID NO: 133.

71. The binding protein of any one of claims 1-43 or 45-65 comprising the amino acid of sequences of SEQ ID NO: 143, SEQ ID NO: 145, and SEQ ID NO: 146.

72. The binding protein of any one of claims 1-43 or 45-65 comprising the amino acid of sequences of SEQ ID NO: 143, SEQ ID NO: 150, and SEQ ID NO: 151.

73. The binding protein of any one of claims 1-43 or 45-65 comprising the amino acid of sequences of SEQ ID NO: 143, SEQ ID NO: 155, and SEQ ID NO: 156.

74. The binding protein of any one of claims 1-43 or 45-65 comprising the amino acid of sequences of SEQ ID NO: 161, SEQ ID NO: 163, and SEQ ID NO: 164.

75. The binding protein of any one of claims 1-43 or 45-65 comprising the amino acid of sequences of SEQ ID NO: 167, SEQ ID NO: 163, and SEQ ID NO: 164.

76. The binding protein of any one of claims 1-43 or 45-65 comprising the amino acid of sequences of SEQ ID NO: 161, SEQ ID NO: 177, and SEQ ID NO: 164.

77. The binding protein of any one of claims 1-43 or 45-65 comprising the amino acid of sequences of SEQ ID NO: 143, SEQ ID NO: 176, and SEQ ID NO: 156.

78. The binding protein of claim 66, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 9, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 11, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 12.

79. The binding protein of claim 67, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 21, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 23, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 24.

80. The binding protein of claim 68, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 33, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 35, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 36.

81. The binding protein of claim 69, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 119, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 121, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 122.

82. The binding protein of claim 70, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 131, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 134, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 133.

83. The binding protein of claim 71, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 143, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 145, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 146.

84. The binding protein of claim 72, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 143, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 150, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 151.

85. The binding protein of claim 73, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 143, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 155, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 156.

86. The binding protein of claim 74, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 161, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 163, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 164.

87. The binding protein of claim 75, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 167, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 163, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 164.

88. The binding protein of claim 76, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 161, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 177, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 164.

89. The binding protein of claim 77, wherein the first and second polypeptide chains comprise the amino acid sequence of SEQ ID NO: 143, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 176, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 156.

90. A pharmaceutical composition comprising the binding protein of any one of claims 1-89 and a pharmaceutically acceptable carrier.

91. An isolated nucleic acid molecule encoding the binding protein of any one of claims 1-89.

92. The isolated nucleic acid molecule according to claim 91 comprising the nucleotide sequence according to any one of SEQ ID NOs: 184-186.

93. The isolated nucleic acid molecule according to claim 91 comprising the nucleotide sequence according to any one of SEQ ID NOs: 187-189.

94. The isolated nucleic acid molecule according to claim 91 comprising the nucleotide sequence according to any one of SEQ ID NOs: 190-192.

95. The isolated nucleic acid molecule according to claim 91 comprising the nucleotide sequence according to any one of SEQ ID NOs: 193-195.

96. The isolated nucleic acid molecule according to claim 91 comprising the nucleotide sequence according to any one of SEQ ID NOs: 196-198.

97. The isolated nucleic acid molecule according to claim 91 comprising the nucleotide sequence according to any one of SEQ ID NOs: 199-201.

98. The isolated nucleic acid molecule according to claim 91 comprising the nucleotide sequence according to any one of SEQ ID NOs: 202-204.

99. The isolated nucleic acid molecule according to claim 91 comprising the nucleotide sequence according to any one of SEQ ID NOs: 205-207.

100. The isolated nucleic acid molecule according to claim 91 comprising the nucleotide sequence according to any one of SEQ ID NOs: 208-210.

101. The isolated nucleic acid molecule according to claim 91 comprising the nucleotide sequence according to any one of SEQ ID NOs: 211-213.

102. The isolated nucleic acid molecule according to claim 91 comprising the nucleotide sequence according to any one of SEQ ID NOs: 214-216.

103. The isolated nucleic acid molecule according to claim 91 comprising the nucleotide sequence according to any one of SEQ ID NOs: 217-219.

104. A vector comprising the isolated nucleic acid molecule of any one of claims 91-103.

105. A method of treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of claims 1-89 or the pharmaceutical composition of claim 90 to the subject.

106. A method of treating an inflammatory disease and/or autoimmune disorder in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of claims 1-89 or the pharmaceutical composition of claim 90 to the subject.

107. The method of claim 105, wherein the cancer is a B-cell malignancy, liver cancer, hepatocellular carcinoma (HCC), lung cancer, non-small cell lung cancer (NSCLC), squamous non-small cell lung cancer (sqNSCLC), ovarian cancer, clear cell ovarian cancer, carcinoma, Merkel cell carcinoma, gastric cancer, hepatoblastoma, nephroblastoma, melanoma, sarcoma, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial cell carcinoma, osteosarcoma, glioblastoma, thyroid cancer, multiple myeloma, prostate cancer or Ewing's sarcoma.

108. The method of claim 106, wherein the inflammatory disease and/or autoimmune disorder is scleroderma, systemic lupus erythematosus (SLE), myositis, rheumatoid arthritis (RA), Sjogren's syndrome, or anti-neutrophil cytoplasmic autoantibody (ANCA) vasculitis.

109. The method of claim 105, wherein the tumor-associated antigen of the binding protein is CD20 and the cancer is a B-cell malignancy.

110. The method of claim 109, wherein the B-cell malignancy is non-Hodgkin's lymphoma (NHL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Burkett lymphoma, primary mediastinal large B cell lymphoma (PMBCL) or small lymphocytic lymphoma (SLL).

111. The method of claim 105, wherein the tumor-associated antigen of the binding protein is Glypican-3 (GPC3), and the cancer is liver cancer, hepatocellular carcinoma (HCC), non-small cell lung cancer (NSCLC), squamous non-small cell lung cancer (sqNSCLC), ovarian cancer, clear cell ovarian cancer, carcinoma, Merkel cell carcinoma, gastric cancer, hepatoblastoma, or nephroblastoma.

112. The method of claim 105, wherein the tumor-associated antigen of the binding protein is leucine rich repeat containing 15 (LRRC15), and the cancer is melanoma, sarcoma, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial cell carcinoma, osteosarcoma, glioblastoma, lung cancer, non-small cell lung cancer, or thyroid cancer.

113. The method of claim 105, wherein the tumor-associated antigen of the binding protein is B cell maturation antigen (BCMA), and the cancer is multiple myeloma.

114. The method of claim 106, wherein the tumor-associated antigen of the binding protein is B cell maturation antigen (BCMA), and the autoimmune disorder is scleroderma, systemic lupus erythematosus (SLE), myositis, rheumatoid arthritis (RA) or Sjogren's syndrome.

115. The method of claim 105, wherein the tumor-associated antigen of the binding protein is STEAP2, and the cancer is prostate cancer or Ewing's sarcoma.

116. A method of treating systemic lupus erythematosus (SLE) in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of claims 1-89 or the pharmaceutical composition of claim 90 to the subject.

117. A method of treating myositis in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of claims 1-89 or the pharmaceutical composition of claim 90 to the subject.

118. A method of treating scleroderma in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of claims 1-89 or the pharmaceutical composition of claim 90 to the subject.

119. A method of treating rheumatoid arthritis (RA) in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of claims 1-89 or the pharmaceutical composition of claim 90 to the subject.

120. A method of treating Sjogren's syndrome in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of claims 1-89 or the pharmaceutical composition of claim 90 to the subject.

121. A method of treating anti-neutrophil cytoplasmic autoantibody (ANCA) vasculitis in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of claims 1-89 or the pharmaceutical composition of claim 90 to the subject.

122. A method of treating a B cell malignancy in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of claims 1-89 or the pharmaceutical composition of claim 90 to the subject.

123. A method of treating hepatocellular carcinoma in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of claims 1-89 or the pharmaceutical composition of claim 90 to the subject.

124. A method of treating osteosarcoma in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of claims 1-89 or the pharmaceutical composition of claim 90 to the subject.

125. A method of treating multiple myeloma in a subject in need thereof, comprising administering a therapeutically effective amount of the binding protein of any one of claims 1-89 or the pharmaceutical composition of claim 90 to the subject.

126. The method of any one of claims 105 to 125, wherein the binding protein preferentially activates a subset of T cells in the subject.

127. The method of claim 126, wherein the subset of T cells are CD8+ T cells.

128. The method of claim 127, wherein the CD8+ T cells are preferentially activated as compared to CD4+ T cells.

129. The method of any one of claims 126-128, wherein the activation of T cells is determined by measuring the percentage of surface CD25+ T cells.

130. The method of claim 129, wherein the percentage of surface CD25+ T cells that are CD8+ T cells is higher than the percentage of surface CD25+ T cells that are CD4+ T cells.

131. The method of any one of claims 105-130, wherein the binding protein is administered by subcutaneous administration.

132. A binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for use as a medicament.

133. A binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for use in the treatment of cancer.

134. A binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for use in the treatment of an inflammatory disease and/or autoimmune disorder.

135. The binding protein or pharmaceutical composition for use of claim 133, wherein the tumor-associated antigen is CD20, and the cancer is a B-cell malignancy.

136. The binding protein or pharmaceutical composition for use of claim 135, wherein the B-cell malignancy is non-Hodgkin's lymphoma (NHL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Burkett lymphoma, primary mediastinal large B cell lymphoma (PMBCL) or small lymphocytic lymphoma (SLL).

137. The binding protein or pharmaceutical composition for use of claim 133, wherein the tumor-associated antigen is Glypican-3 (GPC3), and the cancer is liver cancer or hepatocellular carcinoma (HCC), non-small cell lung cancer (NSCLC), squamous non-small cell lung cancer (sqNSCLC), ovarian cancer, clear cell ovarian cancer, carcinoma, Merkel cell carcinoma, gastric cancer, hepatoblastoma, or nephroblastoma.

138. The binding protein or pharmaceutical composition for use of claim 133, wherein the tumor-associated antigen is leucine rich repeat containing 15 (LRRC15) and the cancer is melanoma, sarcoma, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial cell carcinoma, osteosarcoma, glioblastoma, lung cancer, non-small cell lung cancer, or thyroid cancer.

139. The binding protein or pharmaceutical composition for use of claim 133, wherein the tumor-associated antigen is BCMA, and the cancer is multiple myeloma.

140. The binding protein or pharmaceutical composition for use of claim 133, wherein the tumor-associated antigen is STEAP2, and the cancer is prostate cancer or Ewing's sarcoma.

141. The binding protein or pharmaceutical composition for use of claim 134, wherein the inflammatory disease and/or autoimmune disorder is scleroderma, systemic lupus erythematosus (SLE), myositis, rheumatoid arthritis (RA), Sjogren's syndrome, or ANCA vasculitis.

142. A binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for use in the treatment of systemic lupus erythematosus (SLE).

143. A binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for use in the treatment of myositis.

144. A binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for use in the treatment of scleroderma.

145. A binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for use in the treatment of rheumatoid arthritis (RA).

146. A binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for use in the treatment of Sjogren's syndrome.

147. A binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for use in the treatment of ANCA vasculitis.

148. A binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for use in the treatment of a B-cell malignancy.

149. A binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for use in the treatment of hepatocellular carcinoma.

150. A binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for use in the treatment of osteosarcoma.

151. A binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for use in the treatment of multiple myeloma.

152. Use of a binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for the manufacture of a medicament for treating cancer.

153. Use of a binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for the manufacture of a medicament for treating an inflammatory and/or autoimmune disorder.

154. The use of claim 152, wherein the cancer is a B-cell malignancy, liver cancer, or HCC.

155. The use of claim 154, wherein the B-cell malignancy is non-Hodgkin's lymphoma (NHL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Burkett lymphoma, primary mediastinal large B cell lymphoma (PMBCL) or small lymphocytic lymphoma (SLL).

156. The use of claim 152, wherein the cancer is melanoma, sarcoma, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial cell carcinoma, osteosarcoma, glioblastoma, lung cancer, non-small cell lung cancer, or thyroid cancer.

157. The use of claim 153, wherein the inflammatory and/or autoimmune disorder is scleroderma, systemic lupus erythematosus (SLE), myositis, rheumatoid arthritis (RA), Sjogren's syndrome, or ANCA vasculitis.

158. Use of a binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for the manufacture of a medicament for the treatment of systemic lupus erythematosus (SLE).

159. Use of a binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for the manufacture of a medicament for the treatment of myositis.

160. Use of a binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for the manufacture of a medicament for the treatment of scleroderma.

161. Use of a binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for the manufacture of a medicament for the treatment of rheumatoid arthritis (RA).

162. Use of a binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for the manufacture of a medicament for the treatment of Sjogren's syndrome.

163. Use of a binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for the manufacture of a medicament for the treatment of ANCA vasculitis.

164. Use of a binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for the manufacture of a medicament for the treatment of a B-cell malignancy.

165. Use of a binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for the manufacture of a medicament for the treatment of hepatocellular carcinoma.

166. Use of a binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for the manufacture of a medicament for the treatment of osteosarcoma.

167. Use of a binding protein of any one of claims 1-89, or the pharmaceutical composition of claim 90, for the manufacture of a medicament for the treatment of multiple myeloma.

168. A binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, and SEQ ID NO: 140 respectively.

169. The binding protein of claim 168, comprising a VH chain according to SEQ ID NO: 141 and a VL according to SEQ ID NO: 142.

170. A binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 147, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, and SEQ ID NO: 140 respectively.

171. The binding protein of claim 170, comprising a VH chain according to SEQ ID NO: 148 and a VL according to SEQ ID NO: 142.

172. A binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 152, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, and SEQ ID NO: 140 respectively.

173. The binding protein of claim 172, comprising a VH chain according to SEQ ID NO: 153 and a VL according to SEQ ID NO: 142.

174. A binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 157, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 158, and SEQ ID NO: 140 respectively.

175. The binding protein of claim 174, comprising a VH chain according to SEQ ID NO: 159 and a VL according to SEQ ID NO: 160.

176. A binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequences of SEQ ID NO: 135, SEQ ID NO: 157, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 165, and SEQ ID NO: 140 respectively.

177. The binding protein of claim 176, comprising a VH chain according to SEQ ID NO: 159 and a VL according to SEQ ID NO: 166.

178. A binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2) and a heavy chain CDR3 (HCDR3) comprising the amino acid sequences of SEQ ID NO: 169, SEQ ID NO: 170 and SEQ ID NO: 171 respectively.

179. The binding protein of claim 178, comprising a VHH chain according to SEQ ID NO: 168.

180. A binding protein comprising a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2) and a heavy chain CDR3 (HCDR3) comprising the amino acid sequences of SEQ ID NO: 62, SEQ ID NO: 40 and SEQ ID NO: 41 respectively.

181. The binding protein of claim 180, comprising a VHH chain according to SEQ ID NO: 43 or SEQ ID NO: 45.

182. A binding protein, comprising a VHH chain according to SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 47 or SEQ ID NO: 48.