EP4731674A2 - Compositions and methods for binding to covalent peptide conjugates - Google Patents

Compositions and methods for binding to covalent peptide conjugates

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Publication number
EP4731674A2
EP4731674A2 EP24826684.3A EP24826684A EP4731674A2 EP 4731674 A2 EP4731674 A2 EP 4731674A2 EP 24826684 A EP24826684 A EP 24826684A EP 4731674 A2 EP4731674 A2 EP 4731674A2
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sequence
seq
composition
amino acid
antigen
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German (de)
French (fr)
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Shohei Koide
Lorenzo MASO
Benjamin G. Neel
Akiko Koide
Epsa RAJAK
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New York University NYU
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New York University NYU
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/70539—MHC-molecules, e.g. HLA-molecules
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/82—Translation products from oncogenes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2833—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against MHC-molecules, e.g. HLA-molecules
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/32—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/40—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against enzymes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/32—Immunoglobulins specific features characterized by aspects of specificity or valency specific for a neo-epitope on a complex, e.g. antibody-antigen or ligand-receptor
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biochemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Zoology (AREA)
  • Oncology (AREA)
  • Cell Biology (AREA)
  • Peptides Or Proteins (AREA)

Abstract

Provided are compositions and methods that include binding partners that specifically bind to peptide conjugate/MHC complex, wherein the peptide conjugate/MHC complex comprises a peptide conjugate that is formed by the covalent reaction of a targeted covalent inhibitor with a peptide. The binding partners are provided as antibodies or antibody derivatives that specifically bind to the peptide conjugate/MHC complexes described herein.

Description

COMPOSITIONS AND METHODS FOR BINDING TO COVALENT PEPTIDE CONJUGATES
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims priority to and benefit of U.S. Provisional Patent Application No. 63/509,472, filed on June 21, 2023, and U.S. Provisional Patent Application No. 63/637,558, filed on April 23, 2024, each of which is entirely incorporated herein by reference.
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH
This invention was made with government support under CAO 16087 and CA267362 awarded by the National Institutes of Health. The government has certain rights in the invention.
BACKGROUND
There is an ongoing and unmet need for agents that can bind to targets that include drugs that are covalently bound to proteins or peptides. In particular, there is a need to improve the efficacy of targeted therapy and also to increase tumor immunogenicity and the efficacy of immune therapy against cancer driven by intracellular oncogenes and loss of tumor suppressor genes. The disclosure is pertinent to these needs.
BRIEF SUMMARY
The present disclosure provides compositions and methods that include binding partners that bind with specificity to target sites on proteins or peptides that comprise a covalently attached molecule. The disclosure illustrates this approach using binding partners in the form of numerous antibodies and antibody derivatives that specifically bind to proteins and peptides that have been covalently modified by attachment of a molecule, wherein the molecules are illustrated by a variety of drugs. Further, the disclosure demonstrates binding partners that bind with specificity to peptides that have been covalently modified by attachment of a small molecule drug are specific for the described covalently modified peptides when presented in the context of a human leukocyte antigen (HLA), wherein HLA is a representative example of a major histocompatibility complex (MHC). Thus, binding partners that are specific for peptide-drug conjugates in an HLA complex are demonstrated. The disclosure also provides binding partners having specificity to two different drug- peptide/MHC complexes. The disclosure includes polynucleotides encoding the described binding partners and cells that are modified to express the binding partners. The disclosure includes diagnostic, prophylactic and therapeutic approaches using the binding partners.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain binds to (i) a first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 100 nM and (ii) a second peptide conjugate/MHC complex with a KD of at most about 100 nM, wherein the first peptide conjugate/MHC complex comprises: (a) a first peptide conjugate, wherein the first peptide conjugate of the first peptide conjugate/MHC complex is a first peptide covalently linked to a first targeted covalent inhibitor or fragment thereof; and (b) a first MHC; wherein the second peptide conjugate/MHC complex comprises: (a) a second peptide conjugate, wherein the second peptide conjugate of the second peptide conjugate/MHC complex is a second peptide covalently linked to a second targeted covalent inhibitor or fragment thereof; and (b) a second MHC; and wherein the first targeted covalent inhibitor is different from the second targeted covalent inhibitor.
In some embodiments, the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 50 nM and/or (ii) the second peptide conjugate/MHC complex with a KD of at most about 50 nM. In some embodiments, the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 10 nM and/or (ii) the second peptide conjugate/MHC complex with a KD of at most about 10 nM. In some embodiments, the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 5 nM and/or (ii) the second peptide conjugate/MHC complex with a KD of at most about 5 nM.
In some embodiments, the first targeted covalent inhibitor is a covalent inhibitor of a RAS protein and the second targeted covalent inhibitor is a covalent inhibitor of a RAS protein. In some embodiments, the first targeted covalent inhibitor is a covalent inhibitor of KRASG12C and the second targeted covalent inhibitor is a covalent inhibitor of KRASG12C. In some embodiments, (i) the first targeted covalent inhibitor is adagrasib and the second targeted covalent inhibitor is sotorasib, (ii) the first targeted covalent inhibitor is adagrasib and the second targeted covalent inhibitor is divarasib; or (iii) the first targeted covalent inhibitor is divarasib and the second targeted covalent inhibitor is sotorasib.
In some embodiments, the antigen-binding domain (i) does not bind to a free first targeted covalent inhibitor that is not conjugated to a peptide with a dissociation constant (KD) of less than 200 nM, (ii) does not bind to the first peptide with a KD of less than 200 nM, (iii) does not bind to a free first peptide conjugate that is not complexed with an MHC with a KD of less than 200 nM, or (iv) any combination above.
In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex with a greater affinity than to the first peptide, the free first targeted covalent inhibitor, and/or the free first peptide conjugate. In some embodiments, the affinity of the antigen-binding domain for the first peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the antigen-binding domain for the first peptide or the free first targeted covalent inhibitor. In some embodiments, the antigen-binding domain does not detectably bind to a complex of the first peptide with the first MHC, wherein the first peptide is not covalently bound to the first targeted covalent inhibitor or fragment thereof. In some embodiments, the antigen-binding domain does not detectably bind to the free first targeted covalent inhibitor. In some embodiments, the antigen-binding domain binds to a free first peptide conjugate with a dissociation constant (KD) that is at least 2.5 times more than a KD of the antibody or the antigen-binding fragment to the first peptide conjugate/MHC complex.
In some embodiments, the antigen-binding domain (i) does not bind to a free second targeted covalent inhibitor that is not conjugated to a peptide with a dissociation constant (KD) of less than 200 nM, (ii) does not bind to the second peptide with a KD of less than 200 nM, (iii) does not bind to a free second peptide conjugate that is not complexed with an MHC with a KD of less than 200 nM, or (iv) any combination above.
In some embodiments, the antigen-binding domain binds to the second peptide conjugate/MHC complex with a greater affinity than to the second peptide, the free second targeted covalent inhibitor, and/or the free second peptide conjugate.
In some embodiments, the affinity of the antigen-binding domain for the second peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the antigenbinding domain for the second peptide or the free second targeted covalent inhibitor. In some embodiments, the antigen-binding domain does not detectably bind to a complex of the second peptide with the second MHC, wherein the second peptide is not covalently bound to the second targeted covalent inhibitor or fragment thereof. In some embodiments, the antigen-binding domain does not detectably bind to the free second targeted covalent inhibitor. In some embodiments, the antigen-binding domain binds to a free second peptide conjugate with a dissociation constant KD that is at least 2.5 times more than a KD of the antibody or the antigen-binding fragment to the second peptide conjugate/MHC complex.
In some embodiments, the first peptide and the second peptide comprise the same amino acid sequence. In some embodiments, the first peptide and the second peptide consist of the same amino acid sequence. In some embodiments, the first peptide or the second peptide comprises the amino acid sequence of VVVGACGVGK. In some embodiments, the first peptide and second peptide comprise a different amino acid sequence. In some embodiments, the first peptide and the second peptide comprise a shared contiguous amino acid sequence that is at least 3, 4, 5, 6, 7, 8, or 9 amino acids in length. In some embodiments, the first peptide and/or the second peptide comprises an amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV. In some embodiments, the first MHC or the second MHC is encoded by an HLA, wherein the HLA is HLA-A*02:01, HLA- A*03:01, or HLA-A* 11:01.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain binds to a peptide conjugate/MHC complex, wherein the peptide conjugate of the peptide conjugate/MHC complex is a peptide covalently linked to a targeted covalent inhibitor or fragment thereof and a MHC; and wherein the antigen-binding domain binds to the peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 50 nM; wherein the targeted covalent inhibitor is adagrasib.
In some embodiments, the peptide conjugate/MHC complex is a first peptide conjugate/MHC complex comprising a first peptide conjugate comprising a first peptide and a first targeted covalent inhibitor or fragment thereof; and wherein the antigen-binding domain further binds to a second peptide conjugate/MHC complex comprising (a) a second peptide conjugate, wherein the second peptide conjugate of the second peptide conjugate/MHC complex is a second peptide covalently linked to a second targeted covalent inhibitor or fragment thereof; and (b) a second MHC.
In some embodiments, the antigen-binding domain binds to the second peptide conjugate/MHC complex with a KD of at most about 50 nM. In some embodiments, the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 10 nM and (ii) the second peptide conjugate/MHC complex with a KD of at most about 10 nM. In some embodiments, the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 5 nM and (ii) the second peptide conjugate/MHC complex with a KD of at most about 5 nM.
In some embodiments, the second targeted covalent inhibitor is sotorasib or divarasib. In some embodiments, the antigen-binding domain does not bind to a free first targeted covalent inhibitor with a dissociation constant (KD) of less than 200 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex with a greater affinity than to the first peptide or the free first targeted covalent inhibitor. In some embodiments, the affinity of the antigen-binding domain for the first peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the antigen-binding domain for the first peptide or the free first targeted covalent inhibitor.
In some embodiments, the antigen-binding domain does not detectably bind to a complex of the first peptide with a first MHC, wherein the first peptide is not covalently bound to the first targeted covalent inhibitor or fragment thereof. In some embodiments, the antigen-binding domain does not detectably bind to the free first targeted covalent inhibitor. In some embodiments, the antigen-binding domain binds to a free first peptide conjugate with a dissociation constant KD) that is at least 2.5 times more than a Ko of the antibody or the antigen-binding fragment to the first peptide conjugate/MHC complex.
In some embodiments, the antigen-binding domain does not bind to a free second targeted covalent inhibitor with a dissociation constant (KD) of less than 200 nM. In some embodiments, the antigen-binding domain binds to the second peptide conjugate/MHC complex with a greater affinity than to the second peptide or the free second targeted covalent inhibitor. In some embodiments, the affinity of the antigen-binding domain for the second peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the antigenbinding domain for the second peptide or the free second targeted covalent inhibitor.
In some embodiments, the antigen-binding domain does not detectably bind to a complex of the second peptide with the second MHC, wherein the second peptide is not covalently bound to the second targeted covalent inhibitor or fragment thereof. In some embodiments, the antigen-binding domain does not detectably bind to the free second targeted covalent inhibitor. In some embodiments, the antigen-binding domain binds to a free second peptide conjugate with a dissociation constant KD) that is at least 2.5 times more than a Ko of the antibody or the antigen-binding fragment to the second peptide conjugate/MHC complex.
In some embodiments, the first peptide and the second peptide comprise the same amino acid sequence. In some embodiments, the first peptide and the second peptide consist of the same amino acid sequence. In some embodiments, the first peptide or the second peptide comprises the amino acid sequence of VVVGACGVGK. In some embodiments, the first peptide and second peptide comprise a different amino acid sequence. In some embodiments, the first peptide or the second peptide comprises an amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV. In some embodiments, the MHC or the second MHC is an HL A, and wherein the HL A is HLA-A*02:01, HLA-A*03:01, or HLA-A* 11 :01.
In some embodiments, the first peptide or the second peptide comprises a nucleophilic or an electrophilic residue, wherein the residue comprises cysteine, aspartic acid, arginine, serine, or tyrosine. In some embodiments, the first peptide or the second peptide comprises a cysteine residue. In some embodiments, the first peptide conjugate is formed by a covalent reaction between the first targeted covalent inhibitor and a cysteine residue in the first peptide. In some embodiments, the second peptide conjugate is formed by a covalent reaction between the second targeted covalent inhibitor and a cysteine residue in the second peptide.
In some embodiments, the first peptide or the second peptide is a segment of a protein that is associated with a cancer, optionally wherein the protein is encoded by a gene that is mutated in a cancer. In some embodiments, the first peptide or the second peptide is a segment of an enzyme, and wherein the first targeted covalent inhibitor or second targeted covalent inhibitor is an inhibitor of the enzyme. In some embodiments, the enzyme is a kinase or GTPase.
In some embodiments, the first peptide or the second peptide is or is derived from KRAS. In some embodiments, the first peptide or the second peptide comprises a segment of KRASG12C, KRASG12D, or KRASG12R. In some embodiments, the first targeted covalent inhibitor or the second targeted covalent inhibitor is (i) a tri-complex KRASG12C inhibitor or a KRASG12C degrader, (ii) a tri-complex KRASG12D inhibitor or a KRASG12D degrader, or (iii) a tri-complex KRASG12R inhibitor or a KRASG12R degrader.
In some embodiments, the first peptide conjugate or the second peptide conjugate comprises a compound selected from the group consisting of compounds (1), (2), and (3):
wherein the compound is covalently bonded to a cysteine residue in a peptide, wherein the peptide comprises an amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV. In some embodiments, the peptide comprises the amino acid sequence of VVVGACGVGK or VVGACGVGK and the MHC is HLA-A*03 :01 or HLA-A* 11 :01. In some embodiments, the peptide comprises the amino acid sequence of KLVVVGACGV and the MHC is HLA-A*02:01. In some embodiments, the first peptide conjugate is formed by the covalent reaction of adagrasib with a KRASG12C peptide. In some embodiments, the second peptide conjugate is formed by the covalent reaction of sotorasib with a KRASG12C peptide. In some embodiments, the second peptide conjugate is formed by the covalent reaction of divarasib with a KRASG12C peptide.
In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of X1SX3X4SIH, wherein Xi is I, F or V, X3 is S or Y, and XHs S or Y; (ii) a HC CDR2 comprising an amino acid sequence of X1ISX4X5X6X7X8TX10YADSVKG, wherein Xi is S or Y, X4 is S or P, X5 is S or Y, X6 is S or Y, X7 is S or G, Xs is S or Y, and X10 is S or Y, and (iii) a HC CDR3 comprising an amino acid sequence of XNX1X2DY, wherein XN IS an amino acid sequence selected from LWAS, FQWY, GYGW, GWYYL, YWYYM, YWYYL, GYYYPYY, SYYGFWQALW, SSRQYYHSQVEPPM, SGYYSSHWYLQSWYQ, and HYSEKWWGWYTMYID, Xi is G or A, and X2 is L or M; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and (iii) a LC CDR3 comprising the amino acid sequence of QQX3X4X5X6X7X8X9X10T, wherein X3 is W, T, S, A or G, X4is N, W, S, G, Y, D or K, X5 is W, Y, S, A or T, X6 is G, S, Y, L, W, E or D, X7 is W, S, H, Y, E, F or absent, Xs is P, Q, S, W, E, L, G or absent, X9 is L or P, and X10 is I, L, F, or V.
In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of LWASGLDY; and/or the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4is G, L, S, E or Y, Xs is W S, Y, E, or F, Xe is P, W, G, E, or Q, and X7 is I, F, or V.
In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising an amino acid sequence of SYYGFWQALWALDY; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V. In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising an amino acid sequence of GYYYPYYAMDY; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising an amino acid sequence of GYYYPYYAMDY; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4is G, L, S, E or Y, Xs is W S, Y, E, or F, Xe is P, W, G, E, or Q, and X7 is I, F, or V.
In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising an amino acid sequence of X1X2X3X4X5MDY, wherein Xi is F or G, X2 is Q or Y, X3 is W or G, X4 is Y or W, and X5 is A or G; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4is G, L, S, E or Y, Xs is W S, Y, E, or F, Xe is P, W, G, E, or Q, and X7 is I, F, or V.
In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising an amino acid sequence of SGYYSSHWYLQSWYQAMDY; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising an amino acid sequence of SGYYSSHWYLQSWYQAMDY; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQXIX2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising an amino acid sequence of HYSEKWWGWYTMYIDAMDY; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising an amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of SSRQYYHSQVEPPMAMDY; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In some embodiments, the antigen-binding domain comprises the HC CDR1, HC CDR2, and HC CDR3 amino acid sequences of a VH amino acid sequence and/or the LC CDR1, LC CDR2, and LC CDR3 amino acid sequences of a VL amino acid sequence of a polypeptide selected from the group consisting of RM_001, RM_002, RM_003, RM_004, RM_005, RM_006, RM_007, RM_008, RM_009, RM_010, RM_011, RM_012, RM_013, RM_014, RM_015, RM_016, RM_017, RM_018, RM_019, RM_020, RM_021, RM_022, , RM 023, RM_024, RM_025, RM_026, RM_027, RM_028 , RM_029, RM_030, RM_031, and RM 032; or a variant thereof comprising 1-5 amino acid changes in one or more of the CDR amino acid sequences.
In some embodiments, the antigen-binding domain comprises the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and/or LC CDR3 amino acid sequences of a polypeptide selected from the group consisting of RM_001, RM_002, RM_003, RM_004, RM_005, RM_006, RM_007, RM_008, RM_009, RM_010, RM_011, RM_012, RM_013, RM_014, RM_015, RM_016, RM_017, RM_018, RM_019, RM_020, RM_021, RM_022, , RM 023, RM_024, RM_025, RM_026, RM_027, RM_028 , RM_029, RM_030, RM_031, and RM 032, or a variant thereof comprising 1-5 amino acid changes in one or more of the CDR amino acid sequences.
In some embodiments, the antigen-binding domain comprises a VH and/or a VL amino acid sequence that is 90%, 95%, or 100% identical to the following VH and VL sequences:
RM_001
VL :DIQMTQ SP S SLS AS VGDRVTITCRASQ S VS S AV AW YQQKPGKAPKLLI YS AS SL YSGVPSRFSGSRSGTDFTL
TISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV
VH:EVQLVESGGGL VQPGGSLRLSC AASGFTIS S S SIHWVRQAPGKGLEWVASIS S Y YGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWG QGTLVTVSS
RM_002
VL :DIQMTQ SP S SLS AS VGDRVTITCRASQ S VS S AV AW YQQKPGKAPKLLI YS AS SL YSGVPSRFSGSRSGTDFTL
TISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV
VH:EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS SY YGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWG QGTLVTVSS
RM_003
VL :DIQMTQ SP S SLS AS VGDRVTITCRASQ S VS S AV AW YQQKPGKAPKLLI YS AS SL YSGVPSRFSGSRSGTDFTL
TISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRTV
VH:EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S S SGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALW ALD YWGQGTL VT VS S
RM_004
VL :DIQMTQ SP S SLS AS VGDRVTITCRASQ S VS S AV AW YQQKPGKAPKLLI YS AS SL YSGVPSRFSGSRSGTDFTL
TISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISSS SGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALW ALD YWGQGTL VT VS S
RM_005
VL :DIQMTQ SP S SLS AS VGDRVTITCRASQ S VS S AV AW YQQKPGKAPKLLI YS AS SL YSGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISS YYGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAM D YWGQGTL VTVSS
RM_006 VL :DIQMTQ SP S SLS AS VGDRVTITCRASQ S VS S AV AW YQQKPGKAPKLLI YS AS SL YSGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISP YYGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAM DYWGQGTLVTVSS
RM_007
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQTSWYHSLITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSY
YGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDY WGQGTLVTVSS
RM_008
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVASISSY YGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDY WGQGTLVTVSS
RM_009
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSY
YGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDY WGQGTLVTVSS
RM_010
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSWLYWLVTFGQGTKVEIKRTV VH:EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SS GSTSYADSVKGRFTISAD
TSKNTAYLQMNSLRAEDTAVYYCARFQWYAMDYWGQGTLVTVSS
RM_011
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTISSYSIHWVRQAPGKGLEWVAYISSYS GYTSYADSVKGRFTISAD
TSKNTAYLQMNSLRAEDTAVYYCARGYGWGMDYWGQGTLVTVSS
RM_012
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTISYSSIHWVRQAPGKGLEWVAYISSSS GYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQS W YQ AMD YWGQGTL VT VS S RM_013
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQASYGPITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSY
YGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQS W YQ AMD YWGQGTL VT VS S
RM_014
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQWWSSSQLITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVASISSY
YGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARHYSEKWWGWY TMYID AMD YWGQGTL VT VS S
RM_015
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPY
SGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYW
GQGTLVTVSS
RM_016
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWG
QGTLVTVSS
RM_017
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPY
SGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYW
GQGTLVTVSS
RM_018
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPY
SGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYW GQGTLVTVSS
RM_019
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWG QGTLVTVSS
RM_020
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPY
SGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYW
GQGTLVTVSS
RM_021
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG
QGTLVTVSS
RM_022
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG
QGTLVTVSS
RM_023
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSYYEELITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG
QGTLVTVSS
RM_024
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQAYSDPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG
QGTLVTVSS
RM 025
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG
QGTLVTVSS
RM_026 VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSGSYLLITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG QGTLVTVSS
RM_027
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQADYEFGLITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG QGTLVTVSS
RM_028
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVAYISSSY GYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPP MAMD YWGQGTL VT VS S
RM_029
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVAYISSS YGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEP PMAMD YWGQGTL VT VS S
RM_030
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYISSSY GYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPP MAMD YWGQGTL VT VS S
RM_031
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSC AASGFT VS YS SIHWVRQAPGKGLEWVAYIS S S YGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEP PMAMD YWGQGTL VT VS S
RM_032
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPY SGYTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYLGMDYW GQGTLVTVSS In some embodiments, the polypeptide specifically binds to the first peptide conjugate/MHC complex and a T cell antigen. In some embodiments, the polypeptide specifically binds to the first peptide conjugate/MHC complex and human CD3. In some embodiments, the polypeptide specifically binds to the second peptide conjugate/MHC complex and a T cell antigen. In some embodiments, the polypeptide specifically binds to the second peptide conjugate/MHC complex and human CD3. In some embodiments, the polypeptide comprises a heavy chain constant region selected from the group consisting of human IgM, IgGi, IgG2, IgGs, IgG4, IgAi, and IgA. In some embodiments, the heavy chain constant region comprises one or more amino acid substitutions, deletions, or additions in the Fc region. In some embodiments, the polypeptide is conjugated to a detectable label, a chemotherapeutic agent, a radioisotope, or a toxin.
In some embodiments, the polypeptide is comprised within a chimeric antigen receptor. In some embodiments, the polypeptide is expressed by a T cell, a killer macrophage, a neutrophil or a natural killer cell.
In an aspect, the present disclosure provides a polynucleotide encoding the polypeptide described herein.
In an aspect, the present disclosure provides a polynucleotide encoding a heavy chain variable region and/or a light chain variable region of the polypeptide described herein. In some embodiments, the polynucleotide is a modified polynucleotide.
In an aspect, the present disclosure provides a vector comprising the polynucleotide described herein. In some embodiments, the vector is a viral vector. In some embodiments, the viral vector is an adenoviral vector, lentiviral vector, retroviral vector, or adeno- associated viral vector.
In an aspect, the present disclosure provides a recombinant host cell comprising: (a) the polynucleotide described herein; (b) the vector described herein; (c) a first polynucleotide encoding a VH or a heavy chain of the polypeptide described herein, and a second polynucleotide encoding a VL or a light chain of the polypeptide described herein; or (d) a first vector comprising a first polynucleotide encoding a VH or a heavy chain of the polypeptide described herein, and a second vector comprising a second polynucleotide encoding a VL or a light chain of the polypeptide described herein.
In an aspect, the present disclosure provides a pharmaceutical composition comprising the polypeptide described herein, the polynucleotide described herein, the vector described herein, or the host cell described herein and a pharmaceutically acceptable carrier or excipient. In an aspect, the present disclosure provides a method of producing a polypeptide, the method comprising culturing the host cell described herein under suitable conditions so that the polynucleotide is expressed and the binding partner is produced.
In an aspect, the present disclosure provides an eukaryotic cell comprising the polynucleotide described herein or the vector described herein, wherein the cell is optionally a totipotent, multipotent, or pluripotent stem cell, wherein optionally the stem cell has an induced stem cell phenotype, or wherein the cell is optionally a leukocyte, optionally a CD4+ T cell, optionally a CD8+ T cell, optionally a y5 T cell, optionally a natural killer cell , a neutrophil or a macrophage.
In an aspect, the present disclosure provides a method comprising administering to an individual in need thereof the polypeptide described herein, the polynucleotide described herein, the vector described herein, the pharmaceutical composition described herein, or the cell described herein.
In an aspect, the present disclosure provides a cell free peptide conjugate/MHC complex comprising: (a) an isolated peptide conjugate formed by the covalent reaction of a targeted covalent inhibitor with a peptide; and (b) an MHC, wherein the targeted covalent inhibitor is sotorasib, adagrasib, or divarasib, wherein the MHC is an HLA, and wherein the HL A i s HL A- A* 02 : 01 , HL A- A* 03 : 01 , or HL A- A* 11 :01.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR3 comprising an amino acid sequence of XNX1X2DY, wherein X\ is an amino acid sequence selected from LWAS, FQWY, GYGW, GWYYL, YWYYM, YWYYL, GYYYPYY, SYYGFWQALW, SSRQYYHSQVEPPM, SGYYSSHWYLQSWYQ, and HYSEKWWGWYTMYID, Xi is G or A, and X2 is L or M; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and (iii) a LC CDR3 comprising the amino acid sequence of QQX3X4X5X6X7X8X9X10T, wherein X3 is W, T, S, A or G, X4is N, W, S, G, Y, D or K, X5 is W, Y, S, A or T, X6 is G, S, Y, L, W, E or D, X7 is W, S, H, Y, E, F or absent, X8 is P, Q, S, W, E, L, G or absent, X9 is L or P, and X10 is I, L, F, or V. In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of X1SX3X4SIH, wherein Xi is I, F or V, X3 is S or Y, and X4 is S or Y. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of X1ISX4X5X6X7X8TX10YADSVKG, wherein Xi is S or Y, X4 is S or P, X5 is S or Y, Xe is S or Y, X7 is S or G, Xs is S or Y, and X10 is S or Y.
In some embodiments, the VL comprises a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of X1SX3X4SIH, wherein Xi is I, F or V, X3 is S or Y, and X4 is S or Y; (ii) a HC CDR2 comprising an amino acid sequence of X1ISX4X5X6X7X8TX10YADSVKG, wherein Xi is S or Y, X4is S or P, X5 is S or Y, Xe is S or Y, X7 is S or G, Xs is S or Y, and X10 is S or Y, and (iii) a HC CDR3 comprising an amino acid sequence of XNX1X2DY, wherein XN IS an amino acid sequence selected from LWAS, FQWY, GYGW, GWYYL, YWYYM, YWYYL, GYYYPYY, SYYGFWQALW, SSRQYYHSQVEPPM, SGYYSSHWYLQSWYQ, and HYSEKWWGWYTMYID, Xi is G or A, and X2 is L or M; and/or (b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and (iii) a LC CDR3 comprising the amino acid sequence of QQX3X4X5X6X7X8X9X10T, wherein X3 is W, T, S, A or G, X4is N, W, S, G, Y, D or K, X5 is W, Y, S, A or T, X6 is G, S, Y, L, W, E or D, X7 is W, S, H, Y, E, F or absent, Xs is P, Q, S, W, E, L, G or absent, X9 is L or P, andXio is I, L, F, or V.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of LWASGLDY (SEQ ID NO: 5).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSYYGSTSYADSVKG (SEQ ID NO: 4). In some embodiments, the VH comprises a HC CDR1 sequence of ISSSSIH (SEQ ID NO: 3). In some embodiments, the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQWNWGWPLIT (SEQ ID NO: 8). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 7). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 6). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of ISSSSIH (SEQ ID NO: 3), a HC CDR2 sequence of SISSYYGSTSYADSVKG (SEQ ID NO: 4), a HC CDR3 sequence of LWASGLDY (SEQ ID NO: 5), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 6), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 7), and a LC CDR3 sequence of QQWNWGWPLIT (SEQ ID NO: 8). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTIS S S SMWVRQAPGKGLEWVASIS S YYGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWGQGTLV TVSS (SEQ ID NO: 2). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV (SEQ ID NO: 1).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of LWASGLDY (SEQ ID NO: 13).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSYYGSTSYADSVKG (SEQ ID NO: 12). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 11). In some embodiments, the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQWNWGWPLIT (SEQ ID NO: 16). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 15). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 14). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 11), a HC CDR2 sequence of SISSYYGSTSYADSVKG (SEQ ID NO: 12), a HC CDR3 sequence of LWASGLDY (SEQ ID NO: 13), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 14), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 15), and a LC CDR3 sequence of QQWNWGWPLIT (SEQ ID NO: 16). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S YYGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWGQGTLV TVSS (SEQ ID NO: 10). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV (SEQ ID NO: 9).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SYYGFWQALWALDY (SEQ ID NO: 21).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 20). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 19). In some embodiments, the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQWWYGSPLFT (SEQ ID NO: 24). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 23). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 22). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 19), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 20), a HC CDR3 sequence of SYYGFWQALWALDY (SEQ ID NO: 21), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 22), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 23), and a LC CDR3 sequence of QQWWYGSPLFT (SEQ ID NO: 24). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALWALDYW GQGTLVTVSS (SEQ ID NO: 18). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRT (SEQ ID NO: 17).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SYYGFWQALWALDY (SEQ ID NO: 29).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 28). In some embodiments, the VH comprises a HC CDR1 sequence of FSSYSIH (SEQ ID NO: 27). In some embodiments, the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQWWYGSPLFT (SEQ ID NO: 32). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 31). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 30). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSYSIH (SEQ ID NO: 27), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 28), a HC CDR3 sequence of SYYGFWQALWALDY (SEQ ID NO: 29), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 30), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 31), and a LC CDR3 sequence of QQWWYGSPLFT (SEQ ID NO: 32). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S YSIHWVRQAPGKGLEW VASIS S S SGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALWALDYW GQGTLVTVSS (SEQ ID NO: 26). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRT (SEQ ID NO: 25).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GYYYP YYAMD Y (SEQ ID NO: 37).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 36). In some embodiments, the VH comprises a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 35). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGKTYYPIT (SEQ ID NO: 40). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 39). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 38). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 35), a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 36), a HC CDR3 sequence of GYYYP YYAMD Y (SEQ ID NO: 37), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 38), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 39), and a LC CDR3 sequence of QQGKTYYPIT (SEQ ID NO: 40). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYP YYAMD YWG QGTLVTVSS (SEQ ID NO: 34). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 33).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GYYYP YYAMD Y (SEQ ID NO: 45).
In some embodiments, the VH comprises a HC CDR2 sequence of SISPYYGSTYYADSVKG (SEQ ID NO: 44). In some embodiments, the VH comprises a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 43). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGKTYYPIT (SEQ ID NO: 48). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 47). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 46). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 43), a HC CDR2 sequence of SISPYYGSTYYADSVKG (SEQ ID NO: 44), a HC CDR3 sequence of GYYYP YYAMD Y (SEQ ID NO: 45), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 46), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 47), and a LC CDR3 sequence of QQGKTYYPIT (SEQ ID NO: 48). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF SYYSIHWVRQAPGKGLEWVASISP YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYP YYAMD YWG QGTLVTVSS (SEQ ID NO: 42). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 41).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GYYYP YYAMD Y (SEQ ID NO: 53).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 52). In some embodiments, the VH comprises a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 51). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQTSWYHSLIT (SEQ ID NO: 56). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 55). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 54). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 51), a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 52), a HC CDR3 sequence of GYYYP YYAMD Y (SEQ ID NO: 53), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 54), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 55), and a LC CDR3 sequence of QQTSWYHSLIT (SEQ ID NO: 56). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYP YYAMD YWG QGTLVTVSS (SEQ ID NO: 50). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQTSWYHSLITFGQGTKVEIKRTV (SEQ ID NO: 49).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GYYYP YYAMD Y (SEQ ID NO: 61). In some embodiments, the VH comprises a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 60). In some embodiments, the VH comprises a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 59). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGKTYYPIT (SEQ ID NO: 64). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 63). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 62). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 59), a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 60), a HC CDR3 sequence of GYYYP YYAMD Y (SEQ ID NO: 61), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 62), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 63), and a LC CDR3 sequence of QQGKTYYPIT (SEQ ID NO: 64). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVASISSYYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYP YYAMD YWG QGTLVTVSS (SEQ ID NO: 58). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 57).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GYYYP YYAMD Y (SEQ ID NO: 69).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 68). In some embodiments, the VH comprises a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 67). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 72). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 71). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 70). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 67), a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 68), a HC CDR3 sequence of GYYYP YYAMD Y (SEQ ID NO: 69), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 70), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 71), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 72). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYP YYAMD YWG QGTLVTVSS (SEQ ID NO: 66). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 65).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of FQWYAMDY (SEQ ID NO: 77).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 76). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 75). In some embodiments, the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWLVT (SEQ ID NO: 80). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 79). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 78). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 75), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 76), a HC CDR3 sequence of FQWYAMDY (SEQ ID NO: 77), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 78), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 79), and a LC CDR3 sequence of QQSSWLYWLVT (SEQ ID NO: 80). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFQWYAMDYWGQGTL VTVSS (SEQ ID NO: 74). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWLVTFGQGTKVEIKRTV (SEQ ID NO: 73).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GYGWGMDY (SEQ ID NO: 85).
In some embodiments, the VH comprises a HC CDR2 sequence of YISSYSGYTSYADSVKG (SEQ ID NO: 84). In some embodiments, the VH comprises a HC CDR1 sequence of ISSYSIH (SEQ ID NO: 83). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQWNWGWPLIT (SEQ ID NO: 88). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 87). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 86). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of ISSYSIH (SEQ ID NO: 83), a HC CDR2 sequence of YISSYSGYTSYADSVKG (SEQ ID NO: 84), a HC CDR3 sequence of GYGWGMDY (SEQ ID NO: 85), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 86), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 87), and a LC CDR3 sequence of QQWNWGWPLIT (SEQ ID NO: 88). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTIS S YSMWVRQAPGKGLEWVAYIS S YSGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYGWGMDYWGQGT LVTVSS (SEQ ID NO: 82). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV (SEQ ID NO: 81).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SGYYSSHWYLQSWYQAMDY (SEQ ID NO: 93).
In some embodiments, the VH comprises a HC CDR2 sequence of YISSSSGYTSYADSVKG (SEQ ID NO: 92). In some embodiments, the VH comprises a HC CDR1 sequence of ISYSSIH (SEQ ID NO: 91). In some embodiments, the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSAWYPVT (SEQ ID NO: 96). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 95). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 94). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of ISYSSIH (SEQ ID NO: 91), a HC CDR2 sequence of YISSSSGYTSYADSVKG (SEQ ID NO: 92), a HC CDR3 sequence of SGYYSSHWYLQSWYQAMDY (SEQ ID NO: 93), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 94), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 95), and a LC CDR3 sequence of QQSSAWYPVT (SEQ ID NO: 96). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTISYSSIHWVRQAPGKGLEWVAYISSSSGYT SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQSWYQ AMDYWGQGTLVTVSS (SEQ ID NO: 90). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV (SEQ ID NO: 89).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SGYYSSHWYLQSWYQAMDY (SEQ ID NO: 101).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 100). In some embodiments, the VH comprises a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 99). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASYGPIT (SEQ ID NO: 104). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 103). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 102). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 99), a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 100), a HC CDR3 sequence of SGYYSSHWYLQSWYQAMDY (SEQ ID NO: 101), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 102), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 103), and a LC CDR3 sequence of QQASYGPIT (SEQ ID NO: 104). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQSWY QAMDYWGQGTLVTVSS (SEQ ID NO: 98). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYGPITFGQGTKVEIKRTV (SEQ ID NO: 97).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of HYSEKWWGWYTMYIDAMDY (SEQ ID NO: 109).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 108). In some embodiments, the VH comprises a HC CDR1 sequence of VSYSSIH (SEQ ID NO: 107). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQWWSSSQLIT (SEQ ID NO: 112). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 111). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 110). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSYSSIH (SEQ ID NO: 107), a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 108), a HC CDR3 sequence of HYSEKWWGWYTMYIDAMDY (SEQ ID NO: 109), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 110), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 111), and a LC CDR3 sequence of QQWWSSSQLIT (SEQ ID NO: 112). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFT VS YSSIHWVRQAPGKGLEWVASISS YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARHYSEKWWGWYTMYI DAMDYWGQGTLVTVSS (SEQ ID NO: 106). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWSSSQLITFGQGTKVEIKRTV (SEQ ID NO: 105).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GWYYLGMDY (SEQ ID NO: 117).
In some embodiments, the VH comprises a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 116). In some embodiments, the VH comprises a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 115). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGKTYYPIT (SEQ ID NO: 120). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 119). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 118). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 115), a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 116), a HC CDR3 sequence of GWYYLGMDY (SEQ ID NO: 117), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 118), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 119), and a LC CDR3 sequence of QQGKTYYPIT (SEQ ID NO: 120). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQ GTLVTVSS (SEQ ID NO: 114). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 113).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GWYYLGMDY (SEQ ID NO: 125).
In some embodiments, the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 124). In some embodiments, the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 123). In some embodiments, the antigen- binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGKTYYPIT (SEQ ID NO: 128). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 127). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 126). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 123), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 124), a HC CDR3 sequence of GWYYLGMDY (SEQ ID NO: 125), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 126), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 127), and a LC CDR3 sequence of QQGKTYYPIT (SEQ ID NO: 128). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQG TLVTVSS (SEQ ID NO: 122). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 121).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GWYYLGMDY (SEQ ID NO: 133).
In some embodiments, the VH comprises a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 132). In some embodiments, the VH comprises a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 131). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 136). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 135). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 134). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 131), a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 132), a HC CDR3 sequence of GWYYLGMDY (SEQ ID NO: 133), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 134), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 135), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 136). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQ GTLVTVSS (SEQ ID NO: 130). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 129).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GWYYLGMDY (SEQ ID NO: 141).
In some embodiments, the VH comprises a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 140). In some embodiments, the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 139). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 144). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 143). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 142). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 139), a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 140), a HC CDR3 sequence of GWYYLGMDY (SEQ ID NO: 141), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 142), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 143), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 144). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPYSGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQG TLVTVSS (SEQ ID NO: 138). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 137). In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GWYYLGMDY (SEQ ID NO: 149).
In some embodiments, the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 148). In some embodiments, the VH comprises a HC CDR1 sequence of FSSYSIH (SEQ ID NO: 147). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 152). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 151). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 150). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSYSIH (SEQ ID NO: 147), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 148), a HC CDR3 sequence of GWYYLGMDY (SEQ ID NO: 149), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 150), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 151), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 152). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISPYSSYT SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGT LVTVSS (SEQ ID NO: 146). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 145).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GWYYLGMDY (SEQ ID NO: 157).
In some embodiments, the VH comprises a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 156). In some embodiments, the VH comprises a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 155). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSAWYPVT (SEQ ID NO: 160). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 159). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 158). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 155), a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 156), a HC CDR3 sequence of GWYYLGMDY (SEQ ID NO: 157), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 158), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 159), and a LC CDR3 sequence of QQSSAWYPVT (SEQ ID NO: 160). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQ GTLVTVSS (SEQ ID NO: 154). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV (SEQ ID NO: 153).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 165).
In some embodiments, the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 164). In some embodiments, the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 163). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 168). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 167). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 166). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 163), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 164), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 165), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 166), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 167), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 168). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 162). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 161).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 173).
In some embodiments, the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 172). In some embodiments, the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 171). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSAWYPVT (SEQ ID NO: 176). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 175). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 174). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 171), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 172), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 173), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 174), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 175), and a LC CDR3 sequence of QQSSAWYPVT (SEQ ID NO: 176). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 170). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV (SEQ ID NO: 169).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 181).
In some embodiments, the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 180). In some embodiments, the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 179). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSYYEELIT (SEQ ID NO: 184). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 183). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 182). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 179), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 180), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 181), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 182), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 183), and a LC CDR3 sequence of QQSSYYEELIT (SEQ ID NO: 184). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 178). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSYYEELITFGQGTKVEIKRTV (SEQ ID NO: 177).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 189).
In some embodiments, the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 188). In some embodiments, the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 187). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQAYSDPLT (SEQ ID NO: 192). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 191). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 190). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 187), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 188), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 189), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 190), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 191), and a LC CDR3 sequence of QQAYSDPLT (SEQ ID NO: 192). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 186). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQAYSDPLTFGQGTKVEIKRTV (SEQ ID NO: 185).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 197).
In some embodiments, the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 196). In some embodiments, the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 195). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGSSSLLT (SEQ ID NO: 200). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 199). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 198). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 195), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 196), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 197), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 198), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 199), and a LC CDR3 sequence of QQGSSSLLT (SEQ ID NO: 200). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 194). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV (SEQ ID NO: 193).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 205).
In some embodiments, the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 204). In some embodiments, the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 203). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSGSYLLIT (SEQ ID NO: 208). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 207). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 206). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 203), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 204), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 205), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 206), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 207), and a LC CDR3 sequence of QQSGSYLLIT (SEQ ID NO: 208). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 202). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSGSYLLITFGQGTKVEIKRTV (SEQ ID NO: 201).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 213). In some embodiments, the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 212). In some embodiments, the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 211). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQADYEFGLIT (SEQ ID NO: 216). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 215). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 214). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 211), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 212), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 213), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 214), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 215), and a LC CDR3 sequence of QQADYEFGLIT (SEQ ID NO: 216). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 210). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQADYEFGLITFGQGTKVEIKRTV (SEQ ID NO: 209).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 221).
In some embodiments, the VH comprises a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 220). In some embodiments, the VH comprises a HC CDR1 sequence of FSSYSIH (SEQ ID NO: 219). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 224). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 223). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 222). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSYSIH (SEQ ID NO: 219), a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 220), a HC CDR3 sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 221), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 222), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 223), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 224). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 218). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 217).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 229).
In some embodiments, the VH comprises a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 228). In some embodiments, the VH comprises a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 227). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 232). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 231). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 230). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 227), a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 228), a HC CDR3 sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 229), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 230), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 231), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 232). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 226). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 225).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 237).
In some embodiments, the VH comprises a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 236). In some embodiments, the VH comprises a HC CDR1 sequence of VSSSSIH (SEQ ID NO: 235). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 240). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 239). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 238). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSSSSIH (SEQ ID NO: 235), a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 236), a HC CDR3 sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 237), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 238), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 239), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 240). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 234). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 233).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 245).
In some embodiments, the VH comprises a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 244). In some embodiments, the VH comprises a HC CDR1 sequence of VSYSSIH (SEQ ID NO: 243). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 248). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 247). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 246). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSYSSIH (SEQ ID NO: 243), a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 244), a HC CDR3 sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 245), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 246), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 247), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 248). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 242). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 241).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYLGMDY (SEQ ID NO: 253).
In some embodiments, the VH comprises a HC CDR2 sequence of YISPYSGYTYYADSVKG (SEQ ID NO: 252). In some embodiments, the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 251). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGSSSLLT (SEQ ID NO: 256). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 255). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 254). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 251), a HC CDR2 sequence of YISPYSGYTYYADSVKG (SEQ ID NO: 252), a HC CDR3 sequence of YWYYLGMDY (SEQ ID NO: 253), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 254), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 255), and a LC CDR3 sequence of QQGSSSLLT (SEQ ID NO: 256). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPYSGY TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYLGMDYWGQG TLVTVSS (SEQ ID NO: 250). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV (SEQ ID NO: 249).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of LWASGLDY; and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VH comprises: (a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (c) a HC CDR3 comprising an amino acid sequence of LWASGLDY. In some embodiments, the VL comprises: (a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of SYYGFWQALWALDY; and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V. In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VH comprises: (a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (c) a HC CDR3 comprising an amino acid sequence of SYYGFWQALWALDY. In some embodiments, the VL comprises: (a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1- 5 amino acid changes; (b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of GYYYPYYAMDY; and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V. In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VH comprises: (a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (c) a HC CDR3 comprising an amino acid sequence of GYYYPYYAMDY. In some embodiments, the VL comprises: (a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of GYYYPYYAMDY; and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V. In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VH comprises: (a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (c) a HC CDR3 comprising an amino acid sequence of GYYYPYYAMDY. In some embodiments, the VL comprises: (a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of X1X2X3X4X5MDY, wherein Xi is F or G, X2 is Q or Y, X3 is W or G, X4 is Y or W, and X5 is A or G; and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V. In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VH comprises: (a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (c) a HC CDR3 comprising an amino acid sequence of X1X2X3X4X5MDY, wherein Xi is F or G, X2 is Q or Y, X3 is W or G, X4 is Y or W, and X5 is A or G. In some embodiments, the VL comprises: (a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of SGYYSSHWYLQSWYQAMDY; and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V. In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VH comprises: (a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (c) a HC CDR3 comprising an amino acid sequence of SGYYSSHWYLQSWYQAMDY. In some embodiments, the VL comprises: (a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of SGYYSSHWYLQSWYQAMDY; and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L. In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VH comprises: (a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (c) a HC CDR3 comprising an amino acid sequence of SGYYSSHWYLQSWYQAMDY. In some embodiments, the VL comprises: (a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of HYSEKWWGWYTMYIDAMDY; and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V. In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VH comprises: (a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (c) a HC CDR3 comprising an amino acid sequence of HYSEKWWGWYTMYIDAMDY. In some embodiments, the VL comprises: (a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1- 5 amino acid changes; (b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L; and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V. In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VH comprises: (a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (c) a HC CDR3 comprising an amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L. In some embodiments, the VL comprises: (a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L; and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L. In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VH comprises: (a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (c) a HC CDR3 comprising an amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L. In some embodiments, the VL comprises: (a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L; and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V. In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VH comprises: (a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (c) a HC CDR3 comprising an amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L. In some embodiments, the VL comprises: (a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of SSRQYYHSQVEPPMAMDY; and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQXIX2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V. In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VH comprises: (a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (c) a HC CDR3 comprising an amino acid sequence of SSRQYYHSQVEPPMAMDY. In some embodiments, the VL comprises: (a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In some embodiments, the antigen-binding domain binds to a peptide conjugate/MHC complex, wherein the peptide conjugate of the peptide conjugate/MHC complex is a peptide covalently linked to a targeted covalent inhibitor or fragment thereof and a MHC. In some embodiments, the targeted covalent inhibitor is adagrasib, sotorasib, or divarasib. In some embodiments, the peptide conjugate/MHC complex is a first peptide conjugate/MHC complex comprising a first peptide conjugate comprising a first peptide and a first targeted covalent inhibitor or fragment thereof; and wherein the antigen-binding domain further binds to a second peptide conjugate/MHC complex comprising (a) a second peptide conjugate, wherein the second peptide conjugate of the second peptide conjugate/MHC complex is a second peptide covalently linked to a second targeted covalent inhibitor or fragment thereof; and (b) a second MHC.
In some embodiments, the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 50 nM and (ii) the second peptide conjugate/MHC complex with a KD of at most about 50 nM. In some embodiments, the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 10 nM and (ii) the second peptide conjugate/MHC complex with a KD of at most about 10 nM. In some embodiments, the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 5 nM and (ii) the second peptide conjugate/MHC complex with a KD of at most about 5 nM.
In some embodiments, (i) the first targeted covalent inhibitor is adagrasib and the second targeted covalent inhibitor is sotorasib, (ii) the first targeted covalent inhibitor is adagrasib and the second targeted covalent inhibitor is divarasib; or (iii) the first targeted covalent inhibitor is divarasib and the second targeted covalent inhibitor is sotorasib. In some embodiments, the antigen-binding domain does not bind to a free first targeted covalent inhibitor with a dissociation constant (KD) of less than 200 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex with a greater affinity than to the first peptide or the free first targeted covalent inhibitor. In some embodiments, the affinity of the antigen-binding domain for the first peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the antigen-binding domain for the first peptide or the free first targeted covalent inhibitor. In some embodiments, the antigen-binding domain does not detectably bind to a complex of the first peptide with a MHC, wherein the first peptide is not covalently bound to the first targeted covalent inhibitor or fragment thereof. In some embodiments, the antigen-binding domain does not detectably bind to the free first targeted covalent inhibitor. In some embodiments, the antigen-binding domain binds to a free first peptide conjugate with a dissociation constant (KD) that is at least 2.5 times more than a KD of the antibody or the antigen-binding fragment to the first peptide conjugate/MHC complex. In some embodiments, the antigen-binding domain does not bind to a free second targeted covalent inhibitor with a dissociation constant (KD) of less than 200 nM.
In some embodiments, the antigen-binding domain binds to the second peptide conjugate/MHC complex with a greater affinity than to the second peptide or the free second targeted covalent inhibitor. In some embodiments, the affinity of the antigen-binding domain for the second peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the antigen-binding domain for the second peptide or the free second targeted covalent inhibitor. In some embodiments, the antigen-binding domain does not detectably bind to a complex of the second peptide with the second MHC, wherein the second peptide is not covalently bound to the second targeted covalent inhibitor or fragment thereof. In some embodiments, the antigen-binding domain does not detectably bind to the free second targeted covalent inhibitor. In some embodiments, the antigen-binding domain binds to a free second peptide conjugate with a dissociation constant KD that is at least 2.5 times more than a KD of the antibody or the antigen-binding fragment to the second peptide conjugate/MHC complex.
In some embodiments, the first peptide and the second peptide comprise the same amino acid sequence. In some embodiments, the first peptide and the second peptide consist of the same amino acid sequence. In some embodiments, the first peptide or the second peptide comprises the amino acid sequence of VVVGACGVGK. In some embodiments, the first peptide and second peptide comprise a different amino acid sequence. In some embodiments, the first peptide or the second peptide comprises an amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV. In some embodiments, the MHC is an HL A, and wherein the HL A i s HL A- A* 02 : 01 , HL A- A* 03 : 01 , or HL A- A* 11 :01.
In some embodiments, the first peptide or the second peptide is or is derived from KRAS. In some embodiments, the first peptide or the second peptide comprises a segment of KRASG12C, KRASG12D, or KRASG12R. In some embodiments, the first peptide conjugate is formed by the covalent reaction of a free first targeted covalent inhibitor with a KRASG12C peptide, a KRASG12D peptide, or a KRASG12R peptide. In some embodiments, the second peptide conjugate is formed by the covalent reaction of a second free targeted covalent inhibitor with a KRASG12C peptide, a KRASG12D peptide, or a KRASG12R peptide. In some embodiments, the free first targeted covalent inhibitor or the second free targeted covalent inhibitor is sotorasib, adagrasib, or divarasib. In some embodiments, the first peptide or the second peptide comprises the amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV.
In some embodiments, the antigen-binding domain binds to (a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01; (b) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01; or (c) a first peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain binds to (a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01; (b) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01; or (c) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain binds to (a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A* 11:01; (b) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01; or (c) a first peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain binds to (a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01; (b) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01; or (c) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain binds to (a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A* 11:01; (b) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01; or (c) a first peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain binds to (a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01; (b) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01; or (c) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain binds to (a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01; (b) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01; or (c) a first peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain binds to (a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01; (b) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01; or (c) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
In some embodiments, the first targeted covalent inhibitor or the second targeted covalent inhibitor is sotorasib, adagrasib, or divarasib.
In some embodiments, the VH is linked to the VL via a linker sequence. In some embodiments, the linker sequence comprises (G4S)n or (S4G)n, wherein n is any integer from 1 to 10. In some embodiments, the linker comprises the glycine-serine-alanine linker G4SA3 or a glycine-serine linker (G4S In some embodiments, the polypeptide is an intact antibody, a bispecific antibody, a multispecific antibody, an antigen-binding (Fab) fragment, an Fab’ fragment, an (Fab’)2 fragment, an Fd, an Fv, a dAb, a single domain fragment or single monomeric variable antibody domain, a single-chain Diabody (scDb), a diabody (Db), a dualaffinity retargeting (DART) molecule, a single-chain variable fragment (scFv), a Bi-specific T-cell engager (BiTE), bispecific killer cell engager (BiKE), CrossMab, a tri-specific binding partner, a chimeric antigen receptor (CAR), a camelid antibody, a monobody (e.g., adnectin), a DARPin, an anticalin, an affibody, or an affimer. In some embodiments, the polypeptide comprises a monobody (e.g., adnectin), a DARPin, an anticalin, an affibody, or an affimer. In some embodiments, the polypeptide comprises a polypeptide chain comprising the antigenbinding domain. In some embodiments, the polypeptide chain further comprises a cytokine or fragment thereof. In some embodiments, cytokine comprises IL-2, IL-7, IL-15, IL-12, IL-18, IL-21, or an interferon (IFN). In some embodiments, the cytokine comprises a modified or mutated IL-2, IL-7, IL-15, IL-12, IL-18, IL-21, or IFN. In some embodiments, the polypeptide chain further comprises an agonist molecule. In some embodiments, the agonist molecule is a CD28 agonist or a 4-1BB agonist.
In an aspect, the present disclosure provides a pharmaceutical composition comprising a polypeptide described herein, and a pharmaceutically acceptable carrier, excipient, adjuvant or diluent.
In an aspect, the present disclosure provides a method of treating cancer in a subject in need thereof wherein the subject has been treated with a free targeted covalent inhibitor, the method comprising administering to the subject the polypeptide described herein or the pharmaceutical composition described herein. In some embodiments, the free targeted covalent inhibitor is sotorasib, adagrasib, or divarasib. In some embodiments, the subject is refractory to a treatment with the free targeted covalent inhibitor.
In an aspect, the present disclosure provides a method of treating a cancer in a subject in need thereof, the method comprising administering to the subject the polypeptide described herein or the pharmaceutical composition described herein. In some embodiments, the method further comprises administering simultaneously a small molecule drug.
In an aspect, the present disclosure provides a method of administering to the subject in need thereof the polypeptide described herein or the pharmaceutical composition described herein, and a free targeted covalent inhibitor, wherein the free targeted covalent inhibitor is optionally sotorasib, adagrasib, or divarasib. In some embodiments, the polypeptide described herein or the pharmaceutical composition described herein is administered after administration of the free targeted covalent inhibitor or simultaneously with the free targeted covalent inhibitor.
In an aspect, the present disclosure provides a method of manufacturing a T-cell receptor (TCR) that recognizes the first peptide conjugate/MHC complex or the second peptide conjugate/MHC complex described herein, the method comprising: (a) a plurality of candidate TCRs with the first peptide conjugate/MHC complex or the second peptide conjugate/MHC complex, and (b) identifying at least one TCR that binds to the first peptide conjugate/MHC complex or the second peptide conjugate/MHC complex. In some embodiments, identifying in (b) further comprises selecting or isolating the at least one TCR.
In some embodiments, the plurality of candidate TCRs is a plurality of soluble TCRs or a plurality of TCRs expressed on cell surface of a plurality of cells. In some embodiments, the plurality of candidate TCRs is the plurality of TCRs expressed on cell surface of the plurality of cells, and identifying in (b) comprises isolating or selecting a cell comprising the at least one TCR based on an activation marker of the cell. In some embodiments, the activation marker is a T cell activation marker. In some embodiments, the T cell activation marker is CD26, CD27, CD28, CD30, CD154, CD40L, CD134. CD25, CD44, CD69, CD 137, PD-1 or KLRGl.
In an aspect, the present disclosure provides a T-cell receptor (TCR) comprising the at least one TCR of (b) of a method described herein. In some embodiments, the TCR is a soluble TCR. In some embodiments, the TCR is a bispecific TCR.
In some embodiments, the first peptide conjugate/MHC complex is different from the second peptide conjugate/MHC complex, wherein each of the first and second peptide conjugate/MHC complexes independently comprises: a different peptide selected from the group consisting of peptides comprising the formula X#X#+iX#+2X#+3X#+4(X#+5)X#+6X#+7X#+sX#+9, and X#X#+ 1 X#+2X#+3 (X#+4)X#+sX#+eX#+7X#+8X#+9, and X#X#+iX#+2(X#+3)X#+4X#+sX#+6X#+7X#+8X#+9, and X#X#+i(X#+2)X#+3X#+4X#+sX#+6X#+7X#+8X#+9, and X#-iX#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9, and X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9, wherein the peptide conjugate of the first and second peptide conjugate/MHC complex is formed by the covalent reaction of the first targeted covalent inhibitor, second targeted covalent inhibitor or fragment thereof with the residue in parenthesis; and (b) the same MHC; wherein the polypeptide binds to the first peptide conjugate/MHC complex with a KD that is at most 100 nM and binds to the second peptide conjugate/MHC complex with an KD that is at most 100 nM, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to a background-subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to the biolayer interferometry analysis, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to a background-subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to the background-subtracted biolayer interferometry analysis, and wherein the reference binding signal is the maximum binding signal of the polypeptide binding to X#X#+ 1 X#+2X#+3X#+4(X#+5)X#+6X#+7X#+8X#+9.
In some embodiments, the formula X#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9 is the formula X7X8X9X10X11X12X13X14X15X16, and wherein X12 is covalently linked to the first targeted covalent inhibitor, second targeted covalent inhibitor or fragment thereof.
In some embodiments, the polypeptide binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex, wherein each of the first and the second peptide conjugate/MHC complex comprises: a peptide comprising the formula X7X8X9X10X11X12X13X14X15X16; and the same MHC; wherein the first peptide conjugate of the first peptide conjugate/MHC complex is formed by the covalent reaction of the first targeted covalent inhibitor or fragment thereof with X12 of the peptide, and wherein the second peptide conjugate of the second peptide conjugate/MHC complex is formed by the covalent reaction of the second targeted covalent inhibitor or fragment thereof with X13 of the peptide, wherein the second targeted covalent inhibitor is the same as the first targeted covalent inhibitor; and wherein the polypeptide binds to the second peptide conjugate/MHC complex with a KD that is at most 100,000-fold higher than the KD of the polypeptide to the first peptide conjugate/MHC complex, or wherein a maximum wavelength shift of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum wavelength shift of the polypeptide binding to the first peptide conjugate/MHC complex, or wherein a maximum binding signal of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum binding signal of the polypeptide binding to the first peptide conjugate/MHC complex.
In some embodiments, the polypeptide binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex, wherein each of the first and the second peptide conjugate/MHC complex comprises: a peptide comprising the formula X7X8X9X10X11X12X13X14X15X16; and the same MHC; wherein the first peptide conjugate of the first peptide conjugate/MHC complex is formed by the covalent reaction of the first targeted covalent inhibitor or fragment thereof with a residue selected from the group consisting of X9, X10, X11, and X12, and wherein the second peptide conjugate of the second peptide conjugate/MHC complex is formed by the covalent reaction of the second targeted covalent inhibitor or fragment thereof with a residue selected from the group consisting of X13, X14, and X15, wherein the second targeted covalent inhibitor is the same as the first targeted covalent inhibitor; wherein the polypeptide binds to the second peptide conjugate/MHC complex with a KD that is at most 100,000 fold higher that the KD of the polypeptide to the first peptide conjugate/MHC complex, or wherein a maximum wavelength shift of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10- fold less than a maximum wavelength shift of the polypeptide binding to the first peptide conjugate/MHC complex, or wherein a maximum binding signal of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum binding signal of the polypeptide binding to the first peptide conjugate/MHC complex.
In some embodiments, the polypeptide binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex, wherein each of the first and the second peptide conjugate/MHC complex comprises: a peptide comprising the formula A-B- C, where A comprises no more than three residues having an N-terminal anchor residue, B comprises at least four but no more than seven residues having a residue covalently linked to the first targeted covalent inhibitor or fragment thereof or the second targeted covalent inhibitor or fragment thereof, and C comprises no more than three residues having a C- terminal anchor residue, wherein the N-terminal anchor residues and the C-terminal anchor residue bind to an MHC; and the same MHC; wherein the residue covalently linked to the first targeted covalent inhibitor or fragment thereof of the peptide of the first peptide conjugate/MHC complex is different from the residue covalently linked to the second targeted covalent inhibitor or fragment thereof of the peptide of the second peptide conjugate/MHC complex; and wherein the polypeptide binds to the first peptide conjugate/MHC complex with a KD that is at most 100 nM and binds to the second peptide conjugate/MHC complex with a KD that is at most 100 nM, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to a background- subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to the background-subtracted biolayer interferometry analysis, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to a background- subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum binding signal of at least 0.1 -fold of a reference binding signal is observed according to the background- subtracted biolayer interferometry analysis, and wherein the reference binding signal is the maximum binding signal of the polypeptide binding to X#X#+ 1 X#+2X#+3X#+4(X#+5)X#+6X#+7X#+8X#+9. In some embodiments, A comprises two residues X?Xs. In some embodiments, B comprises seven residues X9X10X11X12X13X14X15. In some embodiments, C comprises one residue Xi6.
In some embodiments, the formula A-B-C is the formula X7X8X9X10X11X12X13X14X15X16. In some embodiments, the first peptide conjugate of the first peptide conjugate/MHC complex is formed by the covalent reaction of the first targeted covalent inhibitor or fragment thereof with X12 of the peptide. In some embodiments, the second peptide conjugate of the second peptide conjugate/MHC complex is formed by the covalent reaction of the second targeted covalent inhibitor or fragment thereof with X13 of the peptide, wherein the second targeted covalent inhibitor is the same as the first targeted covalent inhibitor. In some embodiments, the polypeptide binds to the second peptide conjugate/MHC complex with a KD that is at most 100,000 fold higher than the KD of the polypeptide to the first peptide conjugate/MHC complex.
In some embodiments, the polypeptide binds to the second peptide conjugate/MHC complex with an KD that is at most 100,000 fold higher than the KD of the polypeptide to the first peptide conjugate/MHC complex, and wherein the residue covalently linked to the first targeted covalent inhibitor or fragment thereof of the peptide of the first peptide conjugate/MHC complex is selected from the group of consisting of X9, X10, X11, and X12, and the residue covalently linked to the second targeted covalent inhibitor or fragment thereof of the peptide of the second peptide conjugate/MHC complex is X13, X14, and X15.
In some embodiments, the peptide is a RAS peptide. In some embodiments, the RAS peptide comprises a mutation. In some embodiments, the mutation is G12C or G13C. In some embodiments, the RAS peptide comprises a sequence selected from the group consisting of VVVGACGVGK, VVGACGVGK, and KLVVVGACGV. In some embodiments, the RAS peptide comprises a sequence selected from the group consisting of VVVGAGCVGK, VVGAGCVGK, or KLVVVGAGCV. In some embodiments, the same MHC is selected from the group consisting of HL A-A* 03:01, HLA-A* 11 :01, HLA-A*02:01, HLA-A*68:01, HLA- A*31:01, HLA-A*30:01, HLA-A*33:03, HLA-A*33:01, HLA-A*74:01, HLA-A*34:02, HLA-A*66:01, HLA-A*68:02, HLA-A*02:05, HLA-A*02:02, and HLA-A* 02: 06. In some embodiments, the targeted covalent inhibitor or fragment thereof comprises sotorasib. In some embodiments, the peptide is a RAS peptide, and wherein X12 is G12C mutation, and X13 is G13C mutation.
In some embodiments, the polypeptide binds to the first peptide conjugate/MHC complex having a Cys of the G12C mutation covalently linked to the first targeted covalent inhibitor or fragment thereof with a KD that is at most 100 nM and binds to the second peptide conjugate/MHC complex having a Cys of the G13C mutation covalently linked to the second targeted covalent inhibitor or fragment thereof with an KD that is most 100 nM, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to a background- subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to the background- subtracted biolayer interferometry analysis, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to a background-subtracted biolayer interferometry analysis and wherein when a polypeptide is contact to the second peptide conjugate/MHC complex a maximum binding signal of at least 0.1 -fold of a reference binding signal is observed according to a background- subtracted biolayer interferometry analysis, and wherein the reference binding signal is the maximum binding signal of the polypeptide binding to X#X#+lX#+2X#+3X#+4X#+5X#+6X#+7X#+8X#+9.
In some embodiments, the polypeptide binds to the second peptide conjugate/MHC complex having a Cys of the G13C mutation covalently linked to the second targeted covalent inhibitor or fragment thereof with a KD that is at most 100,000 fold higher that the KD of the polypeptide to the first peptide conjugate/MHC complex having a Cys of the G12C mutation covalently linked to the first targeted covalent inhibitor or fragment thereof, or wherein a maximum wavelength shift of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum wavelength shift of the polypeptide binding to the first peptide conjugate/MHC complex, or wherein a maximum binding signal of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum binding signal of the polypeptide binding to the first peptide conjugate/MHC complex.
In some aspects, the present disclosure provides a composition comprising: a polypeptide that binds to a first peptide conjugate/MHC complex and a second peptide conjugate/MHC complex different from the first peptide conjugate/MHC complex, wherein each of the first and second peptide conjugate/MHC complexes independently comprises: a different peptide selected from the group consisting of peptides comprising the formula X#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9, and X#X#+ 1 X#+2X#+3 (X#+4)X#+sX#+eX#+7X#+8X#+9, and X#X#+iX#+2(X#+3)X#+4X#+sX#+6X#+7X#+8X#+9, and X#X#+i(X#+2)X#+3X#+4X#+sX#+6X#+7X#+8X#+9, and X#-iX#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9, and X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9, wherein the peptide conjugate of the first and second peptide conjugate/MHC complex is formed by the covalent reaction of a targeted covalent inhibitor or fragment thereof with the residue in parenthesis; and (b) the same MHC; wherein the polypeptide binds to the first peptide conjugate/MHC complex with an KD that is at most 100 nM and binds to the second peptide conjugate/MHC complex with an KD that is at most 100 nM, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to a background-subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to the background- subtracted biolayer interferometry analysis, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum binding signal of at least 0.1 -fold of a reference binding signal is observed according to a background-subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to the background-subtracted biolayer interferometry analysis, and wherein the reference binding signal is the maximum binding signal of the polypeptide binding to X#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9.
In some embodiments, each of the first and second peptide conjugate/MHC complexes independently comprises: a different peptide selected from the group consisting of peptides comprising the formula X#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9, and X#X#+ 1 X#+2X#+3 (X#+4)X#+sX#+eX#+7X#+8X#+9, and X#X#+iX#+2(X#+3)X#+4X#+sX#+6X#+7X#+8X#+9, and X#X#+l(X#+2)X#+3X#+4X#+5X#+6X#+7X#+8X#+9.
In some embodiments, each of the first and second peptide conjugate/MHC complexes independently comprises: a different peptide with an amino acid sequence selected from the group consisting of: VVVGACGVGK, VVVGCGGVGK, VVVCAGGVGK, and VVCGAGGVGK.
In some aspects, the present disclosure provides a composition that binds to a first peptide conjugate/MHC complex and a second peptide conjugate/MHC complex, wherein each of the first and the second peptide conjugate/MHC complex comprises: a peptide comprising the formula X7X8X9X10X11X12X13X14X15X16; and the same MHC; wherein a first peptide conjugate of the first peptide conjugate/MHC complex is formed by the covalent reaction of a targeted covalent inhibitor or fragment thereof with X12 of the peptide, and wherein a second peptide conjugate of the second peptide conjugate/MHC complex is formed by the covalent reaction of the same targeted covalent inhibitor or fragment thereof with X13 of the peptide; wherein a polypeptide binds to the second peptide conjugate/MHC complex with a KD that is at most 100,000 fold higher than the KD of the polypeptide to the first peptide conjugate/MHC complex, or wherein a maximum wavelength shift of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum wavelength shift of the polypeptide binding to the first peptide conjugate/MHC complex, or wherein a maximum binding signal of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum binding signal of the polypeptide binding to the first peptide conjugate/MHC complex.
In some aspects, the present disclosure provides a composition that binds to a first peptide conjugate/MHC complex and a second peptide conjugate/MHC complex, wherein each of the first and the second peptide conjugate/MHC complex comprises: a peptide comprising the formula X7X8X9X10X11X12X13X14X15X16; and the same MHC; wherein a first peptide conjugate of the first peptide conjugate/MHC complex is formed by the covalent reaction of a targeted covalent inhibitor or fragment thereof with a residue selected from the group consisting of X9, X10, X11, and X12, and wherein a second peptide conjugate of the second peptide conjugate/MHC complex is formed by the covalent reaction of the same targeted covalent inhibitor or fragment thereof with a residue selected from the group consisting of X13, X14, and X15; wherein a polypeptide binds to the second peptide conjugate/MHC com-plex with a KD that is at most 100,000 fold higher that the KD of the polypeptide to the first peptide conjugate/MHC complex, or wherein a maximum wavelength shift of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum wavelength shift of the polypeptide binding to the first peptide conjugate/MHC complex, or wherein a maximum binding signal of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum binding signal of the polypeptide binding to the first peptide conjugate/MHC complex.
In some aspects, the present disclosure provides a composition that binds to a first peptide conjugate/MHC complex and a second peptide conjugate/MHC complex, wherein each of the first and the second peptide conjugate/MHC complex comprises: a peptide comprising the formula A-B-C, where A comprises no more than three residues having an N- terminal anchor residue, B comprises at least four but no more than seven residues having a residue covalently linked to a targeted covalent inhibitor or fragment thereof, and C comprises no more than three residues having a C-terminal anchor residue, wherein the N- terminal anchor residues and the C-terminal anchor residue bind to an MHC; and the same MHC; wherein the residue covalently linked to a targeted covalent inhibitor or fragment thereof of the peptide of the first peptide conjugate/MHC complex is different from the residue covalently linked to a targeted covalent inhibitor or fragment thereof of the peptide of the second peptide conjugate/MHC complex; and wherein a polypeptide binds to the first peptide conjugate/MHC complex with a KD that is at most 100 nM and binds to the second peptide conjugate/MHC complex with a KD that is at most 100 nM, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to a background- subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to the background-subtracted biolayer interferometry analysis, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum binding signal of at least 0.1 -fold of a reference binding signal is observed according to a background- subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum binding signal of at least 0.1 -fold of a reference binding signal is observed according to the background- subtracted biolayer interferometry analysis, and wherein the reference binding signal is the maximum binding signal of the polypeptide binding to X#X#+ 1 X#+2X#+3X#+4(X#+5)X#+6X#+7X#+8X#+9.
In some embodiments, A comprises two residues X?Xs. In some embodiments, B comprises seven residues X9X10X11X12X13X14X15. In some embodiments, C comprises one residue Xi6. In some embodiments, the formula A-B-C is the formula X7X8X9X10X11X12X13X14X15X16. In some embodiments, a first peptide conjugate of the first peptide conjugate/MHC complex is formed by the covalent reaction of the targeted covalent inhibitor or fragment thereof with X12 of the peptide. In some embodiments, a second peptide conjugate of the second peptide conjugate/MHC complex is formed by the covalent reaction of the same targeted covalent inhibitor or fragment thereof with X13 of the peptide. In some embodiments, the polypeptide binds to the second peptide conjugate/MHC complex with a KD that is at most 100,000 fold higher than the KD of the polypeptide to the first peptide conjugate/MHC complex.
In some embodiments, the polypeptide binds to the second peptide conjugate/MHC complex with an KD that is at most 100,000 fold higher than the KD of the polypeptide to the first peptide conjugate/MHC complex, and wherein the residue covalently linked to the targeted covalent inhibitor or fragment thereof of the peptide of the first peptide conjugate/MHC complex is selected from the group of consisting of X9, X10, X11, and X12, and the residue covalently linked to the targeted covalent inhibitor or fragment thereof of the peptide of the second peptide conjugate/MHC complex is X13, X14, and X15.
In some embodiments, the peptide is a RAS peptide. In some embodiments, the RAS peptide comprises a mutation. In some embodiments, the mutation is G12C or G13C. In some embodiments, the RAS peptide comprises a sequence selected from the group consisting of VVVGACGVGK, VVGACGVGK, and KLVVVGACGV. In some embodiments, the RAS peptide comprises a sequence selected from the group consisting of VVVGAGCVGK, VVGAGCVGK, or KLVVVGAGCV. In some embodiments, the same MHC is selected from the group consisting of HL A- A* 03: 01, HLA-A* 11:01, HLA-A*02:01, HLA-, HLA- A*68:01, HLA-A*31:01, HLA-A*30:01, HLA-A*33:03, HLA-A*33:01, HLA-A*74:01, HLA-A*34:02, HLA-A*66:01, HLA-A*68:02, HLA-A*02:05, HLA-A*02:02, and HLA- A*02:06. In some embodiments, the targeted covalent inhibitor or fragment thereof comprises sotorasib. In some embodiments, the peptide is a RAS peptide, and wherein X12 is G12C mutation, and X13 is G13C mutation.
In some embodiments, the polypeptide binds to the first peptide conjugate/MHC complex having a Cys of the G12C mutation covalently linked to the targeted covalent inhibitor or fragment thereof with an KD that is at most 100 nM and binds to the second peptide conjugate/MHC complex having a Cys of the G13C mutation covalently linked to the targeted covalent inhibitor or fragment thereof with an KD that is at most 100 nM, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to a background- subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to the background-subtracted biolayer interferometry analysis, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to a background- subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum binding signal of at least 0.1 -fold of a reference binding signal is observed according to the background- subtracted biolayer interferometry analysis, and wherein the reference binding signal is the maximum binding signal of the polypeptide binding to X#X#+ 1 X#+2X#+3X#+4(X#+5)X#+6X#+7X#+8X#^9.
In some embodiments, the polypeptide binds to the second peptide conjugate/MHC complex having a Cys of the G13C mutation covalently linked to the targeted covalent inhibitor or fragment thereof with a KD that is at most 100,000 fold higher that the KD of the polypeptide to the first peptide conjugate/MHC complex having a Cys of the G12C mutation covalently linked to the targeted covalent inhibitor or fragment thereof, or wherein a maximum wavelength shift of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum wavelength shift of the polypeptide binding to the first peptide conjugate/MHC complex, or wherein a maximum binding signal of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum binding signal of the polypeptide binding to the first peptide conjugate/MHC complex.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein: (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of FX2X3X4AMDY, wherein X2 is Y, L, T, H, E, A, or R; X3 is D, V, L, E, H, T or R; and X4 is L or Y (SEQ ID NO: 267); and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX3SWLX7WX9X10T, wherein X3 is A or S, X7 is Y or H, X9 is L, K, V, Y or I, and X10 is L, V, or I (SEQ ID NO: 268).
In some embodiments, the VH comprises: a HC CDR3 comprising the amino acid sequence of FX2X3X4AMDY, wherein X2 is Q, Y, L, T, H, E, A, or R; X3 is W, D, V, L, E, H, T or R; and X4 is L or Y.
In some embodiments, the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-3 amino acid changes. In some embodiments, the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of FX2X3X4AMDY, wherein X2 is Y, L, T, H, E, A, or R; X3 is D, V, L, E, H, T or R; and X4 is L or Y. In some embodiments, the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-3 amino acid changes, and/or a LC CDR3 comprising the amino acid sequence of QQX3SWLX7WX9X10T, wherein X3 is A or S, X7 is Y or H, X9 is L, K, V, Y or I, and X10 is L, V, or I.
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FYDLAMDY (SEQ ID NO.: 273).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 272). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 271). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWLIT (SEQ ID NO.: 276). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 275). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 274). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 271), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 272), a HC CDR3 sequence of FYDLAMDY (SEQ ID NO: 273), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 274), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 275), and a LC CDR3 sequence of QQASWLYWLIT (SEQ ID NO: 276). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMDYWGQGTLV TVSS (SEQ ID NO.: 270). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 269).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FLDLAMDY (SEQ ID NO.: 281). In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 280). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 279). In some embodiments, the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWKIT (SEQ ID NO.: 284). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 283). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 282). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 279), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 280), a HC CDR3 sequence of FLDLAMDY (SEQ ID NO: 281), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 282), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 283), and a LC CDR3 sequence of QQASWLYWKIT (SEQ ID NO: 284). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFLDLAMDYWGQGTLV TVSS (SEQ ID NO.: 278). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKITFGQGTKVEIK (SEQ ID NO.: 277).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTVLAMDY (SEQ ID NO.: 289).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 288). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 287). In some embodiments, the antigen- binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWLIT (SEQ ID NO.: 292). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 291). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 290). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 287), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 288), a HC CDR3 sequence of FTVLAMDY (SEQ ID NO.: 289), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 290), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 291), and a LC CDR3 sequence of QQASWLYWLIT (SEQ ID NO.: 292). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 286). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 285).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTVLAMDY (SEQ ID NO.: 297).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 296). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 295). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWKVT (SEQ ID NO.: 300). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 299). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 298). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 295), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 296), a HC CDR3 sequence of FTVLAMDY (SEQ ID NO.: 297), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 297), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 298), and a LC CDR3 sequence of QQASWLYWKVT (SEQ ID NO.: 300). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSS WVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 294). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKVTFGQGTKVEIK (SEQ ID NO.: 293).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTVLAMDY (SEQ ID NO.: 305).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 304). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 303). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWVIT (SEQ ID NO.: 308). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 307). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 306). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 303), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 304), a HC CDR3 sequence of FTVLAMDY (SEQ ID NO.: 305), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 306), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 307), and a LC CDR3 sequence of QQASWLYWVIT (SEQ ID NO.: 308). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 302). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWVITFGQGTKVEIK (SEQ ID NO.: 301). In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FHDLAMDY (SEQ ID NO.: 313).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 312). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 311). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWKIT (SEQ ID NO.: 316). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 315). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 314). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 311), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 312), a HC CDR3 sequence of FHDLAMDY (SEQ ID NO.: 313), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 314), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 315), and a LC CDR3 sequence of QQASWLYWKIT (SEQ ID NO.: 316). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHDLAMDYWGQGTLV TVSS (SEQ ID NO.: 310). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKITFGQGTKVEIK (SEQ ID NO.: 309).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FHELAMDY (SEQ ID NO.: 321).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 320). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 319). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWLIT (SEQ ID NO.: 324). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 323). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 322). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 319), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 320), a HC CDR3 sequence of FHELAMDY (SEQ ID NO.: 321), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 322), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 323), and a LC CDR3 sequence of QQASWLYWLIT (SEQ ID NO.: 324). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMDYWGQGTLV TVSS (SEQ ID NO.: 318). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 317).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTVLAMDY (SEQ ID NO.: 329).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 328). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 327). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWKIT (SEQ ID NO.: 332). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 331). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 330). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 327), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 328), a HC CDR3 sequence of FTVLAMDY (SEQ ID NO.: 329), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 330), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 331), and a LC CDR3 sequence of QQSSWLYWKIT (SEQ ID NO.: 332). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 326). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKVEIK (SEQ ID NO.: 325).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FELLAMDY (SEQ ID NO.: 337).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 336). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 335). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWKIT (SEQ ID NO.: 340). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 339). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 338). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 335), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 336), a HC CDR3 sequence of FELLAMDY (SEQ ID NO.: 337), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 338), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 339), and a LC CDR3 sequence of QQSSWLYWKIT (SEQ ID NO.: 340). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFELLAMDYWGQGTLV TVSS (SEQ ID NO.: 334). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKVEIK (SEQ ID NO.: 333).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTVLAMDY (SEQ ID NO.: 345).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 344). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 343). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWKVT (SEQ ID NO.: 348). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 347). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 346). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 343), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 344), a HC CDR3 sequence of FTVLAMDY (SEQ ID NO.: 345), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 346), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 347), and a LC CDR3 sequence of QQSSWLYWKVT (SEQ ID NO.: 348). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 342). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKVTFGQGTKVEIK (SEQ ID NO.: 341).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FYDLAMDY (SEQ ID NO.: 353).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 352). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 351). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWKVT (SEQ ID NO.: 356). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 355). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 354). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 351), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 352), a HC CDR3 sequence of FYDLAMDY (SEQ ID NO.: 353), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 354), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 355), and a LC CDR3 sequence of QQSSWLYWKVT (SEQ ID NO.: 356). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMDYWGQGTLV TVSS (SEQ ID NO.: 350). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKVTFGQGTKVEIK (SEQ ID NO.: 349).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FHELAMDY (SEQ ID NO.: 361).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 360). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 359). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWLIT (SEQ ID NO.: 364). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 363). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 362). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 359), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 360), a HC CDR3 sequence of FHELAMDY (SEQ ID NO.: 361), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 362), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 363), and a LC CDR3 sequence of QQASWLYWLIT (SEQ ID NO.: 364). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMDYWGQGTLV TVSS (SEQ ID NO.: 358). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 357).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTVLAMDY (SEQ ID NO.: 369).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 368). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 367). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWKVT (SEQ ID NO.: 372). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 371). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 370). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 367), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 368), a HC CDR3 sequence of FTVLAMDY (SEQ ID NO.: 369), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 370), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 371), and a LC CDR3 sequence of QQASWLYWKVT (SEQ ID NO.: 372). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 366). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKVTFGQGTKVEIK (SEQ ID NO.: 365).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FYDLAMDY (SEQ ID NO.: 377). In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 376). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 375). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWVIT (SEQ ID NO.: 380). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 379). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 378). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 375), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 376), a HC CDR3 sequence of FYDLAMDY (SEQ ID NO.: 377), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 378), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 379), and a LC CDR3 sequence of QQSSWLYWVIT (SEQ ID NO.: 380). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMDYWGQGTLV TVSS (SEQ ID NO.: 374). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWVITFGQGTKVEIK (SEQ ID NO.: 373).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FHELAMDY (SEQ ID NO.: 385).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 384). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 383). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWLIT (SEQ ID NO.: 388). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 387). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 386). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 383), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 384), a HC CDR3 sequence of FHELAMDY (SEQ ID NO.: 385), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 386), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 387), and a LC CDR3 sequence of QQASWLYWLIT (SEQ ID NO.: 388). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMDYWGQGTLV TVSS (SEQ ID NO.: 382). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 381).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FAHLAMDY (SEQ ID NO.: 393).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 392). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 391). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWLIT (SEQ ID NO.: 396). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 395). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 394). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 391), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 392), a HC CDR3 sequence of FAHLAMDY (SEQ ID NO.: 393), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 394), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 395), and a LC CDR3 sequence of QQSSWLYWLIT (SEQ ID NO.: 396). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFAHLAMDYWGQGTLV TVSS (SEQ ID NO.: 390). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWLITFGQGTKVEIK (SEQ ID NO.: 389).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FEDLAMDY (SEQ ID NO.: 401).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 400). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 399). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWYIT (SEQ ID NO.: 404). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 403). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 402). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 399), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 400), a HC CDR3 sequence of FEDLAMDY (SEQ ID NO.: 401), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 402), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 403), and a LC CDR3 sequence of QQASWLYWYIT (SEQ ID NO.: 404). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFEDLAMDYWGQGTLV TVSS (SEQ ID NO.: 398). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWYITFGQGTKVEIK (SEQ ID NO.: 397).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FRTLAMDY (SEQ ID NO.: 409).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 408). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 407). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWIIT (SEQ ID NO.: 412). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 411). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 410). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 407), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 408), a HC CDR3 sequence of FRTLAMDY (SEQ ID NO.: 409), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 410), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 411), and a LC CDR3 sequence of QQSSWLYWIIT (SEQ ID NO.: 412). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFRTLAMDYWGQGTLV TVSS (SEQ ID NO.: 406). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWIITFGQGTKVEIK (SEQ ID NO.: 405).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTHYAMDY (SEQ ID NO.: 417).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 416). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 415). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWKIT (SEQ ID NO.: 420). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 419). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 418). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 415), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 416), a HC CDR3 sequence of FTHYAMDY (SEQ ID NO.: 417), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 418), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 419), and a LC CDR3 sequence of QQSSWLYWKIT (SEQ ID NO.: 420). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSS WVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTHYAMDYWGQGTLV TVSS (SEQ ID NO.: 414). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKVEIK (SEQ ID NO.: 413).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FRLYAMDY (SEQ ID NO.: 425).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 424). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 423). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLHWKLT (SEQ ID NO.: 428). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 427). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 426). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 423), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 424), a HC CDR3 sequence of FRLYAMDY (SEQ ID NO.: 425), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 426), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 427), and a LC CDR3 sequence of QQASWLHWKLT (SEQ ID NO.: 428). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFRLYAMDYWGQGTLV TVSS (SEQ ID NO.: 422). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLHWKLTFGQGTKVEIK (SEQ ID NO.: 421).
In an aspect, the present disclosure provides a composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FHRLAMDY (SEQ ID NO.: 433).
In some embodiments, the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 432). In some embodiments, the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 431). In some embodiments, the antigenbinding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLHWKLT (SEQ ID NO.: 436). In some embodiments, the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 435). In some embodiments, the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 434). In some embodiments, the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 431), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 432), a HC CDR3 sequence of FHRLAMDY (SEQ ID NO.: 433), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 434), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 435), and a LC CDR3 sequence of QQASWLHWKLT (SEQ ID NO.: 436). In some embodiments, the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHRLAMDYWGQGTLV TVSS (SEQ ID NO.: 430). In some embodiments, the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLHWKLTFGQGTKVEIK (SEQ ID NO.: 429).
In an example, a method of the disclosure includes a method of treating an individual who has received a drug that forms a peptide-drug conjugate by administering a means for specifically binding the peptide-drug conjugate in a complex with an MCH. The means may bind with greater affinity to the peptide-drug conjugate in a complex with an MCH than the affinity the means has for the free drug, or may not detectably bind the free drug. The method of using the means may be used with an individual in need of the means, including but not necessarily limited to a cancer patient, who may have a type of cancer that is resistant to the drug. In an example, the means may bind to two different peptide-drug conjugates that are each present in a different MHC complex.
BRIEF DESCRIPTION OF THE FIGURES
FIG. 1 depicts binding of antibody clones to peptide conjugate/MHC complex. The antibody clones were displayed on the yeast cell surface, and binding of the targets conjugated to fluorescently labeled streptavidin was detected by using flow cytometry.
FIGs. 2A-2C depict binding properties of the RM 010 Fab to peptide conjugate/MHC complex. FIG. 2A shows biolayer interferometry (BLI) sensorgrams of the interaction between peptide conjugate/MHC complexes, with either sotorasib or adagrasib and either HLA-A*03 or HLA-A*11. KD values were estimated from a global fitting curve. FIG. 2B shows BLI sensorgrams of the interaction between RM 010 Fab and adagrasib- P7/HLA-A*11 in the presence or absence of free adagrasib drug. FIG. 2C shows plotted MHC binding as a function of free-adagrasib concentration. Binding signal intensity was normalized from the value with no free adagrasib and the 0 nm baseline.
FIGs. 3A-3C depict results of a sotorasib scan. FIG. 3A shows a schematic illustration of the method and the scheme for residues in the panel. Anchor residues are underlined in the sequence. FIG. 3B shows BLI sensorgrams of the binder to the tested soto- p?/MHC complex immobilized on sensor tips. FIG. 3C shows binding signal of the Fab binder to the tested soto-p7/MHC complexes at 1200 s of the measurement (e.g., Bmax, indicated as the vertical line in FIG. 3B).
FIGs. 4A-4C depict results of a HLA scan. FIG. 4A shows schematic illustrations of the modified soto-p7 peptides used in the HLA scan. Anchor residues are underlined and the sotorasib -conjugated Cys is highlighted. FIG. 4B shows BLI sensorgrams of the binder to the tested soto-p7/MHC complexes. FIG. 4C shows binding signal of the Fab binder to the tested soto-p7/MHC complexes at 1200 s of the measurement (e.g., Bmax, indicated as the vertical line in FIG. 4B).
FIGs. 5A-5C depict results of a deep mutational scanning and analysis of the variable light (VL) chain region for binding partner RM 010. FIG. 5A shows the residues (underlined) that were diversified in the mutational scanning. FIG. 5B shows enrichment of variants across tested VL positions for adagrasib-p7 in complex with HLA-A*03. FIG. 5C shows enrichment of variants across tested VL positions for adagrasib-p7 in complex with HLA-A*11. FIGs. 6A-6C depict results of a deep mutational scanning and analysis of the variable heavy (VH) chain region for binding partner RM 010. FIG. 6A shows the residues (underlined) that were diversified in the mutational scanning. FIG. 6B shows enrichment of variants across tested VH positions for adagrasib-p7 in complex with HLA-A*03. FIG. 6C shows enrichment of variants across tested VH positions for adagrasib-p7 in complex with HLA-A*11.
FIG. 7 depicts results of a drug cross-reactivity scan to assess binding response of binding partner RM 010 for peptide-conjugate/MHC complexes with adagrasib (ada- p7/Al l), sotorasib (soto-p7/Al l), or divarasib (GDC6036-p7/Al l). RM_010 showed binding to all complexes at different affinities.
FIG. 8 depicts results of a sotorasib scan. The location of the covalently bound targeted covalent inhibitor (e.g., sotorasib) on the peptide was varied by introducing a unique cysteine residue to replace the residue at position 7, 9, 10, 11, 12, 13, 14, or 15 of peptide VVVGAGGVGK. Binding partner RM 010 showed the strongest binding response to the peptide-conjugate/MHC complex with haptenated p7 peptide at position 12 (e.g., cysteine residue at position 12 conjugated to the targeted covalent inhibitor).
FIG. 9 depicts binding of antibody clones to peptide conjugate/MHC complex with adagrasib (adagrasib-p7/Al 1). Multiple binders showed higher MFI signals after 60 minutes of dissociation in the presence of a nonbiotinylated competitor, compared to MFI signals of the parent clone RM010.
FIG. 10 depicts binding of antibody clones to peptide conjugate/MHC complex with adagrasib (ada-p7/Al 1 or ada-p7/A03). Multiple binders showed higher MFI signals for ada- p7/Al 1 compared to parent clone RM010, indicating stronger binding. Binder RM119 showed higher MFI signal for ada-p7/03 compared to parent clone RM010, indicating stronger binding for this peptide conjugate/MHC complex.
DETAILED DESCRIPTION
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
Every numerical range given throughout this specification includes its upper and lower values, as well as every narrower numerical range that falls within it, as if such narrower numerical ranges were all expressly written herein. As used in the specification and the appended claims, the singular forms “a” "and” and “the" include plural referents unless the context clearly dictates otherwise. Ranges may be expressed herein as from “about” one particular value, and/or to “about” another particular value. When such a range is expressed, another embodiment includes from the one particular value and/or to the other particular value. Similarly, when values are expressed as approximations, by the use of the antecedent “about” it will be understood that the particular value forms another embodiment. The term “about” in relation to a numerical value encompasses variations of +/-10%, +/- 5%, or +/- 1%.
This disclosure includes every amino acid sequence described herein and all nucleotide sequences encoding the amino acid sequences. Every antibody sequence and antigen-binding fragments of them are included. Polynucleotide and amino acid sequences having from 80-99% similarity, inclusive, and including all numbers and ranges of numbers there between, with the sequences provided herein, are included in the invention. All of the amino acid sequences described herein can include amino acid substitutions, such as conservative substitutions, that do not adversely affect the function of the protein that comprises the amino acid sequences. In this regard, the disclosure provides alternative residues for certain positions in described binding partner as described below. In certain examples, the alternative residues were identified by deep mutational scanning, which demonstrates binding functionality for each binding partner that contains the described amino acid change(s). The disclosure includes each binding partner with each alternative residue substituted for the original residue alone and in any combination with the described alternative residues. Thus, any binding partner described herein may have any single described residue change or a combination of described changes. Representative changes for particular antibodies are described the Tables. The changes may be in CDR1, CDR2, CDR3, and combinations thereof. The changes can also include amino acid insertions. The disclosure includes each amino acid sequence that is encompassed by the description of alternative amino acids by reference to a specific sequence identifier and those described in the aforementioned Tables.
As described above, the present disclosure provides antibodies and antigen-binding domains or fragments thereof (collectively “binding partners” and each individually a “binding partner”). The term “antibody” includes each binding partner format herein. The antibody can comprise a polypeptide with an antigen-binding domain or fragment thereof. The binding partners bind with specificity to a protein or fragment thereof, or a peptide provided in peptide form, that comprises a covalently attached molecule. The covalently attached molecule forms a peptide conjugate. A “peptide conjugate” as used herein means any protein or peptide that has been modified so that it is covalently conjugated to another molecule. The peptide conjugate is considered to be a novel antigen, i.e., a neoantigen. The other molecule that is covalently conjugated to the protein or peptide to form the peptide conjugate is not particularly limited, with the proviso that the other molecule is not an additional amino acid that is added to the described peptide conjugates. In embodiments, the molecule that is covalently conjugated to the protein or peptide has or had biological activity before conjugation, or it may be biologically inert before conjugation. In embodiments, the molecule is a drug, including but not necessarily limited to small molecule drugs. As used herein, the molecule that is covalently attached to a peptide to form peptide conjugate is referred to as a “targeted covalent inhibitor (TCI)” or as a “covalent drug.” Representative and non-limiting examples of drugs that covalently attach to a peptide or protein to form a peptide conjugate are described below. Peptide conjugates include but are not limited to covalently modified full length proteins and fragments thereof. Peptide conjugates include fragments of full length proteins that include a covalent modification and are produced, for example, by intracellular processing. In certain embodiments, a full length protein may be covalently modified within a cell and subsequently processed such that a peptide conjugate that is a fragment of the full length protein is produced. In an embodiment, the peptide conjugate comprises a fragment of a full-length protein. As described further below, the produced peptide conjugate may be displayed on a cell surface. The cell surface display of the peptide conjugate may be any form of cell surface display, including but not limited to by way of any receptor having an extracellular segment, or it may be displayed by way of any type of major histocompatibility complex (MHC) or human leukocyte antigen (HLA). Nonlimiting examples of HLA types that display peptide conjugates, and to which the described binding partners bind with specificity, are described further below.
As used herein, the term “peptide conjugate/MHC complex” refers to a peptide conjugate comprising: a peptide and a chemical fragment of a targeted covalent inhibitor, presented by a major histocompatibility complex (MHC). For example, the peptide conjugate can be formed by the covalent reaction of a targeted covalent inhibitor with a residue (e.g., a cysteine residue) in a peptide. In some embodiments, the peptide conjugate is formed by the covalent reaction of AMG-510 with a KRASG12C peptide. In some embodiments, the peptide is externally introduced as a vaccine. In some embodiments, the peptide comprises a nucleophilic or an electrophilic residue. In some embodiments, the residue comprises cysteine, aspartic acid, or arginine. In an embodiment, the MHC is a human leukocyte antigen (HLA). In an embodiment, the HLA is HLA-A*02:01, HLA-A*03:01, or HLA-A*11 :01.
As used herein, the term “CDR” or “complementarity determining region” means the noncontiguous antigen combining sites found within the variable regions of heavy and light chain polypeptides. These particular regions have been described by, for example, Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest (1991), by Chothia et al., J. Mol. Biol. 196:901-917 (1987), and by MacCallum et al., J. Mol. Biol. 262:732-745 (1996), all of which are herein incorporated by reference in their entireties, where the definitions include overlapping or subsets of amino acid residues when compared against each other. In certain embodiments, the term “CDR” is a CDR as defined by MacCallum et al., J. Mol. Biol. 262:732-745 (1996) and Martin A. “Protein Sequence and Structure Analysis of Antibody Variable Domains,” in Antibody Engineering, Kontermann and Diibel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001). In certain embodiments, the term “CDR” is a CDR as defined by Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest (1991). In certain embodiments, heavy chain CDRs and light chain CDRs of an antibody are defined using different conventions. In certain embodiments, heavy chain CDRs and/or light chain CDRs are defined by performing structural analysis of an antibody and identifying residues in the variable region(s) predicted to make contact with an epitope region of a target molecule (e.g., a peptide conjugate). HC CDR1, HC CDR2, and HC CDR3 denote the heavy chain CDRs, and LC CDR1, LC CDR2 and LC CDR3 denote the light chain CDRs. The CDRs for any binders (e.g., binding partners or antigen binding domains) described herein can be designated by Kabat numbering scheme. In some cases, the light chain (LC) CDRs can be designated by Kabat numbering scheme. In some cases, the LC CDRs can be designated by Kabat numbering scheme with modifications. In some cases, the heavy chain (HC) CDRs can be designated by Kabat numbering scheme. In some cases, the HC CDRs can be designated by Kabat numbering scheme with modifications. For example, the CDRs may contain one or more extra amino acids than the CDRs designated by Kabat numbering scheme.
The determination of “percent identity” between two sequences (e.g., amino acid sequences or nucleic acid sequences) can be accomplished using a mathematical algorithm. A specific, non-limiting example of a mathematical algorithm utilized for the comparison of two sequences is the algorithm of Karlin S & Altschul SF (1990) PNAS 87: 2264-2268, modified as in Karlin S & Altschul SF (1993) PNAS 90: 5873-5877, each of which is herein incorporated by reference in its entirety. Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul SF et al., (1990) J Mol Biol 215: 403, which is herein incorporated by reference in its entirety. BLAST nucleotide searches can be performed with the NBLAST nucleotide program parameters set, e.g., for score=100, wordlength=12 to obtain nucleotide sequences homologous to a nucleic acid molecule described herein. BLAST protein searches can be performed with the XBLAST program parameters set, e.g., to score 50, wordlength=3 to obtain amino acid sequences homologous to a protein molecule described herein. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul SF et al., (1997) Nuc Acids Res 25: 3389-3402, which is herein incorporated by reference in its entirety. Alternatively, PSI BLAST can be used to perform an iterated search which detects distant relationships between molecules (Id.). When utilizing BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of the respective programs (e.g., of XBLAST and NBLAST) can be used (see, e.g., National Center for Biotechnology Information (NCBI) on the worldwide web, ncbi.nlm.nih.gov). Another specific, non-limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, 1988, CAB IOS 4: 11-17, which is herein incorporated by reference in its entirety. Such an algorithm is incorporated in the ALIGN program (version 2.0) which is part of the GCG sequence alignment software package. When utilizing the ALIGN program for comparing amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used. The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percent identity, typically only exact matches are counted.
As used herein, the terms “free targeted covalent inhibitor” or “free drug” refer to a targeted covalent inhibitor that is not covalently linked to a protein or peptide. Once the targeted covalent inhibitor is covalently linked to a protein or peptide, the targeted covalent inhibitor can be referred as a portion or fragment of the free target covalent inhibitor or drug. For example, the protein or fragment thereof can be the chemical fragment that is bonded to the cysteine residue of the peptide upon covalent reaction of the free drug with the cysteine residue of the peptide. As used herein the term “KRASG12C” refers to the KRAS protein (UniProt Accession No. P01116) with a G12C mutation, i.e., a cysteine at amino acid position 12. As used herein the term “KRASG12D” refers to the KRAS protein (UniProt Accession No. P01116) with a G12D mutation, i.e., an aspartic acid at amino acid position 12. As used herein the term “KRASG12R” refers to the KRAS protein (UniProt Accession No. P01116) with a G12R mutation, i.e., an arginine at amino acid position 12. As used herein the term “KRASG12S” refers to the KRAS protein (UniProt Accession No. P01116) with a G12S mutation, i.e., a serine at amino acid position 12.
In embodiments, the binding partners preferentially bind to (e.g., specifically bind to) the protein or peptide or a complex comprising the protein or peptide when covalently bound to the peptide conjugate, relative to the same protein or peptide that is not bound to the drug. Accordingly, binding partners described herein either do not detectably bind, or bind with a lower affinity, to the same protein or fragment thereof in the absence of the covalently attached molecule. In embodiments, the binding partners bind to the protein or peptide comprising the covalently attached drug with an affinity that is 10 - 10,000 fold, including all numbers and ranges of numbers from 10-10,000, greater than the affinity for the protein or peptide that does not comprise the covalently bound molecule. In this regard, and without intending to be bound by any particular theory, it is considered that the presence of the covalently bound molecule contributes to the epitope to which the binding partners bind with specificity. The term “specifically binds” refers to a molecule (e.g., an antibody or an antigen-binding portion thereof) that binds to an epitope or target or peptide-MHC complex with greater affinity, greater avidity, and/or greater duration to that epitope or target or peptide-MHC complex in a sample than it binds to another epitope or non-target compound or non-target or peptide-MHC complex (e.g., a structurally different antigen, a peptide-MHC complex with a different MHC and different peptide, a peptide-MHC complex with a different MHC and the same peptide or a peptide-MHC complex with the same MHC and different peptide). For example, a molecule (e.g., an antibody or an antigen-binding portion thereof) that specifically binds to an epitope or target or peptide-MHC complex can be an molecule (e.g., an antibody or an antigen-binding portion thereof) that binds this epitope or target or peptide-MHC complex with greater affinity, avidity, more readily, and/or with greater duration than it binds to other epitopes or targets or peptide-MHC complexes. In some embodiments, a molecule (e.g., an antibody or an antigen-binding portion thereof) that specifically binds to an epitope or target or peptide-MHC complex is a molecule (e.g., an antibody or an antigen-binding portion thereof) that binds to the epitope or target or peptide- MHC complex with at least 5-fold greater affinity than other epitopes or non-target compounds or non-target peptide-MHC complex, e.g., at least 5-fold, 10-fold, 100-fold, 1,000-fold, 10,000-fold, or greater affinity. A molecule (e.g., an antibody or an antigenbinding portion thereof) that specifically binds to a particular epitope or target or peptide- MHC complex can be exhibited, for example, by a molecule having an equilibrium dissociation constant KD for the epitope or target or peptide-MHC complex to which it binds of, e.g., 10-4 M or smaller, or 10 12 M, such as determined by, e.g., immunoassays, surface plasma resonance (e.g., Biacore™ assay), biolayer interferometry, or other assays known in the art. It will be recognized by one of skill in the art that an antibody that specifically binds to a target from one species may also specifically bind to orthologs of that target. In some embodiments, the extent of binding of a molecule (e.g., an antibody or an antigen-binding portion thereof) to an unrelated epitope or unrelated target or unrelated peptide-MHC complex is less than about 0.0001%, 0.001%, 0.01%, 0.1%, 1%, 10% or 20% of the binding of the antibody to the epitope or target or peptide-MHC complex as measured, e.g., by an immunoassay, surface plasma resonance (e.g., Biacore™ assay), biolayer interferometry, or other assay known in the art. Likewise, binding partners of this disclosure preferentially bind to the peptide conjugate relative to binding to the free drug. In embodiments, the binding partners bind to the peptide comprising the covalently attached drug (e.g., the peptide-conjugate/MHC complex) with an affinity that is 10-10,000 fold, including all numbers and ranges of numbers from 10-10,000, greater than the affinity for the free drug. In some embodiments, the interaction between the binding partner and the peptide-conjugate/MHC complex is not inhibited by the free drug. For example, the interaction between the binding partner and the peptide-conjugate/MHC complex is not inhibited by a lOOx, l,000x, 10,000x, 100,000x or more excess of the free drug.
In some embodiments, the polypeptide comprising the antigen-binding domain may bind to (i) a first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 100 nM, (ii) a second peptide conjugate/MHC complex with a KD of at most about 100 nM, and (iii) a third peptide conjugate/MHC complex with a KD of at most about 100 nM. The first peptide conjugate/MHC complex can comprise: a first peptide conjugate, wherein the first peptide conjugate of the first peptide conjugate/MHC complex is a first peptide covalently linked to a first targeted covalent inhibitor or fragment thereof; and a first MHC. The second peptide conjugate/MHC complex can comprise: a second peptide conjugate, wherein the second peptide conjugate of the second peptide conjugate/MHC complex is a second peptide covalently linked to a second targeted covalent inhibitor or fragment thereof; and a second MHC. The third peptide conjugate/MHC complex can comprise: a third peptide conjugate, wherein the third peptide conjugate of the third peptide conjugate/MHC complex is a third peptide covalently linked to a third targeted covalent inhibitor or fragment thereof; and a third MHC. In some embodiments, the first targeted covalent inhibitor, the second targeted covalent inhibitor, and the third targeted covalent inhibitor can be different. The first targeted covalent inhibitor can be adagrasib, and the second and third covalent inhibitors can be selected from the group consisting of sotorasib and divarasib. In some embodiments, the antigen-binding domain can bind to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 50 nM, (ii) the second peptide conjugate/MHC complex with a KD of at most about 50 nM, and (iii) the third peptide conjugate/MHC complex with a KD of at most about 50 nM. In some embodiments, the antigen-binding domain can bind to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 10 nM, (ii) the second peptide conjugate/MHC complex with a KD of at most about 10 nM, and (iii) the third peptide conjugate/MHC complex with a KD of at most about 10 nM.
In some embodiments, a polypeptide described herein may bind with greater affinity to a peptide conjugate/MHC complex comprising a peptide covalently linked to adagrasib compared to an affinity to a peptide conjugate/MHC complex comprising a peptide covalently linked to divarasib. In some embodiments, a polypeptide described herein may bind with greater affinity to a peptide conjugate/MHC complex comprising a peptide covalently linked to adagrasib compared to an affinity to a peptide conjugate/MHC complex comprising a peptide covalently linked to sotorasib.
In some embodiments, a binding partner described herein may bind to a first peptide conjugate/MHC complex (e.g., peptide conjugate with adagrasib covalently linked to the peptide) with a dissociation constant (KD) that can be at least about 2-fold higher, at least about 5-fold higher, at least about 10-fold higher, at least about 15-fold higher, at least about 20-fold higher, at least about 50-fold higher, at least about 100-fold higher, at least about 150-fold higher, at least about 250-fold higher, at least about 500-fold higher, at least about 1,000-fold higher, at least about 2,500-fold higher, at least about 5,000-fold higher, at least about 10,000-fold higher, at least about 50,000-fold higher, at least about 100,000-fold higher, or greater than about 100,000-fold higher than a dissociation constant (KD) resulting from the binding partner binding to a second conjugate/MHC complex (e.g., peptide conjugate with sotorasib covalently linked to the peptide) or a third conjugate/MHC complex (e.g., peptide conjugate with divarasib covalently linked to the peptide). In some embodiments, a binding partner described herein may bind to a first peptide conjugate/MHC complex (e.g., peptide conjugate with adagrasib covalently linked to the peptide) with a dissociation constant (KD) that can be at most about 100,000-fold higher, at most about 50,000-fold higher, at most about 10,000-fold higher, at most about 5,000-fold higher, at most about 2,500-fold higher, at most about 1,000-fold higher, at most about 500-fold higher, at most about 250-fold higher, at most about 150-fold higher, at most about 100-fold higher, at most about 50-fold higher, at most about 10-fold higher, at most about 5-fold higher, at most about 2-fold higher, or less than about 2-fold higher than a dissociation constant (KD) resulting from the binding partner binding to a second conjugate/MHC complex (e.g., peptide conjugate with sotorasib covalently linked to the peptide) or a third conjugate/MHC complex (e.g., peptide conjugate with divarasib covalently linked to the peptide).
In embodiments, the molecule that is covalently bound to form the peptide conjugate is a drug and may be any targeted covalent inhibitor (TCI), but the covalent drug need not necessarily inhibit the target peptide. In embodiments, the molecule reacts with a specific residue within the target protein. In embodiments, the molecule reacts at least in part with a segment of the protein or peptide that comprises a nucleophilic, or an electrophilic, residue. In embodiments, the segment of the protein or peptide to which the molecule reacts comprises any of Cys, Lys, Tyr, His, Ser, Thr, or Arg, the latter being described in Ziyang Zhang, Johannes Morstein, Andrew K. Ecker, Keelan Z. Guiley, and Kevan M. Shokat Journal of the American Chemical Society Article ASAP, DOI: 10.1021/jacs.2c05377, from which the disclosure is incorporated herein by reference. In embodiments, the protein or peptide comprises a selenocysteine. In embodiments, the targeted covalent inhibitor reacts with selenocysteine. In embodiments, the molecule reacts at least in part with a segment of the protein or peptide that comprises a wild type Cys, or a mutation of a residue to a Cys, and thus may be covalently attached by a so-called sulfur tether. In embodiments, the drug is any drug described in Ghosh AK, Samanta I, Mondal A, Liu WR. Covalent Inhibition in Drug Discovery. ChemMedChem. 2019;14(9):889-906. doi: 10.1002/cmdc.201900107, or in De Cesco, et al., European Journal of Medicinal Chemistry 138 (2017) 96el 14, or in Bauer, RA, Drug Discovery Today, Volume 20, Number 9, September 2015, from which the disclosures of compounds that covalently modify protein targets is incorporated herein by reference.
In non-limiting embodiments, any of said Asp, Cys, and Arg amino acids are present in the protein or peptide to which the molecule binds because the gene encoding the wild type protein has been mutated to encode a protein that includes one or a combination of the described residues. In non-limiting embodiments, the molecule binds to a protein or peptide that is correlated with a disease or condition, such as a cancer, an autoimmune disease, or other disease or disorder that is treated with a targeted covalent inhibitor. In embodiments, the target (e.g., the protein or peptide to which the molecule covalently binds) is a receptor, including but not necessarily limited to any receptor having a catalytically active segment. In embodiments, the drug binds to an enzyme that is not necessarily a receptor, including but not limited to any kinase. In embodiments, a protein target comprises a receptor with one or more activating mutations, which promote ligand-independent enzyme activity.
In embodiments, the molecule targets and thus covalently binds to an amino acid sequence present within any of the following proteins and/or variants thereof, which may or may not comprise a mutation, such as a mutation that is related to a particular condition, including but not limited to any type of cancer. In embodiments, the protein is any protein described in Visscher M, et al., Covalent targeting of acquired cysteines in cancer. Curr Opin Chem Biol. 2016;30:61-67. doi: 10.1016/j.cbpa.2015.11.004, from which the description is incorporated herein by reference. Visscher et al. also teaches methods for identifying disease- associated mutated genes that introduces a Cys residue suitable for covalent modification. In embodiments, the protein is KRAS, Bruton's tyrosine kinase (BTK), any member of the epidermal growth factor receptor (EGFR) family, also referred to as the ERBB family, including but not limited to EGFR (ERBB1), HER2/NEU (ERBB2), HER3 (ERBB3), and HER4 (ERBB4); a fibroblast growth factor receptor (FGFR); the receptor kinase known in the art as MET, BRAF, a cyclin-dependent kinase (CDK); Acetyl Choline Esterase (ACHE); TP53, IDH1, GNAS, FBXW7, CTNNB1, DNMT3A, any cathepsin, including cathepsin B, C, F, H, K, L, O, S, V, W and X; any caspase; any protein involved in obesity, such as Pancreatic lipase and METAP2, or any Cancer Testis Antigen. In embodiments, the drug targets and therefore covalently binds to any viral protein, including but not limited to a polymerase, including any viral DNA polymerase, RNA polymerase, reverse transcriptase, or RNA-dependent RNA polymerase, or a viral protein that is required, for example, viral cell entry, or a protein encoded by any a transposable element. In embodiments, the drug targets EGFR and may be selected from PD168393, PF00299804 (dacomitinib), EKB569 (pelitinib), afatinib, WZ4002, osimertinib (formerly known as AZD9291), PF-06459988, nazartinib, naquotinib, olmutinib, avitinib, and rociletinib, neratinib, pyrotinib, poziotinib, and derivatives thereof. In embodiments, the drug targets Bruton’s tyrosine kinase (BTK), and may be selected from ibrutinib, acalabrutinib, zanubrutinib, CHMFL-BTK-11, ONO/GS-405, PRN1008, and CC-292. In embodiments, the drug targets any p90 ribosomal S6 kinase (RSK), and may be selected from fluoromethylketone (FMK) and dimethyl fumarate. In embodiments, the drug targets any FGFR, and may be selected from FIIN-1, FIIN-2, FIIN-3, BGJ398, AZD4547, PRN1371, FGF401. In an embodiment, the targeted covalent inhibitor targets an E3 ligase, such as RNF4, HOIP, RSP5, SMURF1, E6AP, HUWE1, and NEDD4-1. In an embodiment, the targeted covalent inhibitor targets a DDB1- and CUL4- associated factor (DCAF), such as DCAF1 or DCAF15. In an embodiment, the targeted covalent inhibitor targets any cancer testis antigen, any endogenous retroviral protein, a long interspersed element-1 (LINE-1), or a short interspersed element (SINE). In an embodiment, the targeted covalent inhibitor targets a short interspersed element that is optionally Alu. In an embodiment, the targeted covalent inhibitor is iniparib, abiraterone, carfilzomib, afatinib, or neratinib. In some embodiments, the targeted covalent inhibitor can be a targeted covalent agent. The targeted covalent agent may not inhibit the cancer testis antigen, endogenous retroviral protein, long interspersed element- 1 (LINE-1), or short interspersed element (SINE).
In embodiments, the molecule that becomes covalently bound to form the peptide conjugate targets any RAS oncogene protein product, including but not necessarily limited to HRAS, NRAS, KRAS4A, and KRAS4B. The amino acid sequences of RAS proteins are known in the art, and residue numbering is identical for the relevant part of all RAS isotypes that are discussed in this disclosure for which the amino acid sequence is available from, for example, UniProt P01116, from which the amino acid sequence is incorporated herein as of the effective filing date of this application or patent. The G12 position is numbered according to the known amino acid sequence, regardless of whether or not the G12 is the twelfth amino acid in an express RAS peptide sequence of this disclosure.
In one embodiment, the molecule covalently binds to a KRAS protein or peptide that comprises a mutation. In embodiments, the mutation is at least one of KRAS residues 12, 13, or 61. Reference to any drug herein includes its name in capitalized and un-capitalized form.
In some embodiments, the drug targets a KRAS protein comprising a KRAS G12C mutation. In some embodiments, the drug targets a KRAS protein comprising a KRAS G12D mutation. In some embodiments, the drug targets a KRAS protein comprising a KRAS G12R mutation. In some embodiments, the drug targets a KRAS protein comprising a KRAS G12S mutation. In non-limiting embodiments, the drug that targets a KRAS protein is selected from 2E07, 6H05, SML-8-73-1, MRTX849 (e.g., adagrasib), JNJ74699157, LY3499446, ARS- 853, ARS-1620, ARS-3284, GDC-6036 (e.g., divarasib), D-1553, JDQ443, RMC-6291, RMC-9805, BI 1823911, MRTX1257, AMG-510 (e.g., sotorasib), or derivatives thereof. Examples of additional compounds that can covalently target a KRAS peptide containing a G12C mutation can be found in Internal Application No. PCT/IB2019/050993, Internal Application No. PCT/EP2018/083853, and U.S. Application No. US16/917,128, each of which is incorporated herein by reference in its entirety. In an embodiment the drug comprises a proteolysis targeting chimera (PROTAC) derivative of a covalent drug, a non- limiting description of which is available in doi: 10.1021/acscentsci.0c00411, from which the description of PROTACs is incorporated herein by reference. In embodiments, the PROTAC is LC-1 or LC-2. In embodiments, the disclosure relates to an autophagy-mediated degrader, referred to as an AUTAC, as described in doi.org/10.1080/15548627.2020.1718362, from which the description of AUTACs is incorporated herein by reference.
The peptide conjugate described herein can be formed by the covalent reaction of a free targeted covalent inhibitor with a KRAS peptide. The peptide conjugate can be formed by the covalent reaction of a free targeted covalent inhibitor with a KRASG12C peptide, a KRASG12D peptide, a KRASG12R peptide, or a KRASG12S peptide. The free targeted covalent inhibitor can be any free targeted covalent inhibitor described herein. For example, the free targeted covalent inhibitor can be AMG-510 (e.g., sotorasib), MRTX849 (e.g., adagrasib), or GDC6036 (e.g., divarasib). In some cases, the peptide conjugate can be formed by the covalent reaction of AMG-510 with a KRASG12C peptide. The peptide can comprise or consist of the amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV. In some cases, the targeted covalent inhibitor may be beta-lactones G12Si- 1, G12Si-2, G12Si-3, G12Si-4 or G12Si-5.
The antigen-binding domain of the polypeptide described herein or the binding partner described herein can bind to a peptide conjugate/MHC complex presented by different HLAs. For example, in certain embodiments, the antigen-binding domain can have specificity to: (i) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01, (ii) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01, or (iii) both. In certain other embodiments, the antigen-binding domain can have specificity to: (i) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01, (ii) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01, or (iii) both. In other embodiments, the antigen-binding domain can have specificity to: (i) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01, (ii) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01, (iii) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01, (iv) a peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01, or (v) any combination of (i) - (iv) (e.g., (i) and (ii); or (i), (ii), and (iv)), or (vi) all of (i)-(iv). In certain specific embodiments, the polypeptide binds to: (i) a peptide conjugate/ HLA-A*03:01 MHC complex containing a peptide consisting of the amino acid sequence VVGACGVGK, (ii) a peptide conjugate/ HLA-A*03:01 MHC complex containing a peptide consisting of the amino acid sequence VVVGACGVGK, (iii) a peptide conjugate/ HLA-A* 11:01 MHC complex containing a peptide consisting of the amino acid sequence VVVGACGVGK, and/or (iv) a peptide conjugate/ HLA-A*02:01 MHC complex containing a peptide consisting of the amino acid sequence KLVVVGACGV. In still other embodiments, the polypeptide binds to: (i) a peptide conjugate/ HLA-A*03:01 MHC complex containing a peptide consisting of the amino acid sequence VVGACGVGK and a peptide conjugate/ HLA- A*03:01 MHC complex containing a peptide consisting of the amino acid sequence VVVGACGVGK; (ii) a peptide conjugate/ HLA-A*03:01 MHC complex containing a peptide consisting of the amino acid sequence VVVGACGVGK and a peptide conjugate/ HLA-A* 11:01 MHC complex containing a peptide consisting of the amino acid sequence VVVGACGVGK; (iii) a peptide conjugate/ HLA-A* 02:01 MHC complex containing a peptide consisting of the amino acid sequence KLVVVGACGV; or any combination of (i)- (iii) (e.g., (i) and (ii); or (i) and (iii)); or all of (i) - (iii). The covalent inhibitor that targets the peptide conjugate/MHC complex can have a chemical structure comprising C-Rl, where C is a chemical fragment linked to R1 of any compound illustrated below.
R1 of the C-Rl chemical structure of the covalent inhibitor can be any of the structures illustrated below. Compound (1) designates AMG-510 (sotorasib), Compound (2) designates MRTX849 (adagrasib), and Compound (3) designates GDC6036 (divarasib).
The targeted covalent inhibitor with the structure comprising C-Rl can form a covalent bond to several different amino acid residues on the peptide. For example, the targeted covalent inhibitor can form a covalent bond to the cysteine residue in a peptide comprising the amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV. In other embodiments, the targeted covalent inhibitor can form a covalent bond to the aspartic acid residue, the serine residue, or the arginine residue in a peptide comprising the amino acid sequence of VVVGADGVGK, VVGADGVGK, or KLVVVGADGV. In some embodiments, the antigen-binding domain of the polypeptide recognizes the C portion of the targeted covalent inhibitor with the structure comprising C-Rl of the peptide conjugate/MHC complex. In certain other embodiments, the antigen-binding domain of the polypeptide recognizes C portion but not R1 portion of the targeted covalent inhibitor of the peptide conjugate/MHC complex.
Small molecules (e.g., targeted covalent inhibitors) having an electrophilic warhead group can undergo covalent reaction with a cysteine residue of a peptide to form a peptide- small molecule conjugate. This type of reaction is illustrated in the following scheme for AMG-510 (sotorasib).
The table below lists targeted covalent inhibitors, along with the chemical structure of the fragment (“chemical fragment”) that is bonded to the cysteine residue of the peptide upon covalent reaction of the drug with the cysteine. In an embodiment, the binding partners disclosed herein bind to a peptide conjugate/MHC complex comprising a peptide conjugated to a chemical fragment in Table A below and an MHC. Table A. Structures of targeted covalent inhibitors and the chemical fragments after covalent reaction with a cysteine residue.
In a non-limiting embodiment, the binding partner binds with specificity to a site comprising a neoantigen that includes a covalently linked small molecule drug or other covalently linked molecule as a component of an antigen in a specific MHC context. In an aspect, provided herein is a binding partner that specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of a targeted covalent inhibitor with a peptide. In an embodiment, the binding partner binds to the peptide conjugate/MHC complex with a greater affinity than to the peptide or free targeted covalent inhibitor.
In an aspect, the present disclosure provides a binding partner that specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate/MHC complex comprises: (a) a peptide conjugate formed by the covalent reaction of a targeted covalent inhibitor or fragment thereof with a peptide; and(b) an MHC.
In some embodiments, the binding partner binds to the peptide conjugate/MHC complex with a greater affinity than to the peptide or free targeted covalent inhibitor. In some embodiments, the affinity of the binding partner for the peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the binding partner for the peptide or free targeted covalent inhibitor.
In some embodiments, the MHC is a human leukocyte antigen (HLA), optionally wherein the HLA is an HLA-A, HLA-B, or HLA-C. In some embodiments, the HLA is HLA- A* 02 : 01 , HL A- A* 03 : 01 , or HLA-A* 11 :01.
In some embodiments, the peptide comprises a nucleophilic or an electrophilic residue, said residue optionally being one of cysteine, aspartic acid, or arginine.
In some embodiments, the peptide comprises a cysteine residue.
In some embodiments, the peptide conjugate is formed by a covalent reaction between the targeted covalent inhibitor and a cysteine residue in the peptide. In some embodiments, the peptide is a segment of a protein that is associated with a cancer, optionally wherein the protein is encoded by a gene that is mutated in a cancer.
In some embodiments, the peptide is a segment of an enzyme, and wherein the targeted covalent inhibitor is an inhibitor of the enzyme. In some embodiments, the enzyme is a kinase or a GTPase.
In some embodiments, the peptide is or is derived from KRAS.
In some embodiments, the peptide comprises a segment of KRASG12C, KRASG12D, KRASG12S, or KRASG12R.
In some embodiments, the peptide comprises the amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV.
In some embodiments, the targeted covalent inhibitor is (i) a tri-complex KRASG12C inhibitor or a KRASG12C degrader, (ii) a tri-complex KRASG12D inhibitor or a KRASG12D degrader, (iii) a tri-complex KRASG12R inhibitor or a KRASG12R degrader, or (iv) a tri- complex KRASG12S inhibitor or a KRASG12S degrader.
In some embodiments, the peptide comprises the KRASG12C mutation, and the targeted covalent inhibitor is a KRASG12C inhibitor. In some embodiments, the peptide comprises the KRASG12D mutation, and the targeted covalent inhibitor is a KRASG12D inhibitor. In some embodiments, the peptide comprises the KRASG12R mutation, and the targeted covalent inhibitor is a KRASG12R inhibitor. In some embodiments, the peptide comprises the KRASG12S mutation, and the targeted covalent inhibitor is a KRASG12S inhibitor.
In some embodiments, the targeted covalent inhibitor is sotorasib (e.g., AMG-510), adagrasib (e.g., MRTX849), divarasib (e.g., GDC6036), or any combination thereof.
In some cases, the targeted covalent inhibitor is 2E07, 6H05, SML-8-73-1, MRTX849, JNJ74699157, LY3499446, ARS-853, ARS-1620, ARS-3284, GDC-6036, D- 1553, JDQ443, BI 1823911, or derivatives thereof.
In some embodiments, the peptide is a full length protein that is processed in the cell after the covalent reaction with the targeted covalent inhibitor, such that smaller peptide fragments are produced.
In some embodiments, the peptide conjugate comprises a compound selected from the group consisting of compounds (1) and (2):
covalently bonded to the cysteine residue in a peptide comprising the amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV.
In some embodiments, the MHC is HLA-A*02:01, HLA-A*03:01, and/or HLA- A*l l:01.
In some embodiments, the peptide comprises the amino acid sequence of VVVGACGVGK or VVGACGVGK and the MHC is HLA-A*03 :01 or HLA-A* 11 :01. In some embodiments, the peptide comprises the amino acid sequence of KLVVVGACGV and the MHC is HLA-A*02:0L
In some embodiments, the MHC is HLA-A*01:01.
In some embodiments, the binding partner specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of AMG-510 or MRTX849 with a KRASG12C peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL). In some embodiments, the VH comprises: (i) a HC CDR1 comprising the amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising the amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising the amino acid sequence of LWASGLDY. In some embodiments, the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V. In some embodiments, the binding partner specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of AMG-510 or MRTX849 with a KRASG12C peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL). In some embodiments, the VH comprises: (i) a HC CDR1 comprising the amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising the amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising the amino acid sequence of SYYGFWQALWALDY. In some embodiments, the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In some embodiments, the binding partner specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of AMG-510 or MRTX849 with a KRASG12C peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL). In some embodiments, the VH comprises: (i) a HC CDR1 comprising the amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising the amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising the amino acid sequence of GYYYPYYAMDY. In some embodiments, the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In some embodiments, the binding partner specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of AMG-510 or MRTX849 with a KRASG12C peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL). In some embodiments, the VH comprises: (i) a HC CDR1 comprising the amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising the amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising the amino acid sequence of GYYYPYYAMDY. In some embodiments, the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In some embodiments, the binding partner specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of AMG-510 or MRTX849 with a KRASG12C peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL). In some embodiments, the VH comprises: (i) a HC CDR1 comprising the amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising the amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising the amino acid sequence of X1X2X3X4X5MDY, wherein Xi is F or G, X2 is Q or Y, X3 is W or G, X4 is Y or W, and X5 is A or G. In some embodiments, the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In some embodiments, the binding partner specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of AMG-510 or MRTX849 with a KRASG12C peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL). In some embodiments, the VH comprises: (i) a HC CDR1 comprising the amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising the amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising the amino acid sequence of SGYYSSHWYLQSWYQAMDY. In some embodiments, the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In some embodiments, the binding partner specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of AMG-510 or MRTX849 with a KRASG12C peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL). In some embodiments, the VH comprises: (i) a HC CDR1 comprising the amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising the amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising the amino acid sequence of SGYYSSHWYLQSWYQAMDY. In some embodiments, the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
In some embodiments, the binding partner specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of AMG-510 or MRTX849 with a KRASG12C peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL). In some embodiments, the VH comprises: (i) a HC CDR1 comprising the amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising the amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising the amino acid sequence of HYSEKWWGWYTMYIDAMDY. In some embodiments, the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In some embodiments, the binding partner specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of AMG-510 or MRTX849 with a KRASG12C peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL). In some embodiments, the VH comprises: (i) a HC CDR1 comprising the amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising the amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising the amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L. In some embodiments, the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In some embodiments, the binding partner specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of AMG-510 or MRTX849 with a KRASG12C peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL). In some embodiments, the VH comprises: (i) a HC CDR1 comprising the amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising the amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising the amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L. In some embodiments, the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQXiX2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
In some embodiments, the binding partner specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of AMG-510 or MRTX849 with a KRASG12C peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL). In some embodiments, the VH comprises: (i) a HC CDR1 comprising the amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising the amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising the amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L. In some embodiments, the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In some embodiments, the binding partner specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of AMG-510 or MRTX849 with a KRASG12C peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL). In some embodiments, the VH comprises: (i) a HC CDR1 comprising the amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising the amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or (iii) a HC CDR3 comprising the amino acid sequence of SSRQYYHSQVEPPMAMDY. In some embodiments, the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In some embodiments, the binding partner specifically binds to a peptide conjugate/MHC complex, wherein the peptide conjugate is formed by the covalent reaction of AMG-510 or MRTX849 with a KRASG12C peptide, wherein the binding partner comprises the HC CDR1, HC CDR2, and HC CDR3 amino acid sequences of a VH amino acid sequence and/or the LC CDR1, LC CDR2, and LC CDR3 amino acid sequences of a VL amino acid sequence of a binding partner selected from the group consisting of RM 001, RM_002, RM_003, RM_004, RM_005, RM_006, RM_007, RM_008, RM_009, RM_010, RM_011, RM_012, RM_013, RM_014, RM_015, RM_016, RM_017, RM_018, RM_019, RM_020, RM_021, RM_022, , RM_023, RM_024, RM_025, RM_026, RM_027, RM_028 , RM-029, RM-030, RM_031, and RM_032; or a variant thereof comprising 1-5 amino acid changes in one or more of the CDR amino acid sequences.
In some embodiments, the binding partner comprises the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and/or LC CDR3 amino acid sequences of a binding partner selected from the group consisting of RM_001, RM_002, RM_003, RM_004, RM_005, RM_006, RM_007, RM_008, RM_009, RM_010, RM_011, RM_012, RM_013, RM_014, RM_015, RM_016, RM_017, RM_018, RM_019, RM_020, RM_021, RM_022, , RM_023, RM_024, RM_025, RM_026, RM_027, RM_028 , RM_029, RM_030, RM_031, and RM-032 (Tables A, B, and C), or a variant thereof comprising 1-5 amino acid changes in one or more of the CDR amino acid sequences.
In some embodiments, the binding partner comprises a VH and/or a VL amino acid sequence that is 90%, 95%, or 100% identical to the following VH and VL sequences:
RM-001
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSG VPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTISSSSIHWVRQAPGKGLEWVASISSYYG STSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWGQGTL VTVSS
RM_002
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSG VP
SRFSGSRSGTDFTL TISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSYYG STSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWGQGTL VTVSS
RM_003
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSG VPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSSSGS TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALWALDY WGQGTL VTVSS
RM_004
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSG VPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISSSSG
STSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALWALDY WGQGTLVTVSS
RM_005
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSG
VPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSYY
GSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS
RM_006
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSG
VPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISPYY
GSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS
RM_007
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTL TISSLQPEDFATYYCQQTSWYHSLITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSYYGS
TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWGQG TLVTVSS
RM_008
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVASISSYYG
STYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWGQG TLVTVSS
RM_009
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSYYGS
TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWGQG TLVTVSS
RM_010
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSWLYWLVTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSSSGST
SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFQWYAMDYWGQGTLVTV SS
RM_011
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTISSYSIHWVRQAPGKGLEWVAYISSYSGY
TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYGWGMDYWGQGTLV TVSS
RM_012
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTISYSSIHWVRQAPGKGLEWVAYISSSSGY
TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQSWYQA MDYWGQGTLVTVSS
RM_013
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTL TISSLQPEDFATYYCQQASYGPITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSYYGS
TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQSWYQA MDYWGQGTLVTVSS
RM_014
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTL TISSLQPEDFATYYCQQWWSSSQLITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVASISSYYGS
TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARHYSEKWWGWYTMYID AMDYWGQGTLVTVSS
RM_015
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG
YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGT LVTVSS
RM_016
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY
TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGTL VTVSS
RM_017
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG
YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGT L VTVSS
RM_018
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPYSG
YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGT
L VTVSS
RM_019
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISPYSSY
TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGTL VTVSS
RM_020
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG
YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGT L VTVSS
RM_021
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY
TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTL VTVSS
RM_022
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTL VTVSS
RM_023
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSYYEELITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY
TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTL VTVSS
RM_024
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQAYSDPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY
TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTL VTVSS
RM 025
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY
TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTL VTVSS
RM_026
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSGSYLLITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY
TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTL VTVSS
RM_027
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQADYEFGLITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY
TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTL VTVSS
RM_028
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMAM DYWGQGTLVTVSS
RM_029
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVAYISSSYG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS
RM_030
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMAM DYWGQGTLVTVSS
RM_031
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVAYISSSYG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS
RM_032
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPYSG YTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYLGMDYWGQGT LVTVSS
In some embodiments, the binding partner has a higher affinity for the peptide conjugate/MHC complex comprising a first HLA than for the peptide conjugate/MHC complex comprising a second HLA.
In some embodiments, the first and second HLAs are each selected from the group consisting of HL A-A* 02:01, HLA-A*03:01, and/or HLA-A* 11 :01.
In some embodiments, the binding partner is an intact antibody, a bispecific antibody, a multispecific antibody, an antigen-binding (Fab) fragment, an Fab’ fragment, an (Fab’)2 fragment, an Fd, an Fv, a dAb, a single domain fragment or single monomeric variable antibody domain, a single-chain Diabody (scDb), a diabody (Db), a dual-affinity retargeting (DART) molecule, a single-chain variable fragment (scFv), a camelid antibody, a bi-specific T-cell engager (BiTE), bispecific killer cell engager (BiKE), CrossMab, a tri-specific binding partner, a chimeric antigen receptor (CAR), a monobody (e.g., adnectin), a DARPin, an anticalin, an affibody, or an affimer. In some embodiments, the binding partner is bispecific. In some embodiments, the binding partner specifically binds to the peptide conjugate/MHC complex and a T cell antigen. In some embodiments, the binding partner specifically binds to the peptide conjugate/MHC complex and human CD3.
In some embodiments, the binding partner comprises the VH and/or VL amino acid sequences of
UCHT1:
VH:EVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWMGLINPYKGVS TYNQKFKDKATLTVDKSSSTAYMELLSLTSEDSAVYYCARSGYYGDSDWYFDVWGQGTTL TVSS VL:DIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHSGVPSK FSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIK.
In some embodiments, the binding partner is a single-chain Diabody (scDb) and comprises a sequence that is at least 90% identical to any of the following sequences:
RM 001 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTVGGGGSEVQL VESGGGLVQPGGSLRLSCAASGFTISSSSIHWVRQAPGKGLEWVASISSYYGSTSYADSVKGR
FTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWGQGTLVTVSSGGGGSGGGGS
GGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHSG VPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVESG
GGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTIS
ADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 002 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTVGGGGSEVQL VESGGGLVQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSYYGSTSYADSVKG
RFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWGQGTLVTVSSGGGGSGGGG SGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHSG
VPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVESG GGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTIS
ADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 003 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRTVGGGGSEVQL VESGGGLVQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSSSGSTSYADSVKGR
FTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALWALDYWGQGTLVTVSSGGGG SGGGGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYT
SRLHSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEV QLVESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVK
GRFTISADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 004 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRTVGGGGSEVQL
VESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISSSSGSTSYADSVKGR
FTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALWALDYWGQGTLVTVSSGGGG
SGGGGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYT
SRLHSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEV
QLVESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVK
GRFTISADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 005 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTVGGGGSEVQLVE
SGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSYYGSTYYADSVKGRF
TISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWGQGTLVTVSSGGGGSGG
GGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRL
HSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLV
ESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRF
TISADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 006 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTVGGGGSEVQLVE
SGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISPYYGSTYYADSVKGRF
TISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWGQGTLVTVSSGGGGSGG
GGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRL
HSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLV
ESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRF
TISADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 007 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQTSWYHSLITFGQGTKVEIKRTVGGGGSEVQLV
ESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSYYGSTYYADSVKGR
FTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWGQGTLVTVSSGGGGSGG
GGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRL
HSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLV
ESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRF
TISADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 008 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTVGGGGSEVQLVE
SGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVASISSYYGSTYYADSVKGRF
TISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWGQGTLVTVSSGGGGSGG
GGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRL
HSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLV
ESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRF
TISADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS RM 009 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTVGGGGSEVQLV
ESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSYYGSTYYADSVKGR
FTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWGQGTLVTVSSGGGGSGG
GGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRL
HSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLV
ESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRF
TISADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 010 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWLVTFGQGTKVEIKRTVGGGGSEVQL
VESGGGLVQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSSSGSTSYADSVKGR
FTISADTSKNTAYLQMNSLRAEDTAVYYCARFQWYAMDYWGQGTLVTVSSGGGGSGGGGS
GGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHSG
VPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVESG
GGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTIS
ADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM Oil UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTVGGGGSEVQL
VESGGGLVQPGGSLRLSCAASGFTISSYSIHWVRQAPGKGLEWVAYISSYSGYTSYADSVKG
RFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYGWGMDYWGQGTLVTVSSGGGGSGGG
GSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHS
GVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVES
GGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTI
SADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 012 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTVGGGGSEVQLV
ESGGGLVQPGGSLRLSCAASGFTISYSSIHWVRQAPGKGLEWVAYISSSSGYTSYADSVKGRF
TISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQSWYQAMDYWGQGTLVTVS
SGGGGSGGGGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKL
LIYYTSRLHSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGG
GSEVQLVESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYA
DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 013 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYGPITFGQGTKVEIKRTVGGGGSEVQLVES
GGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSYYGSTYYADSVKGRFT
ISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQSWYQAMDYWGQGTLVTVSS
GGGGSGGGGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLL
IYYTSRLHSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGG
SEVQLVESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYAD
SVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 014 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWSSSQLITFGQGTKVEIKRTVGGGGSEVQL
VESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVASISSYYGSTYYADSVKG RFTISADTSKNTAYLQMNSLRAEDTAVYYCARHYSEKWWGWYTMYIDAMDYWGQGTLVT VSSGGGGSGGGGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTV KLLIYYTSRLHSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKG GGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTS YADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 015 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTVGGGGSEVQLVE SGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSGYTSYADSVKGRF TISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGTLVTVSSGGGGSGGGG SGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHSG VPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVESG GGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTIS ADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 016 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTVGGGGSEVQLVE SGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSYTSYADSVKGRF TISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGTLVTVSSGGGGSGGGG SGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHSG VPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVESG GGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTIS ADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 017 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTVGGGGSEVQLV ESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSGYTSYADSVKGR FTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGTLVTVSSGGGGSGGG GSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHS GVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVES GGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTI SADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 018 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTVGGGGSEVQLV ESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPYSGYTSYADSVKGR FTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGTLVTVSSGGGGSGGG GSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHS GVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVES GGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTI SADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 019 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTVGGGGSEVQLV ESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISPYSSYTSYADSVKGRF TISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGTLVTVSSGGGGSGGGG SGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHSG VPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVESG GGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTIS
ADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 020 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTVGGGGSEVQLV
ESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSGYTSYADSVKGR
FTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGTLVTVSSGGGGSGGG
GSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHS
GVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVES
GGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTI
SADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 021 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTVGGGGSEVQLV
ESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSYTSYADSVKGR
FTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTLVTVSSGGGGSGGG
GSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHS
GVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVES
GGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTI
SADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 022 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTVGGGGSEVQLV
ESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSYTSYADSVKGR
FTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTLVTVSSGGGGSGGG
GSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHS
GVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVES
GGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTI
SADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 023 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSYYEELITFGQGTKVEIKRTVGGGGSEVQLV
ESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSYTSYADSVKGR
FTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTLVTVSSGGGGSGGG
GSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHS
GVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVES
GGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTI
SADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 024 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQAYSDPLTFGQGTKVEIKRTVGGGGSEVQLVES
GGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSYTSYADSVKGRFTI
SADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTLVTVSSGGGGSGGGGS
GGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHSG
VPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVESG
GGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTIS ADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 025 UCHT1 scDb DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTVGGGGSEVQLVES GGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSYTSYADSVKGRFTI SADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTLVTVSSGGGGSGGGGS GGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHSG
VPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVESG GGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTIS ADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 026 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSGSYLLITFGQGTKVEIKRTVGGGGSEVQLVE SGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSYTSYADSVKGRF TISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTLVTVSSGGGGSGGGG SGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHSG VPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVESG
GGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTIS ADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 027 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQADYEFGLITFGQGTKVEIKRTVGGGGSEVQLV ESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSYTSYADSVKGR FTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQGTLVTVSSGGGGSGGG GSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHS
GVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVES GGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTI SADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 028 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTVGGGGSEVQLV ESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVAYISSSYGYTSYADSVKGR FTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMAMDYWGQGTLVTVSS GGGGSGGGGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLL
IYYTSRLHSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGG SEVQLVESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYAD SVKGRFT1SADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 029 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTVGGGGSEVQLV ESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVAYISSSYGYTSYADSVKGR FTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMAMDYWGQGTLVTVSS GGGGSGGGGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLL
IYYTSRLHSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGG SEVQLVESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYAD SVKGRFT1SADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 030 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTVGGGGSEVQLV ESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYISSSYGYTSYADSVKGR FTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMAMDYWGQGTLVTVSS GGGGSGGGGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLL
IYYTSRLHSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGG SEVQLVESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYAD SVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 031 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTVGGGGSEVQLV
ESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVAYISSSYGYTSYADSVKGR
FTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMAMDYWGQGTLVTVSS
GGGGSGGGGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLL
IYYTSRLHSGVPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGG
SEVQLVESGGGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYAD
SVKGRFT1SADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
RM 032 UCHT1 scDb
DIVRSDIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV
PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTVGGGGSEVQLVES
GGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPYSGYTYYADSVKGRFT
ISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYLGMDYWGQGTLVTVSSGGGGSGGGGS
GGGGSDIQMTQTTSSLSASLGDRVTISCRASQDIRNYLNWYQQKPDGTVKLLIYYTSRLHSG
VPSKFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPWTFAGGTKLEIKGGGGSEVQLVESG
GGLVQPGGSLRLSCAASGFTISSSYIHWVRQAPGKGLEWVAYISPSYGSTSYADSVKGRFTIS ADTSKNTAYLQMNSLRAEDTAVYYCAREYVTMALDYWGQGTLVTVSS
In some embodiments, the binding partner comprises a heavy chain constant region selected from the group consisting of human IgM, IgGi, IgG2, IgGs, IgG4, IgAi, and IgA2.
In some embodiments, the heavy chain constant region comprises one or more amino acid substitutions, deletions, or additions in the Fc region.
In some embodiments, the binding partner comprises a human kappa light chain constant region or a human lambda light chain constant region.
In some embodiments, the binding partner is in a CrossMab format.
In some embodiments, the binding partner is conjugated to a detectable label, a chemotherapeutic agent, a radioisotope, or a toxin.
In some embodiments, the binding partner is comprised within a chimeric antigen receptor.
In some embodiments, the binding partner is expressed by a T cell, killer macrophage, neutrophil, or natural killer cell.
In some embodiments, binding of the binding partner to the peptide conjugate/MHC complex is not inhibited by free targeted covalent inhibitor.
In another aspect, the present disclosure provides a complex comprising a binding partner disclosed herein and the peptide conjugate/MHC complex. In another aspect, the present disclosure provides a polynucleotide encoding the binding partner of disclosed herein.
In another aspect, the present disclosure provides a polynucleotide encoding a heavy chain variable region and/or a light chain variable region of the binding partner disclosed herein.
In another aspect, the present disclosure provides a vector comprising the polynucleotide disclosed herein.
In some embodiments, the vector is a viral vector. In some embodiments, the viral vector is an adenoviral vector, lenti viral vector, retroviral vector, or adeno-associated viral vector.
In another aspect, the present disclosure provides a recombinant host cell comprising (a) the polynucleotide disclosed herein; (b) the vector disclosed herein; (c) a first polynucleotide encoding a VH or a heavy chain of the binding partner disclosed herein, and a second polynucleotide encoding a VL or a light chain of the binding partner disclosed herein; or (d) a first vector comprising a first polynucleotide encoding a VH or a heavy chain of the binding partner disclosed herein, and a second vector comprising a second polynucleotide encoding a VL or a light chain of the binding partner disclosed herein.
In another aspect, the present disclosure provides a pharmaceutical composition comprising the binding partner disclosed herein, the polynucleotide disclosed herein, the vector disclosed herein, or the host cell disclosed herein and a pharmaceutically acceptable carrier or excipient.
In another aspect, the present disclosure provides a method of producing a binding partner, the method comprising culturing the host cell disclosed herein under suitable conditions so that the polynucleotide is expressed and the binding partner is produced.
In another aspect, the present disclosure provides a eukaryotic cell comprising the polynucleotide disclosed herein or the vector disclosed herein. In some embodiments, the cell is optionally a totipotent, multipotent, or pluripotent stem cell. In some embodiments, the stem cell has an induced stem cell phenotype, or wherein the cell is optionally a leukocyte, optionally a CD4+ T cell, optionally a CD8+ T cell, optionally a y5 T cell, optionally a natural killer cell or a macrophage.
In another aspect, the present disclosure provides a method comprising administering to an individual in need thereof the binding partner disclosed herein, the polynucleotide disclosed herein, the vector disclosed herein, the pharmaceutical composition disclosed herein, or the cell disclosed herein.
In another aspect, the present disclosure provides a method for generating a peptide conjugate/MHC complex, the method comprising contacting a cell with a targeted covalent inhibitor, and isolating the peptide conjugate/MHC complex. In some embodiments, the method comprises identifying the peptide conjugate/MHC complex.
In another aspect, the present disclosure provides a cell free peptide conjugate/MHC complex comprising: (a) an isolated peptide conjugate formed by the covalent reaction of a targeted covalent inhibitor with a peptide; and (b) an MHC.
In some embodiments, the peptide comprises a segment of KRASG12C.
In some embodiments, the targeted covalent inhibitor is sotorasib (e.g., AMG-510), adagrasib (e.g., MRTX849), divarasib (e.g., GDC6036), or any combination thereof.
In another aspect, the present disclosure provides a cell free peptide conjugate/MHC complex comprising (a) a compound selected from the group consisting of compounds 1 and 2, covalently bonded to a cysteine residue in a peptide comprising the amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV; and (b) an MHC.
In some embodiments, the MHC is an HLA, optionally wherein the HLA is HLA- A* 02 : 01 , HL A- A* 03 : 01 , or HL A- A* 11 :01.
In another aspect, the present disclosure provides a recombinant cell or particle comprising on its outer surface the cell free peptide conjugate/MHC complex disclosed herein.
In another aspect, the present disclosure provides a method of identifying one or more binding partners that bind with specificity to a peptide conjugate presented by an HLA, the method comprising contacting the cell-free peptide conjugate/MHC complex disclosed herein with a plurality of binding partners, and selecting one or more binding partners that bind with specificity to the cell-free peptide conjugate/MHC complex. In some embodiments, the method further comprises determining the sequence of the one or more selected binding partners. In some embodiments, the method further comprises producing the one or more selected binding partners.
In another aspect, the present disclosure provides a method for identifying a binding partner that specifically binds to a peptide conjugate presented by two or more HLAs, the method comprising providing a sample of cells from a subject who has been treated with a targeted covalent inhibitor and has different HLA types, or providing the cell free peptide conjugate/MHC complex disclosed herein presented by two or more HLAs, and screening binding partners to thereby identify a binding partner that binds with specificity to the peptide conjugate or the antigen that is presented by more than one HLA. In some embodiments, the two or more HLAs types comprise HLA-A*02:01 and at least one additional HLA type. In some embodiments, the two or more HLAs are each HLA-A*02:01, HLA-A*03:01, or HLA- A*l l:01.
In another aspect, the present disclosure provides a method of killing a cancer cell in a subject, the method comprising administering to the subject: (a) a targeted covalent inhibitor, and (b) the binding partner disclosed herein, the polynucleotide disclosed herein, the vector disclosed herein, the pharmaceutical composition disclosed herein, or the cell disclosed herein.
In some embodiments, the targeted covalent inhibitor targets KRAS.
In another aspect, the present disclosure provides a method of targeting a cell that expresses KRASG12C in a subject that has been treated with a KRASG12C targeted covalent inhibitor, the method comprising administering to the subject a binding partner disclosed herein, the polynucleotide disclosed herein, the vector disclosed herein, the pharmaceutical composition disclosed herein, or the cell disclosed herein.
In some embodiments, the subject has cancer.
In another aspect, the present disclosure provides a method of treating cancer in a subject that has been treated with a targeted covalent inhibitor, the method comprising administering to the subject a binding partner disclosed herein, the polynucleotide disclosed herein, the vector disclosed herein, the pharmaceutical composition disclosed herein, or the cell disclosed herein. In another aspect, the present disclosure provides a method of treating a disease or disorder in a subject that has been treated with a targeted covalent inhibitor, the method comprising administering to the subject a binding partner disclosed herein, the polynucleotide disclosed herein, the vector disclosed herein, the pharmaceutical composition disclosed herein, or the cell disclosed herein. In some embodiments, the disease or disorder is an autoimmune disease.
In another aspect, the present disclosure provides a method of enhancing immune recognition of a cell expressing a KRASG12C mutation in a subject that has a cancer that exhibits a KRASG12C mutation, the method comprising administering to the subject: (a) a KRASG12C inhibitor, and a binding partner disclosed herein, the polynucleotide disclosed herein, the vector disclosed herein, the pharmaceutical composition disclosed herein, or the cell disclosed herein. In some embodiments, the subject has been previously treated with the KRASG12C inhibitor.
In some embodiments, the inhibitor is AMG-510 or MRTX849, JNJ74699157, LY3499446, MRTX-1257, JDQ443, MRTX-1133, or RMC 6291. In some embodiments, the inhibitor can be a beta-lactone (e.g., G12Si-l, G12Si-2, G12Si-3, G12Si-4 and G12Si-5).
In some embodiments, the targeted covalent inhibitor targets KRAS.
In some embodiments, the cancer is renal cell carcinoma, breast cancer, prostate cancer, pancreatic cancer, lung cancer, liver cancer, ovarian cancer, cervical cancer, colon cancer, esophageal cancer, glioma, glioblastoma, brain cancer, stomach cancer, bladder cancer, testicular cancer, head and neck cancer, melanoma, skin cancer, sarcoma, fibrosarcoma, angiosarcoma, osteosarcoma, rhabdomyosarcoma, leukemia, lymphoma, or myeloma.
In some embodiments, the targeted covalent inhibitor is sotorasib (e.g., AMG-510), adagrasib (e.g., MRTX849), divarasib (e.g., GDC6036), or any combination thereof
In some embodiments, the method disclosed herein further comprises administering an additional therapeutic agent. In some embodiments, the additional therapeutic agent is a chemotherapy, an immunomodulator, or another targeted covalent inhibitor. In some embodiments, the immunomodulator is a checkpoint targeting agent, optionally wherein the checkpoint targeting agent is selected from the group consisting of an antagonist anti-PD-1 antibody, an antagonist anti-PD-Ll antibody, an antagonist anti-PD-L2 antibody, an antagonist anti-CTLA-4 antibody, an antagonist anti-BTLA antibody, an antagonist anti- TREMR antibody, an antagonist anti-TIGIT antibody, an antagonist anti-VISTA antibody, an antagonist anti-TIM-3 antibody, an antagonist anti-LAG-3 antibody, an antagonist anti- CEACAM1 antibody, an agonist anti-GITR antibody, an agonist anti-OX40 antibody, and an agonist anti-CD137 antibody, an agonist anti-DR3 antibody, an agonist anti-TNFSF14 antibody, an agonist anti-CD27 antibody, an agonist anti-ICOS antibody, and an agonist anti- CD28 antibody; a cytokine, optionally wherein the cytokine is an anchored IL2 or an engineered IL2; an a2 adrenergic receptor agonist, or an inhibitor of extracellular adenosine (eADO) signaling. The cytokine can be an anchored cytokine (e.g., an anchored IL-2). The anchored cytokine can be a cytokine fused a collagen-binding protein. The cytokine can be an engineered cytokine. The engineered cytokine can be a variant or mutant of a wild-type cytokine.
In another aspect, the present disclosure provides a method of detecting a peptide conjugate/MHC complex in a biological sample, the method comprising contacting the sample with the binding partner disclosed herein, wherein the peptide conjugate/MHC complex comprises a peptide conjugate formed by the covalent reaction of a targeted covalent inhibitor with a peptide. In some embodiments, the peptide is an KRASG12C peptide. In some embodiments, the targeted covalent inhibitor is an KRASG12C inhibitor. In some embodiments, the targeted covalent inhibitor is sotorasib (e.g., AMG-510), adagrasib (e.g., MRTX849), divarasib (e.g., GDC6036), or any combination thereof. In some embodiments, the biological sample is blood or serum.
In another aspect, the present disclosure provides a method of identifying a cell containing the peptide conjugate/MHC complex disclosed herein, the method comprising contacting the cell with the binding partner disclosed herein.
In another aspect, the present disclosure provides a method for generating one or more binding partners that specifically bind to a peptide conjugate, wherein said peptide conjugate comprises a peptide covalently bound to a non-peptide molecule, the method comprising: (a) exposing said peptide conjugate to a plurality of binding partners, and b) selecting binding partners that specifically bind to said peptide conjugate to provide one or more selected binding partners. In some embodiments, the method disclosed herein further comprises selecting binding partners that specifically bind to said peptide conjugate but do not detectably bind, or bind with lower affinity, to said peptide or to said non-peptide molecule when they are not covalently bound to each other.
In another aspect, the present disclosure provides a method for generating one or more binding partners that specifically bind to a peptide conjugate presented in the context of a MHC molecule or a fragment or derivative thereof, wherein said peptide conjugate comprises a peptide covalently bound to a non-peptide molecule, the method comprising: (a) providing a complex of said peptide conjugate with said MHC molecule or the fragment or derivative thereof; (b) exposing said complex to a plurality of binding partners, and (c) selecting binding partners that specifically bind to said complex to provide one or more selected binding partners. In some embodiments, the method disclosed herein further comprises selecting binding partners that specifically bind to said complex but do not detectably bind or bind with lower affinity to a complex of said peptide with said MHC molecule or fragment thereof, wherein said peptide is not covalently bound to said non-peptide molecule. In some embodiments, the method disclosed herein further comprises selecting binding partners that specifically bind to said peptide conjugate presented in the context of two or more different MHC molecules.
In some embodiments, the MHC molecule is MHC class I molecule and the peptide is 7-15 amino acids long, or wherein the MHC molecule is a non-classical MHC class I molecule and the peptide is 7-15 amino acids long. In some embodiments, the MHC molecule is MHC class II molecule and the peptide is 9-30 amino acids long.
In some embodiments, the complex is immobilized on a solid support. In some embodiments, the complex is present on a surface of a cell.
In some embodiments, the method further comprises determining whether said one or more binding partners selected in step (c) can mediate immune cell-mediated killing or antibody-dependent cellular cytotoxicity (ADCC), antibody dependent cellular phagocytosis (ADCP) or complement dependent cytotoxicity (CDC)-mediated killing of said cell, or by use of said binding partners as immunotoxins or as antibody-drug conjugates (ADCs) or as radioconjugates. In some embodiments, the plurality of binding partners is generated by a phage-display library or yeast-display library.
In another aspect, the present disclosure provides a method of killing a cancer cell, the method comprising administering to said cell a binding partner which specifically binds to a peptide conjugate, wherein said peptide conjugate comprises (i) a peptide derived from a target protein within said cell which peptide is covalently bound to (ii) a non-peptide molecule, and wherein said binding partner mediates an immune cell-mediated killing or antibody-drug conjugate (ADC)-mediated or complement dependent cytotoxicity (CDC) immunotoxin killing of said cell, or by action of a radioconjugate, to thereby kill the cancer cell.
In some embodiments, the peptide conjugate is presented on said cell in the context of a MHC molecule. In some embodiments, the binding partner specifically binds to a complex comprising said peptide conjugate and said MHC molecule. In some embodiments, the binding partner does not detectably bind to a complex of said peptide with said MHC molecule, wherein said peptide is not covalently bound to said non-peptide molecule. In some embodiments, the method disclosed herein further comprises administering to said cell said non-peptide molecule, wherein said non-peptide molecule forms a covalent bond with said target protein in said cell. In some embodiments, the non-peptide molecule is administered to said cell prior to administering said binding partner. In some embodiments, the cell is isolated from a subject. In some embodiments, the cell is in a subject and said binding partner is administered to the subject. In some embodiments, the cell is in a subject and said non-peptide molecule and said binding partner are administered to the subject. In some embodiments, the target protein is alternatively spliced or over-expressed in cancer cells but is not alternatively spliced or over-expressed by cancer cells. In some embodiments, the target protein is encoded by a gene that is mutated in cancer cells but not mutated in non-cancer cells. In some embodiments, the non-peptide molecule is a covalent inhibitor of said target protein.
In some embodiments, binding partner is a component of a bi-specific T-cell engager (BiTE), bispecific killer cell engager (BiKE), CrossMab technology, or a chimeric antigen receptor (CAR), or comprises a radioconjugate. In some embodiments, the CAR is present on a T cell, natural killer (NK) cell, or killer macrophage. In some embodiments, the binding partner is an antibody-drug conjugate (ADC) or toxin conjugate.
In some embodiments, the target protein is selected from KRAS or a mutant thereof. In some embodiments, the target protein is a KRAS protein comprising a G12C mutation and said non-peptide molecule is selected from MRTX849, AMG-510, and derivatives thereof.
In some embodiments, the target protein is a KRAS protein comprising a G12C mutation and said non-peptide molecule is 2E07, 6H05, SML-8-73-1, MRTX849, JNJ74699157, LY3499446, ARS-853, ARS-1620, ARS-3284, GDC-6036, D-1553, JDQ443, BI 1823911, or derivatives thereof.
In some embodiments, the target protein is a KRAS protein comprising a G12S mutation and said non-peptide molecule can be a beta-lactone (e.g., G12Si-l, G12Si-2, G12Si- 3, G12Si-4 and G12Si-5).
In another aspect, the present disclosure provides a method of treating a cancer in a subject in need thereof, the method comprising administering to said subject an effective amount of a binding partner which specifically binds to a peptide conjugate, wherein said peptide conjugate comprises (i) a peptide derived from a target protein present in cancer cells of said subject, which peptide is covalently bound to (ii) a non-peptide molecule, and wherein said binding partner mediates immune cell-mediated killing or antibody-drug conjugates (ADC)-mediated killing of cancer cells, or complement dependent cytotoxicity (CDC) mediated killing of said cancer cell, or by use of said binding partners as immunotoxins in said subject, to thereby kill the cancer cell. In some embodiments, the peptide conjugate is presented on cancer cells of said subject in the context of an MHC molecule. In some embodiments, the binding partner specifically binds to a complex comprising said peptide conjugate and said MHC molecule. In some embodiments, the binding partner does not detectably bind to a complex of said peptide with said MHC molecule, wherein said peptide is not covalently bound to said non-peptide molecule. In some embodiments, the subject has previously received said non-peptide molecule. In some embodiments, the method disclosed herein further comprises administering to said subject an effective amount of said non-peptide molecule, wherein said non-peptide molecule forms a covalent bond with said target protein in cancer cells of said subject. In some embodiments, the non-peptide molecule is administered to said subject prior to administering said binding partner. In some embodiments, the non-peptide molecule is administered to said subject concomitantly to administering said binding partner.
In another aspect, the present disclosure provides a method for improving efficacy of an anti-cancer treatment in a subject in need thereof, wherein said anti-cancer treatment comprises administering to said subject a non-peptide molecule, which non-peptide molecule forms a covalent bond with a target protein in cancer cells of said subject, the method comprising further administering to said subject an effective amount of a binding partner which specifically binds to a peptide conjugate, wherein said peptide conjugate comprises a peptide derived from said target protein covalently bound to said non-peptide molecule, and wherein said binding partner mediates immune cell-mediated killing or antibody-drug conjugate (ADC)-mediated killing of cancer cells, or complement dependent cytotoxicity (CDC) mediated killing of said cancer cell, or by use of said binding partners as immunotoxins, or by use of said binding partners as radioconjugates, in said subject. In some embodiments, the peptide conjugate is presented on cancer cells of said subject in the context of an MHC molecule. In some embodiments, the binding partner specifically binds to a complex comprising said peptide conjugate and said MHC molecule. In some embodiments, the binding partner does not detectably bind to a complex of said peptide with said MHC molecule, wherein said peptide is not covalently bound to said non-peptide molecule. In some embodiments, the subject has previously received said non-peptide molecule prior to administering said binding partner.
In another aspect, the present disclosure provides a kit comprising (i) a non-peptide molecule, wherein said non-peptide molecule forms a covalent bond with a target protein in a cell, (ii) a binding partner that specifically binds to a peptide conjugate, wherein said peptide conjugate comprises a peptide derived from said target protein covalently bound to said nonpeptide molecule, and optionally (iii) instructions for use.
In another aspect, the present disclosure provides an isolated peptide conjugate comprising a peptide of 7-30 amino acids in length comprising an amino acid sequence that is at least 80% identical to the amino acid sequence VVGACGVGK, wherein the peptide is conjugated to AMG-510 or a derivative thereof.
In another aspect, the present disclosure provides an isolated molecular complex comprising the peptide conjugate disclosed herein and a MHC molecule, or a fragment or derivative thereof.
In another aspect, the present disclosure provides a host cell comprising the molecular complex disclosed herein.
In another aspect, the present disclosure provides a solid surface carrier comprising the molecular complex disclosed herein.
In another aspect, the present disclosure provides a kit comprising (i) the peptide conjugate disclosed herein, the molecular complex disclosed herein, the cell disclosed herein, the solid surface carrier of disclosed herein, or any combination thereof, and optionally (ii) instructions for use.
In another aspect, the present disclosure provides an isolated binding partner that specifically binds the peptide conjugate disclosed herein or the molecular complex disclosed herein. In some embodiments, the binding partner is an antibody or an antigen-binding fragment thereof. In some embodiments, the binding partner is a component of a chimeric antigen receptor (CAR).
In another aspect, the present disclosure provides a fusion protein comprising the binding partner disclosed herein, said fusion protein comprising at least one cytokine. In some embodiments, the cytokine is selected from the group consisting of interleukin (IL)-2, IL-7, IL- 8, IL- 15, IL- 17, IL- 18, IL-21, and a combination thereof. In some embodiments, the cytokine may be a modified (or mutated) cytokine (e.g., a modified interleukin (IL)-2, IL-7, IL-8, IL- 15, IL-17, IL-18, IL-21, or any combination thereof). In some embodiments, the fusion protein can comprise an agonist (e.g., an agonist to CD28 or 4-1BB). In some embodiments, the binding partner comprises a component that binds to a T cell or natural killer (NK) cell protein, wherein said T cell or NK cell protein is selected from the group consisting of a T cell receptor protein, CD4, CD8, CD28, CD 16 A, NKG2D, NKp30, NKp46, CD 137, and a combination thereof. In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain comprising a heavy chain variable region (VH) and/or a light chain variable region (VL). In some embodiments, the VH comprises a HC CDR3 comprising the amino acid sequence of LWASGLDY. In some embodiments, the VL comprises a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain comprising a heavy chain variable region (VH) and/or a light chain variable region (VL). In some embodiments, the VH comprises a HC CDR3 comprising the amino acid sequence of SYYGFWQALWALDY. In some embodiments, the VL comprises a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain comprising a heavy chain variable region (VH) and/or a light chain variable region (VL). In some embodiments, the VH comprises a HC CDR3 comprising the amino acid sequence of GYYYPYYAMDY. In some embodiments, the VL comprises a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain comprising a heavy chain variable region (VH) and/or a light chain variable region (VL). In some embodiments, the VH comprises a HC CDR3 comprising the amino acid sequence of GYYYPYYAMDY. In some embodiments, the VL comprises a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain comprising a heavy chain variable region (VH) and/or a light chain variable region (VL). In some embodiments, the VH comprises a HC CDR3 comprising the amino acid sequence of X1X2X3X4X5MDY. In some embodiments, the VL comprises a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain comprising a heavy chain variable region (VH) and/or a light chain variable region (VL). In some embodiments, the VH comprises a HC CDR3 comprising the amino acid sequence of SGYYSSHWYLQSWYQAMDY. In some embodiments, the VL comprises a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain comprising a heavy chain variable region (VH) and/or a light chain variable region (VL). In some embodiments, the VH comprises a HC CDR3 comprising the amino acid sequence of SGYYSSHWYLQSWYQAMDY. In some embodiments, the VL comprises a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain comprising a heavy chain variable region (VH) and/or a light chain variable region (VL). In some embodiments, the VH comprises a HC CDR3 comprising the amino acid sequence of HYSEKWWGWYTMYIDAMDY. In some embodiments, the VL comprises a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain comprising a heavy chain variable region (VH) and/or a light chain variable region (VL). In some embodiments, the VH comprises a HC CDR3 comprising the amino acid sequence of Xi WYYX2GMD Y. In some embodiments, the VL comprises a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain comprising a heavy chain variable region (VH) and/or a light chain variable region (VL). In some embodiments, the VH comprises a HC CDR3 comprising the amino acid sequence of Xi WYYX2GMD Y. In some embodiments, the VL comprises a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain comprising a heavy chain variable region (VH) and/or a light chain variable region (VL). In some embodiments, the VH comprises a HC CDR3 comprising the amino acid sequence of Xi WYYX2GMD Y. In some embodiments, the VL comprises a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain comprising a heavy chain variable region (VH) and/or a light chain variable region (VL). In some embodiments, the VH comprises a HC CDR3 comprising the amino acid sequence of SSRQYYHSQVEPPMAMDY. In some embodiments, the VL comprises a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain binds to a peptide conjugate/MHC complex, wherein the peptide conjugate of the peptide conjugate/MHC complex is a peptide covalently linked to a targeted covalent inhibitor or fragment thereof; and wherein the polypeptide binds to an epitope of the MHC.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain binds to a peptide conjugate/MHC complex, wherein the peptide conjugate of the peptide conjugate/MHC complex is a peptide covalently linked to a targeted covalent inhibitor or fragment thereof; and wherein the antigen-binding domain binds to the peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 50 nM.
In another aspect, the present disclosure provides a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain binds to a peptide conjugate/MHC complex, wherein the peptide conjugate of the peptide conjugate/MHC complex is a peptide covalently linked to a targeted covalent inhibitor or fragment thereof; and wherein the antigen-binding domain does not bind to the free targeted covalent inhibitor with a dissociation constant (KD) of less than 200 nM.
In an aspect, the present disclosure provides a polypeptide comprising an antigenbinding domain, wherein the antigen-binding domain binds to a first peptide conjugate/MHC complex and a second peptide conjugate/MHC complex, wherein the first peptide conjugate/MHC complex comprises a first peptide covalently linked to a first targeted covalent inhibitor or fragment thereof (e.g., AMG-510) and a first MHC, and the second peptide conjugate/MHC complex comprises a second peptide covalently linked to a second targeted covalent inhibitor or fragment thereof (e.g., MRTX849) and a second MHC. In an aspect, the polypeptide binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 100 nM. In some embodiments, the polypeptide binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 75 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 50 nM. In some embodiments, the antigenbinding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 40 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 30 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 20 nM. In some embodiments, the antigenbinding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 10 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 5 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 1 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 0.1 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 10 pM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 5 pM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 1 pM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 10 nM, at most about 9 nM, at most about 8 nM, at most about 7 nM, at most about 6 nM, at most about 5 nM, at most about 4 nM, at most about 3 nM, at most about 2 nM, at most about 1 nM, at most about 1 nM, at most about 0.9 nM, at most about 0.8 nM, at most about 0.7 nM, at most about 0.6 nM, at most about 0.5 nM, at most about 0.4 nM, at most about 0.3 nM, at most about 0.2 nM, or at most about 0.1 nM.
In an aspect, the polypeptide binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 100 nM. In some embodiments, the polypeptide binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 75 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 50 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 40 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 30 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 20 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 10 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 5 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 1 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 0.1 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 10 pM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 5 pM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 1 pM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of equal to or less than about 10 nM, equal to or less than about 9 nM, equal to or less than about 8 nM, equal to or less than about 7 nM, equal to or less than about 6 nM, equal to or less than about 5 nM, equal to or less than about 4 nM, equal to or less than about 3 nM, equal to or less than about 2 nM, equal to or less than about 1 nM, equal to or less than about 1 nM, equal to or less than about 0.9 nM, equal to or less than about 0.8 nM, equal to or less than about 0.7 nM, equal to or less than about 0.6 nM, equal to or less than about 0.5 nM, equal to or less than about 0.4 nM, equal to or less than about 0.3 nM, equal to or less than about 0.2 nM, or equal to or less than about 0.1 nM.
In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor (e.g., AMG-510 or MRTX849) with a dissociation constant (KD) of less than 150 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 200 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 250 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 300 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 350 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 400 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 450 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 500 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 1 gM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 5 gM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 10 gM.
In some embodiments, the antigen-binding domain does not bind to a free second target covalent inhibitor (e.g., AMG-510 or MRTX849) with a dissociation constant (KD) of less than 150 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 200 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 250 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 300 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 350 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 400 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 450 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 500 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 1 gM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 5 gM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 10 gM. In some embodiments, the antigen-binding domain does not bind to a free first peptide with a dissociation constant (KD) of less than 150 nM. In some embodiments, the antigenbinding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 200 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 250 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 300 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 350 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 400 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 450 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 500 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 1 pM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 5 pM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 10 pM.
In some embodiments, the antigen-binding domain does not bind to a free second peptide with a dissociation constant (KD) of less than 150 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 200 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 250 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 300 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 350 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 400 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 450 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 500 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 1 pM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 5 pM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 10 pM.
In some embodiments, the antigen-binding domain does not bind to a free first peptide conjugate (e.g., a peptide conjugate not complexed with an MHC molecule) with a dissociation constant (KD) of less than 150 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 200 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 250 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 300 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 350 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 400 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 450 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 500 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 1 pM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 5 pM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 10 pM.
In some embodiments, the antigen-binding domain does not bind to a free second peptide conjugate (e.g., a peptide conjugate not complexed with an MHC molecule) with a dissociation constant (KD) of less than 150 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 200 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 250 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 300 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 350 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 400 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 450 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 500 nM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 1 pM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 5 pM. In some embodiments, the antigen-binding domain does not bind to a free first target covalent inhibitor with a dissociation constant (KD) of less than 10 pM.
In some embodiments, the antigen-binding domain does not bind to: (i) a free targeted covalent inhibitor (e.g., AMG-510 and/or MRTX849) with a dissociation constant (KD) of less than 150 nM, less than 200 nM, less than 250 nM, less than 300 nM, less than 350 nM, less than 400 nM, less than 450 nM, less than 500 nM, less than 1 pM, less than 5 pM, or less than 10 pM; (ii) a free peptide with a dissociation constant (KD) of less than 150 nM, less than 200 nM, less than 250 nM, less than 300 nM, less than 350 nM, less than 400 nM, less than 450 nM, less than 500 nM, less than 1 pM, less than 5 pM, or less than 10 pM; (iii) a free peptide conjugate (e.g., a peptide conjugate no complexed with an MHC molecule) with a dissociation constant (KD) of less than 150 nM, less than 200 nM, less than 250 nM, less than 300 nM, less than 350 nM, less than 400 nM, less than 450 nM, less than 500 nM, less than 1 pM, less than 5 pM, or less than 10 pM; or any combination of the above.
In an aspect, the present disclosure provides a polypeptide comprising an antigenbinding domain, wherein the antigen-binding domain binds to a first peptide conjugate/MHC complex and a second peptide conjugate/MHC complex, wherein the first peptide conjugate/MHC complex comprises a first peptide covalently linked to a first targeted covalent inhibitor or fragment thereof (e.g., AMG-510) and a first MHC, and the second peptide conjugate/MHC complex comprises a second peptide covalently linked to a second targeted covalent inhibitor or fragment thereof (e.g., MRTX849) and a second MHC. In an aspect, the polypeptide binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 100 nM. In some embodiments, the polypeptide binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 75 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 50 nM. In some embodiments, the antigenbinding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 40 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 30 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 20 nM. In some embodiments, the antigenbinding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 10 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 5 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 1 nM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 0.1 nM. In some embodiments, the antigen-binding domain binds to the peptide conjugate/MHC disclosed herein complex with a dissociation constant (KD) of at most about 50 nM, at most about 40 nM, at most about 30 nM, at most about 20 nM, at most about 10 nM, at most about 1 nM, at most about 0.1 nM, at most about 10 pM, at most about 1 pM, or at most about 0.1 pM. In some embodiments, the antigen-binding domain binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 10 nM, at most about 9 nM, at most about 8 nM, at most about 7 nM, at most about 6 nM, at most about 5 nM, at most about 4 nM, at most about 3 nM, at most about 2 nM, at most about 1 nM, at most about 1 nM, at most about 0.9 nM, at most about 0.8 nM, at most about 0.7 nM, at most about 0.6 nM, at most about 0.5 nM, at most about 0.4 nM, at most about 0.3 nM, at most about 0.2 nM, or at most about 0.1 nM.
In an embodiment, the affinity of the binding partner for the peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the binding partner for the free peptide or free targeted covalent inhibitor. In an embodiment, the affinity of the binding partner for the peptide conjugate/MHC complex is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, at least 1,000 times, at least 2,500 times, at least 5,000 times, at least 10,000, at least 20,000, at least 30,000, at least 40,000, or at least 50,000 times greater than the affinity of the binding partner to the free targeted covalent inhibitor or the free peptide conjugate. In some embodiments, the free targeted covalent inhibitor comprises AMG-510 or MRTX849. In some embodiments, the affinity of the binding partner to the free drug is reported as ICso. In some embodiments, the affinity of the binding partner to the free drug is reported as an ICso of more than 1 pM. In some embodiments, the affinity of the binding partner to the free drug is reported as an ICso of more than 5 pM. In some embodiments, the affinity of the binding partner to the free drug is reported as an ICso of more than 10 pM. In some embodiments, the affinity of the binding partner to the free drug is reported as an ICso of more than 15 pM. In some embodiments, the affinity of the binding partner to the free drug is reported as an ICso of more than 20 pM. In some embodiments, the affinity of the binding partner to the free drug is reported as an ICso of more than 30 pM. In some embodiments, the affinity of the binding partner to the free drug is reported as an ICso of more than 40 pM. In some embodiments, the affinity of the binding partner to the free drug is reported as an ICso of more than 50 pM. In some embodiments, the affinity of the binding partner to the free drug is reported as an ICso of more than 75 pM. In some embodiments, the affinity of the binding partner to the free drug is reported as an ICso of more than 100 pM. In some embodiments, the affinity of the binding partner to the free drug is reported as an ICso of more than 200 pM.
In some embodiments, the antigen-binding domain does not detectably bind to the free targeted covalent inhibitor or binds with lower affinity to the free targeted covalent inhibitor compared to binding affinity to the peptide conjugate/MHC complex. In some embodiments, the targeted covalent inhibitor is sotorasib (e.g., AMG-510), adagrasib (e.g., MRTX849), divarasib (e.g., GDC6036), or any combination thereof. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with an ICso of at least 50 nM. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with an ICso of at least 75 nM. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with an ICso of at least 100 nM. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with an ICso of at least 150 nM. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with an IC50 of at least 200 nM. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with an IC50 of at least 250 nM. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with an IC50 of at least 300 nM. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with an IC50 of at least 3500 nM. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with an IC50 of at least 400 nM. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with an IC50 of at least 450 nM. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with an IC50 of at least 500 nM. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with an IC50 of at least 750 nM. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with an IC50 of at least 1000 nM.
The binding partner described herein may not bind to the peptide/MHC complex without the conjugate (e.g., the targeted covalent inhibitor). In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) from 10 pM to 50 nM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) from 1 pM to 10 pM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) from 0.01 pM to 10 pM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) from 0.1 nM to 1 nM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) from 1 nM to 2 nM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) from 2 nM to 5 nM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) of equal to less than about 5 nM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) of equal to less than about 4 nM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) of equal to less than about 3 nM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) of equal to less than about 2 nM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) of equal to less than about 1 nM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) of equal to less than about 0.1 nM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) of about equal to less than 0.01 nM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) of equal to less than about 1 pM. In some cases, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as a dissociation constant (KD) of equal to less than about 0.1 pM.
In some embodiments, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as ICso or ECso. In some embodiments, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as an ICso of less than about 1 pM. In some embodiments, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as an ICso of less than about 10 pM. In some embodiments, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as an ICso of less than about 100 pM. In some embodiments, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as an ICso of less than about 1000 pM. In some embodiments, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as an ICso of less than about 1 nM. In some embodiments, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as an ICso of less than about 10 nM. In some embodiments, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as an ICso of less than about 100 nM. In some embodiments, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as an ICso of less than about 1000 nM. In some embodiments, the affinity of the binding partner to the peptide conjugate/MHC complex is reported as an ICso of less than about 1 pM.
In some embodiments, the antibody or the antigen-binding fragment specifically binds to a peptide conjugate/MHC complex and wherein the antibody or the antigen-binding fragment interacts with the MHC of the peptide conjugate/MHC complex. In some embodiments, the peptide conjugate/MHC complex comprises: (a) a peptide conjugate comprising a peptide covalently linked to a targeted covalent inhibitor or a fragment thereof; and (b) an MHC. In some embodiments, the MHC is a human leukocyte antigen (HLA). In some embodiments, the HLA is HLA-A*03:01, HLA-A* 11 :01, and/or HL A-A *02:01.
In some embodiments, the antigen-binding domain binds to the peptide conjugate/MHC complex with a greater affinity than to the peptide or free targeted covalent inhibitor. In some embodiments, the antigen-binding domain binds to the peptide conjugate/MHC complex disclosed herein with a dissociation constant (KD) of at most about 50 nM, at most about 40 nM, at most about 30 nM, at most about 20 nM, at most about 10 nM, at most about 1 nM, at most about 0.1 nM, at most about 10 pM, at most about 1 pM, or at most about 0.1 pM. In some embodiments, the antigen-binding domain binds to the peptide conjugate/MHC complex with a dissociation constant (KD) of from about 0.01 nM to about 20 nM.
The antigen-binding domain described herein may not bind to a free targeted covalent inhibitor. The antigen-binding domain described herein may not detectably bind to a free targeted covalent inhibitor. For example, the antigen-binding domain described herein may bind to the free targeted covalent inhibitor with at least 10 times, 100 times, 1,000 times, 10,000 times, 100,000 times or more greater dissociation constant (KD) than the KD of the antigen-binding domain binding to the peptide conjugate/MHC complex. In some embodiments, the antigen-binding domain binds to the free targeted covalent inhibitor with a dissociation constant (KD) of at least about 150 nM, at least about 200 nM, at least about 300 nM, at least about 400 nM, at least about 500 nM, at least about 1 pM, at least about 10 pM, at least about 20 pM, at least about 30 pM, at least about 40 pM, at least about 50 pM, or at least 100 pM. In some embodiments, the antigen -binding domain binds to the free targeted covalent inhibitor with a dissociation constant (KD) of at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, at least 1,000 times, at least 2,500 times, at least 5,000 times, at least 10,000, at least 20,000, at least 30,000, at least 40,000, or at least 50,000 times greater than a KD of the antibody or the antigen-binding fragment binding to the peptide conjugate/MHC complex.
The antigen-binding domain described herein may not bind to a free peptide conjugate that is not in complex with an MHC molecule. The antigen-binding domain described herein may not detectably bind to a free peptide conjugate. For example, the antigen-binding domain described herein may bind to the free peptide conjugate with at least 10 times, 100 times, 1,000 times, 10,000 times, 100,000 times or more greater dissociation constant (KD) than the KD of the antigen-binding domain binding to the peptide conjugate/MHC complex. In some embodiments, the antigen-binding domain binds to the free peptide conjugate with a dissociation constant (KD) of more than about 150 nM, more than about 200 nM, more than about 300 nM, more than about 400 nM, more than about 500 nM, more than 1 pM, more than 10 pM, more than 20 pM, more than 30 pM, more than 40 pM, more than 50 pM, or more than 100 pM. In some embodiments, the antigen-binding domain binds to the free peptide conjugate with a dissociation constant (KD) that is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, at least 1,000 times, at least 2,500 times, at least 5,000 times, at least 10,000, at least 20,000, at least 30,000, at least 40,000, or at least 50,000 times greater than a KD of the antibody or the antigen-binding fragment to the peptide conjugate/MHC complex. In some embodiments, the antigen-binding domain binds to the free peptide with a dissociation constant (KD) of more than about 150 nM, more than about 200 nM, more than about 300 nM, more than about 400 nM, more than about 500 nM, more than 1 pM, more than 10 pM, more than 20 pM, more than 30 pM, more than 40 pM, more than 50 pM, or more than 100 pM. In some embodiments, the antigen-binding domain binds to the free peptide with a dissociation constant (KD) that is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, at least 1,000 times, at least 2,500 times, at least 5,000 times, at least 10,000, at least 20,000, at least 30,000, at least 40,000, or at least 50,000 times greater than a KD of the antibody or the antigen-binding fragment to the peptide conjugate/MHC complex. In some embodiments, the antigen-binding domain binds to the peptide conjugate/MHC complex with an affinity that is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, at least 1,000 times, at least 2,500 times, at least 5,000 times, or at least 10,000 times greater than the affinity of the antibody or the antigen-binding fragment to the free targeted covalent inhibitor or the free peptide conjugate. In some embodiments, the binding partner does not detectably bind to a complex of a peptide with a MHC molecule, wherein the peptide is not covalently bound to a non-peptide molecule. In some embodiments, the binding partner binds at less than lOx affinity to a complex of a peptide with a MHC molecule, wherein the peptide is not covalently bound to a non-peptide molecule. In some embodiments, the binding partner binds at less than lOOx affinity to a complex of a peptide with a MHC molecule, wherein the peptide is not covalently bound to a non-peptide molecule. In some embodiments, the binding partner binds at less than l,000x affinity to a complex of a peptide with a MHC molecule, wherein the peptide is not covalently bound to a non-peptide molecule.
In some embodiments, the binding partner can be a polypeptide. In some embodiments, the polypeptide can have an antigen-binding domain. The antigen-binding domain can bind to the peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 50 nM, at most about 40 nM, at most about 30 nM, at most about 20 nM, at most about 10 nM, at most about 1 nM, at most about 0.1 nM, at most about 10 pM, at most about 1 pM, or at most about 0.1 pM. The antigen-binding domain can bind to the peptide conjugate/MHC complex with a dissociation constant (KD) from about 0.01 nM to about 20 nM. In some embodiments, the antigen-binding domain can bind to the free targeted covalent inhibitor with a dissociation constant (KD) of at least about 150 nM, at least about 200 nM, at least about 300 nM, at least about 400 nM, at least about 500 nM, at least about 1 pM, at least about 10 pM, at least about 20 pM, at least about 30 pM, at least about 40 pM, at least about 50 pM, or at least 100 pM. The antigen-binding domain can bind to the free targeted covalent inhibitor with a dissociation constant (KD) of at least about 10, at least about 100, at least about 1,000, at least about 2,500, at least about 5,000, at least about 10,000, at least about 20,000, at least about 30,000, at least about 40,000, at least about 50,000, at least about 60,000, at least about 70,000, at least about 80,000, at least about 90,000, or at least about 100,000 times more than a KD of the antigen-binding domain binding to the peptide conjugate/MHC complex. The antigen-binding domain can bind to the free peptide conjugate with a dissociation constant (KD) of more than about 100 nM, more than about 200 nM, more than about 300 nM, more than about 400 nM, more than about 500 nM, more than 1 pM, more than 10 pM, more than 20 pM, more than 30 pM, more than 40 pM, more than 50 pM, or more than 100 pM. The antigen-binding domain can bind to the free peptide conjugate with a dissociation constant (KD) at least about 10, at least about 100, at least about 1,000, at least about 2,500, at least about 5,000, at least about 10,000, at least about 20,000, at least about 30,000, at least about 40,000, at least about 50,000, at least about 60,000, at least about 70,000, at least about 80,000, at least about 90,000, or at least about 100,000 times more than a KD of the antigen-binding domain binding to the peptide conjugate/MHC complex. In some embodiments, the antigen-binding domain can bind to the free peptide with a dissociation constant (KD) of at least about 150 nM, at least about 200 nM, at least about 300 nM, at least about 400 nM, at least about 500 nM, at least about 1 pM, at least about 10 pM, at least about 20 pM, at least about 30 pM, at least about 40 pM, at least about 50 pM, or at least 100 pM. The antigen-binding domain can bind to the free peptide with a dissociation constant (KD) of at least about 10, at least about 100, at least about 1,000, at least about 2,500, at least about 5,000, at least about 10,000, at least about 20,000, at least about 30,000, at least about 40,000, at least about 50,000, at least about 60,000, at least about 70,000, at least about 80,000, at least about 90,000, or at least about 100,000 times more than a KD of the antigen-binding domain binding to the peptide conjugate/MHC complex. The antigen-binding domain can bind to the peptide conjugate/MHC complex with an affinity that is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, at least 1,000 times, at least 2,500 times, at least 5,000 times, or at least 10,000 times greater than the affinity of antigen-binding domain to the free targeted covalent inhibitor, the free peptide conjugate, or the free peptide.
In an embodiment, the MHC is a human leukocyte antigen (HLA). In an embodiment, the HLA is any HLA found in the Allele Frequency Net Database (allelefirequencies.net). In an embodiment, the HLA is HLA-A*02:01, HLA-A*03:01, and/or HLA-A* 11 :01.
In an aspect, provided herein is a cell-free peptide conjugate/MHC complex comprising: (a) a peptide conjugate formed by the covalent reaction of a targeted covalent inhibitor with a peptide; and (b) an MHC. In an embodiment, the peptide comprises a segment of KRASG12C. In some embodiments, the targeted covalent inhibitor is sotorasib (e.g., AMG-510), adagrasib e.g., MRTX849), divarasib (e.g., GDC6036), or any combination thereof. In some embodiments, the targeted covalent inhibitor is 2E07, 6H05, SML-8-73-1, MRTX849, JNJ74699157, LY3499446, ARS-853, ARS-1620, ARS-3284, GDC-6036, D-1553, JDQ443, BI 1823911, or derivatives thereof.
In an aspect, provided herein is a cell free peptide conjugate/MHC complex comprising: (a) a compound selected from the group consisting of compounds 1 and 2,
(2); covalently bonded to a cysteine residue in a peptide comprising the amino acid sequence of VVVGACGVGK, VVGACGVGK or KLVVVGACGV; and (b) an MHC. In an embodiment the MHC is an HLA, optionally wherein the HLA is HLA- A* 02 : 01 , HL A- A* 03 : 01 , or HL A- A* 11 :01.
In an aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of AMG-510 (e.g., sotorasib) with a KRASG12C peptide presented by an HLA, MRTX849 (e.g., adagrasib) with a KRASG12C peptide presented by an HLA, or GDC6036 (e.g., divarasib) with a KRASG12C peptide presented by an HLA, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of LWASGLDY; and/or (b) the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In an aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of AMG-510 (e.g, sotorasib) with a KRASG12C peptide presented by an HLA, MRTX849 (e.g, adagrasib) with a KRASG12C peptide presented by an HLA, or GDC6036 (e.g., divarasib) with a KRASG12C peptide presented by an HLA, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of SYYGFWQALWALDY; and/or (b) the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4is G, L, S, E or Y, Xs is W S, Y, E, or F, Xe is P, W, G, E, or Q, and X7 is I, F, or V. In an aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of AMG-510 (e.g., sotorasib) with a KRASG12C peptide presented by an HLA MRTX849 (e.g., adagrasib) with a KRASG12C peptide presented by an HL A, or GDC6036 (e.g., divarasib) with a KRASG12C peptide presented by an HLA, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of GYYYPYYAMDY; and/or (b) the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In an aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of AMG-510 (e.g, sotorasib) with a KRASG12C peptide presented by an HLA, MRTX849 (e.g, adagrasib) with a KRASG12C peptide presented by an HLA, or GDC6036 (e.g., divarasib) with a KRASG12C peptide presented by an HLA, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of GYYYPYYAMDY; and/or (b) the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In an aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of AMG-510 (e.g., sotorasib) with a KRASG12C peptide presented by an HLA, MRTX849 (e.g., adagrasib) with a KRASG12C peptide presented by an HL A, or GDC6036 (e.g., divarasib) with a KRASG12C peptide presented by an HL A, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of X1X2X3X4X5MDY, wherein Xi is F or G, X2 is Q or Y, X3 is W or G, X4 is Y or W, and X5 is A or G; and/or (b) the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In an aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of AMG-510 (e.g., sotorasib) with a KRASG12C peptide presented by an HL A, MRTX849 (e.g., adagrasib) with a KRASG12C peptide presented by an HL A, or GDC6036 (e.g., divarasib) with a KRASG12C peptide presented by an HL A, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of SGYYSSHWYLQSWYQAMDY; and/or (b) the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In an aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of AMG-510 (e.g., sotorasib) with a KRASG12C peptide presented by an HL A, MRTX849 (e.g., adagrasib) with a KRASG12C peptide presented by an HL A, or GDC6036 (e.g., divarasib) with a KRASG12C peptide presented by an HL A, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of SGYYSSHWYLQSWYQAMDY; and/or (b) the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
In an aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of AMG-510 (e.g., sotorasib) with a KRASG12C peptide presented by an HL A, MRTX849 (e.g., adagrasib) with a KRASG12C peptide presented by an HL A, or GDC6036 (e.g., divarasib) with a KRASG12C peptide presented by an HL A, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of HYSEKWWGWYTMYIDAMDY; and/or (b) the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4is G, L, S, E or Y, Xs is W S, Y, E, or F, Xe is P, W, G, E, or Q, and X7 is I, F, or V.
In an aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of AMG-510 (e.g, sotorasib) with a KRASG12C peptide presented by an HL A, MRTX849 (e.g, adagrasib) with a KRASG12C peptide presented by an HL A, or GDC6036 (e.g., divarasib) with a KRASG12C peptide presented by an HL A, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L; and/or (b) the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or a LC CDR3 comprising the amino acid comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In an aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of AMG-510 (e.g., sotorasib) with a KRASG12C peptide presented by an HL A, MRTX849 (e.g., adagrasib) with a KRASG12C peptide presented by an HL A, or GDC6036 (e.g., divarasib) with a KRASG12C peptide presented by an HL A, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L; and/or the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
In an aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of AMG-510 (e.g, sotorasib) with a KRASG12C peptide presented by an HL A, MRTX849 (e.g, adagrasib) with a KRASG12C peptide presented by an HL A, or GDC6036 (e.g., divarasib) with a KRASG12C peptide presented by an HL A, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of Xi WYYX2GMD Y, wherein Xi is G or Y and X2 is M or L; and/or (b) the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
In an aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of AMG-510 (e.g., sotorasib) with a KRASG12C peptide presented by an HL A, MRTX849 (e.g., adagrasib) with a KRASG12C peptide presented by an HL A, or GDC6036 (e.g., divarasib) with a KRASG12C peptide presented by an HL A, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of SSRQYYHSQVEPPMAMDY; and/or (b) the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or a LC CDR3 comprising the amino acid comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
In some embodiments, a polypeptide can comprise an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein: (a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of FX2X3X4AMDY, wherein X2 is Y, L, T, H, E, A, or R; X3 is D, V, L, E, H, T or R; and X4 is L or Y (SEQ ID NO: 267); and/or (b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX3SWLX7WX9X10T, wherein X3 is A or S, X7 is Y or H, X9 is L, K, V, Y or I, and X10 is L, V, or I (SEQ ID NO: 268).
In an aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of AMG-510 (e.g., sotorasib) with a KRASG12C peptide presented by an HL A, MRTX849 (e.g., adagrasib) with a KRASG12C peptide presented by an HL A, or GDC6036 (e.g., divarasib) with a KRASG12C peptide presented by an HL A, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; and/or a HC CDR3 comprising an amino acid sequence of FX2X3X4AMDY, wherein X2 is Y, L, T, H, E, A, or R; X3 is D, V, L, E, H, T or R; and XHs L or Y (SEQ ID NO: 267); and/or (b) the VL comprises: a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or a LC CDR3 comprising the amino acid comprising the amino acid sequence of QQX3SWLX7WX9X10T, wherein X3 is A or S, X7 is Y or H, X9 is L, K, V, Y or I, and X10 is L, V, or I (SEQ ID NO: 268). In some embodiments, the VH comprises: a HC CDR3 comprising the amino acid sequence of FX2X3X4AMDY, wherein X2 is Q, Y, L, T, H, E, A, or R; X3 is W, D, V, L, E, H, T or R; and X4 is L or Y.
In some embodiments, the VH comprises: (i) HC CDR1 comprising an amino acid sequence of X1SX3X4SIH, wherein Xi is I, F or V, X3 is S or Y, and X4 is S or Y; (ii) a HC CDR2 comprising an amino acid sequence of X1ISX4X5X6X7X8TX10YADSVKG, wherein Xi is S or Y, X4 is S or P, X5 is S or Y, Xe is S or Y, X7 is S or G, Xs is S or Y, and X10 is S or Y, and (iii) a HC CDR3 comprising an amino acid sequence of XNX1X2DY, wherein XN is an amino acid sequence selected from LWAS, FQWY, GYGW, GWYYL, YWYYM, YWYYL, GYYYPYY, SYYGFWQALW, SSRQYYHSQVEPPM, SGYYSSHWYLQSWYQ, and HYSEKWWGWYTMYID, Xi is G or A, and X2 is L or M. In some embodiments, the VL comprises (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and (iii) a LC CDR3 comprising the amino acid sequence of QQX3X4X5X6X7X8X9X10T, wherein X3 is W, T, S, A or G, X4 is N, W, S, G, Y, D or K, X5 is W, Y, S, A or T, X6 is G, S, Y, L, W, E or D, X7 is W, S, H, Y, E, F or absent, X8 is P, Q, S, W, E, L, G or absent, X9 is L or P, and X10 is I, L, F, or V.
In an aspect, described herein is a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL). In some embodiments, the VH comprises a HC CDR3 comprising an amino acid sequence of XNX1X2DY, wherein XN IS an amino acid sequence selected from LWAS, FQWY, GYGW, GWYYL, YWYYM, YWYYL, GYYYPYY, SYYGFWQALW, SSRQYYHSQVEPPM, SGYYSSHWYLQSWYQ, and HYSEKWWGWYTMYID, Xi is G or A, and X2 is L or M. In some embodiments, the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising at least 1, 2, 3, 4, 5, 6, 7, or 8 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and (iii) a LC CDR3 comprising the amino acid sequence of QQX3X4X5X6X7X8X9X10T, wherein X3 is W, T, S, A or G, X4 is N, W, S, G, Y, D or K, X5 is W, Y, S, A or T, X6 is G, S, Y, L, W, E or D, X7 is W, S, H, Y, E, F or absent, X8 is P, Q, S, W, E, L, G or absent, X9 is L or P, and X10 is I, L, F, or V. In some embodiments, the VH further comprises a HC CDR1 comprising an amino acid sequence of X1SX3X4SIH, wherein Xi is I, F or V, X3 is S or Y, and X4 is S or Y. In some embodiments, the VH further comprises a HC CDR2 comprising an amino acid sequence of X1ISX4X5X6X7X8TX10YADSVKG, wherein Xi is S or Y, X4 is S or P, X5 is S or Y, X6 is S or Y, X7 is S or G, Xs is S or Y, and X10 is S or Y. In some embodiments, the VL comprises a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes. In some embodiments, the VL comprises a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
In an aspect, described herein is a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of X1SX3X4SIH, wherein Xi is I, F or V, X3 is S or Y, and X4is S or Y; (ii) a HC CDR2 comprising an amino acid sequence of X1ISX4X5X6X7X8TX10YADSVKG, wherein Xi is S or Y, X4 is S or P, X5 is S or Y, Xe is S or Y, X7 is S or G, Xs is S or Y, and Xio is S or Y, and (iii) a HC CDR3 comprising an amino acid sequence of XNX1X2DY, wherein XN IS an amino acid sequence selected from LWAS, FQWY, GYGW, GWYYL, YWYYM, YWYYL, GYYYPYY, SYYGFWQALW, SSRQYYHSQVEPPM, SGYYSSHWYLQSWYQ, and HYSEKWWGWYTMYID, Xi is G or A, and X2 is L or M; and/or the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and a LC CDR3 comprising the amino acid sequence of QQX3X4X5X6X7X8X9X10T, wherein X3 is W, T, S, A or G, X4 is N, W, S, G, Y, D or K, X5 is W, Y, S, A or T, X6 is G, S, Y, L, W, E or D, X7 is W, S, H, Y, E, F or absent, X8 is P, Q, S, W, E, L, G or absent, X9 is L or P, and X10 is I, L, F, or V.
The CDR sequences, VL sequences, and VH sequences of exemplary antibodies that bind a conjugate formed by the covalent reaction of AMG-510 with a KRAS(G12C) peptide, MARTX849 with a KRAS(G12C) peptide, and/or GDC6036 with a KRAS(G12C) peptide presented on HLA-A*03:01, HLA-A*11 :01, and HLA-A*02:01 are provided in Table B below.
Table B: Sequences of exemplary antibodies that bind a conjugate formed by the covalent reaction of sotorasib (e.g., AMG-510), adagrasib (e.g., MRTX849), or divarasib (e.g., GDC6036) and a peptide presented on HLA-A*03:01, HLA-A*ll:01, and HLA-A*02:01.
In an embodiment, the binding partner comprises the HC CDR1, HC CDR2, and HC CDR3 of a heavy chain variable region amino acid sequence and/or the LC CDR1, LC CDR2, and LC CDR3 of a light chain variable region of a binding partner selected from the group consisting of RM_001, RM_002, RM_003, RM_004, RM_005, RM_006, RM_007, RM_008, RM_009, RM_010, RM_011, RM_012, RM_013, RM_014, RM_015, RM_016, RM_017, RM_018, RM_019, RM_020, RM_021, RM_022, RM_023, RM_024, RM_025, RM_026, RM_027, RM_028, RM_029, RM_030, RM_031, RM_032 (Tables C and D) , or a variant thereof comprising 1-5 amino acid changes in one or more of the CDR amino acid sequences.
In an embodiment, the binding partner can comprise a VH and/or a VL amino acid sequence having at least about 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% sequence identity to the following VH and VL sequences described herein.
In an embodiment, the binding partner can comprise a VH and/or a VL amino acid sequence having at least about 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% sequence identity to a VH and/or a VL of RM_001, RM_002, RM_003, RM_004, RM_005, RM_006, RM_007, RM_008, RM_009, RM_010, RM_011, RM_012, RM_013, RM_014, RM_015, RM_016, RM_017, RM_018, RM_019, RM_020, RM_021, RM_022, RM 023, RM_024, RM_025, RM_026, RM_027, RM_028, RM_029, RM_030, RM_031, RM_032 (Tables C and D).
A non-limiting example of a reference binding partner that binds to a conjugate formed by the covalent reaction of MRTX849 with a KRASG12C peptide and presented by an HLA molecule is referred to herein as RM 010.
Table C: Heavy chain CDR sequences of exemplary antibodies that bind a conjugate formed by the covalent reaction of AMG-510, MRTX849, or GDC6036 with a KRAS(G12C) peptide presented on HLA-A*03:01, HLA-A*ll:01, and HLA-A*02:01.
Table D:_Light chain CDR sequences of exemplary antibodies that bind a conjugate formed by the covalent reaction of AMG-510 or MRTX849 with a KRAS(G12C) peptide presented on HLA-A*03:01, HLA-A*ll:01, and HLA-A*02:01.
In an aspect, provided herein is a binding partner (e.g., a polypeptide with an antigenbinding domain or fragment thereof) that has superior binding affinity to a peptide- conjugate/MHC complex compared to the binding affinity to a free peptide, a free targeted covalent inhibitor (e.g., a free drug), a peptide conjugate, or a peptide/MHC complex. In some embodiments, the targeted covalent inhibitor of the present disclosure is sotorasib (e.g., AMG-510), adagrasib (e.g., MRTX849), divarasib (e.g., GDC6036), or any combination thereof. In some embodiments, the binding partner binds with similar affinity (with a KD value between about 1 nM and about 25 nM) to a peptide conjugate/MHC complex comprising AMG-510, MRTX849, or GDC6036. The peptide of the present disclosure can be a KRAS peptide. The KRAS peptide can have a mutation, e.g., KRASG12C, KRASG12D, KRASG12S, or KRASG12R. In some embodiments, the peptide conjugate of the present disclosure comprises a KRASG12C peptide covalently linked to AMG-510, MRTX849, or GDC6036. In some embodiments, the peptide conjugate is formed by a covalent reaction between the targeted covalent inhibitor and a electrophilic or nucleophilic residue of the peptide. The electrophilic or nucleophilic residue can be a cysteine residue, an aspartic acid residue, an arginine residue, a serine residue, or a tyrosine residue. In some embodiments, the peptide conjugate is formed by a covalent reaction between AMG-510 and a cysteine residue in KRASG12C. In some embodiments, the peptide conjugate is formed by a covalent reaction between MRTX849 and a cysteine residue in KRASG12C. In some embodiments, the peptide conjugate is formed by a covalent reaction between GDC6036 and a cysteine residue in KRASG12C. In some embodiments, the peptide conjugate (e.g., KRAS(G12C)-AMG-510, KRAS(G12C)-MRTX849, or KRAS(G12C)-GDC6036) is presented by, or forms a complex with, a MHC molecule. In some embodiments, the MHC is an HLA and the HLA is HLA- A* 02 : 01 , HL A- A* 03 : 01 , or HL A- A* 11 :01.
The present disclosure provides methods of deep mutational scanning that may be used to determine alternative residues. Deep mutational scanning can be used to identify binding partners with stronger binding affinities. Mutational scanning can comprise modifying (e.g., substituting) 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more than 10 residues of an amino acid sequence described herein. The mutations determined from the mutational scanning may be at any position in an amino acid sequence. In some embodiments, the mutations can be at a CDR region. In some embodiments, the mutation or mutations may be within a CDR1, CDR2, CDR3 or any combination thereof in a VL chain of a binding partner described herein. In some embodiments, the mutation or mutations may be within a CDR1, CDR2, CDR3 or any combination thereof in a VH chain of a binding partner described herein In some embodiments, the binding partner may comprise a mutation (e.g., a single-point mutation) at least one position. The mutation may be in a variable light (VL) chain region and/or in a variable heavy (VH) chain region. In some embodiments, a VL of RM 010 may comprise a sequence as set forth in (positions at which mutations may occur are underlined): DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLOPEDFATYYCOQSSWLYWLVTFGOGTKVEIKRTVGGLE (SEQ ID NO: 257). In some embodiments, a VH of RM 010 may comprise a sequence as set forth in (positions at which mutations may occur are underlined):
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFQWYAMDYWGQGTL VTVSS (SEQ ID NO: 258). In some embodiments, a mutation of the deep mutational scanning can comprise at least one mutation within a LC CDR1 as set forth in any one of SEQ ID NOs: 6, 14, 22, 30, 38, 46, 54, 62, 70, 78, 86, 94, 102, 110, 118, 126, 134, 142, 150, 158, 166, 174, 182, 190, 198, 206, 214, 222, 230, 238, 246, or 254. In some embodiments, a mutation of the deep mutational scanning can comprise at least one mutation within a LC CDR2 as set forth in any one of SEQ ID NOs: 7, 15, 23, 31, 39, 47, 55, 63, 71, 79, 87, 95, 103, 111, 119, 127, 135, 143, 151, 159, 167, 175, 183, 191, 199, 207, 215, 223, 231, 239, 247, or 255. In some embodiments, a mutation of the deep mutational scanning can comprise at least one mutation within a LC CDR3 as set forth in any one of SEQ ID NOs: 8, 16, 24, 32, 40, 48, 56, 64, 72, 80, 88, 96, 104, 112, 120, 128, 136, 144, 152, 160, 168, 176, 184, 192, 200, 208, 216, 224, 232, 240, 248, or 256. In some embodiments, a mutation of the deep mutational scanning can comprise at least one mutation within a HC CDR1 as set forth in any one of SEQ ID NOs: 3, 11, 19, 27, 35, 43, 51, 59, 67, 75, 83, 91, 99, 107, 115, 123, 131, 139, 147, 155, 163, 171, 179, 187, 195, 203, 211, 219, 227, 235, 243, or 251. In some embodiments, a mutation of the deep mutational scanning can comprise at least one mutation within a HC CDR2 as set forth in any one of SEQ ID NOs: 4, 12, 20, 28, 36, 44, 52, 60, 68, 76, 84, 92, 100, 108, 116, 124, 132, 140, 148, 156, 164, 172, 180, 188, 196, 204, 212, 220, 228, 236, 244, or 252. In some embodiments, a mutation of the deep mutational scanning can comprise at least one mutation within a HC CDR3 as set forth in any one of SEQ ID NOs: 5, 13, 21, 29, 37, 45, 53, 61, 69, 77, 85, 93, 101, 109, 117, 125, 133, 141, 149, 157, 165, 173, 181, 189, 197, 205, 213, 221, 229, 237, 245, or 253.
In some embodiments, a mutation of the deep mutational scanning can comprise a mutation in an amino acid sequence as set forth in a HC CDR1 of RM 010 (SEQ ID NO: 75), a HC CDR2 of RM 010 (SEQ ID NO: 76), a HC CDR3 of RM 010 (SEQ ID NO: 77), a LC CDR1 of RM 010 (SEQ ID NO: 78), a LC CDR2 of RM 010 (SEQ ID NO: 79), or a LC CDR3 of RM 010 (SEQ ID NO: 80), or any combination thereof.
In some embodiments, mutations may be determined by mutational scanning of a binding partner described herein. Deep mutational scanning can comprise forming a yeastdisplay library, (e.g., forming a yeast-display library in the scFv format). The yeast-display library can contain variants in which a single position can be diversified. The yeast display library can be subjected to FACS using an antigen of interest. The FACS can enrich a pool of clones that bind the antigen and a pool of clones that may not bind the antigen. The DNA sequences of the enriched pools can then be determined, and an amino acid substitution or multiple amino acid substitutions can be deduced.
A summary of single-point mutations is found in Table E.
Table E. Single-point mutations of RM 010 from deep mutational scanning.
In some embodiments, the binding partner described herein can comprise a polypeptide with an antigen-binding domain or fragment thereof. The antigen-binding domains of the present disclosure can comprise, but are not limited to, the following examples.
In some embodiments, the VH comprises a HC CDR1 having a sequence of ISSSSIH, a HC CDR2 having a sequence of SISSYYGSTSYADSVKG, and a HC CDR3 having a sequence of LWASGLDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQWNWGWPLIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTIS S S SMWVRQAPGKGLEWVASIS S YYGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWGQGTLV TVSS (SEQ ID NO.: 2). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTIS S S SMWVRQAPGKGLEWVASIS S YYGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWGQGTLV TVSS (SEQ ID NO.: 2). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTIS S S SMWVRQAPGKGLEWVASIS S YYGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWGQGTLV TVSS (SEQ ID NO.: 2). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV (SEQ ID NO.: 1). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV (SEQ ID NO.: 1). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV (SEQ ID NO.: 1).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSYYGSTSYADSVKG, and a HC CDR3 having a sequence of LWASGLDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQWNWGWPLIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S YYGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWGQGTLV TVSS (SEQ ID NO: 10). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S YYGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWGQGTLV TVSS (SEQ ID NO: 10). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S YYGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWGQGTLV TVSS (SEQ ID NO: 10). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV (SEQ ID NO: 9). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV (SEQ ID NO: 9). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV (SEQ ID NO: 9).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of SYYGFWQALWALDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQWWYGSPLFT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALWALDYW GQGTLVTVSS (SEQ ID NO: 18). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALWALDYW GQGTLVTVSS (SEQ ID NO: 18). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALWALDYW GQGTLVTVSS (SEQ ID NO: 18). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRT (SEQ ID NO: 17). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRT (SEQ ID NO: 17). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRT (SEQ ID NO: 17).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSYSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of SYYGFWQALWALDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQWWYGSPLFT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALWALDYW GQGTLVTVSS (SEQ ID NO: 26). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S YSIHWVRQAPGKGLEW VASIS S S SGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALWALDYW GQGTLVTVSS (SEQ ID NO: 26). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGL VQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALWALDYW GQGTLVTVSS (SEQ ID NO: 26). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRT (SEQ ID NO: 25). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRT (SEQ ID NO: 25). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRT (SEQ ID NO: 25).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSYYSIH, a HC CDR2 having a sequence of SISSYYGSTYYADSVKG, and a HC CDR3 having a sequence of GYYYPYYAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQGKTYYPIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 34). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 34). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 34). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 33). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 33). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 33).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSYYSIH, a HC CDR2 having a sequence of SISPYYGSTYYADSVKG, and a HC CDR3 having a sequence of GYYYPYYAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQGKTYYPIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISPYYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 42). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISPYYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 42). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISPYYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 42). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 41). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 41). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 41).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSYYSIH, a HC CDR2 having a sequence of SISSYYGSTYYADSVKG, and a HC CDR3 having a sequence of GYYYPYYAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQTSWYHSLIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 50). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 50). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 50). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQTSWYHSLITFGQGTKVEIKRTV (SEQ ID NO: 49). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQTSWYHSLITFGQGTKVEIKRTV (SEQ ID NO: 49). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQTSWYHSLITFGQGTKVEIKRTV (SEQ ID NO: 49).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSYYSIH, a HC CDR2 having a sequence of SISSYYGSTYYADSVKG, and a HC CDR3 having a sequence of GYYYPYYAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQGKTYYPIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVASISSYYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 58). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVASISSYYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 58). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVASISSYYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 58). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 57). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 57). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 57). In some embodiments, the VH comprises a HC CDR1 having a sequence of FSYYSIH, a HC CDR2 having a sequence of SISSYYGSTYYADSVKG, and a HC CDR3 having a sequence of GYYYPYYAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSWWSYPLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 66). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 66). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDYWG QGTLVTVSS (SEQ ID NO: 66). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 65). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 65). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 65).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FQWYAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSSWLYWLVT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSMWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFQWYAMDYWGQGTL VTVSS (SEQ ID NO: 74). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFQWYAMDYWGQGTL VTVSS (SEQ ID NO: 74). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFQWYAMDYWGQGTL VTVSS (SEQ ID NO: 74). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWLVTFGQGTKVEIKRTV (SEQ ID NO: 73). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWLVTFGQGTKVEIKRTV (SEQ ID NO: 73). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWLVTFGQGTKVEIKRTV (SEQ ID NO: 73).
In some embodiments, the VH comprises a HC CDR1 having a sequence of ISSYSIH, a HC CDR2 having a sequence of YISSYSGYTSYADSVKG, and a HC CDR3 having a sequence of GYGWGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQWNWGWPLIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTIS SYSMWVRQAPGKGLEWVAYISS YSGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYGWGMDYWGQGT LVTVSS (SEQ ID NO: 82). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTIS S YSIHWVRQAPGKGLEWVAYIS S YSGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYGWGMDYWGQGT LVTVSS (SEQ ID NO: 82). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTIS S YSIHWVRQAPGKGLEWVAYIS S YSGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYGWGMDYWGQGT LVTVSS (SEQ ID NO: 82). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV (SEQ ID NO: 81). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV (SEQ ID NO: 81). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV (SEQ ID NO: 81).
In some embodiments, the VH comprises a HC CDR1 having a sequence of ISYSSIH, a HC CDR2 having a sequence of YISSSSGYTSYADSVKG, and a HC CDR3 having a sequence of SGYYSSHWYLQSWYQAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSSAWYPVT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence
EVQLVESGGGLVQPGGSLRLSCAASGFTISYSSIHWVRQAPGKGLEWVAYISSSSGYT SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQSWYQ AMDYWGQGTLVTVSS (SEQ ID NO: 90). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTISYSSIHWVRQAPGKGLEWVAYISSSSGYT SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQSWYQ AMDYWGQGTLVTVSS (SEQ ID NO: 90). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGLVQPGGSLRLSCAASGFTISYSSIHWVRQAPGKGLEWVAYISSSSGYT SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQSWYQ AMDYWGQGTLVTVSS (SEQ ID NO: 90). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV (SEQ ID NO: 89). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV (SEQ ID NO: 89). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV (SEQ ID NO: 89).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSYYSH4, a HC CDR2 having a sequence of SISSYYGSTYYADSVKG, and a HC CDR3 having a sequence of SGYYSSHWYLQSWYQAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SAS SLYS, and a LC CDR3 having a sequence of QQASYGPIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF SYYSIHWVRQAPGKGLEWVASISS YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQSWY QAMDYWGQGTLVTVSS (SEQ ID NO: 98). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQSWY QAMDYWGQGTLVTVSS (SEQ ID NO: 98). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQSWY QAMDYWGQGTLVTVSS (SEQ ID NO: 98). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYGPITFGQGTKVEIKRTV (SEQ ID NO: 97). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYGPITFGQGTKVEIKRTV (SEQ ID NO: 97). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYGPITFGQGTKVEIKRTV (SEQ ID NO: 97).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSYSSIH, a HC CDR2 having a sequence of SISSYYGSTYYADSVKG, and a HC CDR3 having a sequence of HYSEKWWGWYTMYIDAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQWWSSSQLIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVASISSYYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARHYSEKWWGWYTMYI DAMDYWGQGTLVTVSS (SEQ ID NO: 106). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVASISSYYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARHYSEKWWGWYTMYI DAMDYWGQGTLVTVSS (SEQ ID NO: 106). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVASISSYYGS TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARHYSEKWWGWYTMYI DAMDYWGQGTLVTVSS (SEQ ID NO: 106). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWSSSQLITFGQGTKVEIKRTV (SEQ ID NO: 105). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWSSSQLITFGQGTKVEIKRTV (SEQ ID NO: 105). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWSSSQLITFGQGTKVEIKRTV (SEQ ID NO: 105).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSYYSIH, a HC CDR2 having a sequence of YISPYSGYTSYADSVKG, and a HC CDR3 having a sequence of GWYYLGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQGKTYYPIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQ GTLVTVSS (SEQ ID NO: 114). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQ GTLVTVSS (SEQ ID NO: 114). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQ GTLVTVSS (SEQ ID NO: 114). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 113). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 113). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 113).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSSYSH4, a HC CDR2 having a sequence of SISPYSSYTSYADSVKG, and a HC CDR3 having a sequence of GWYYLGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQGKTYYPIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQG TLVTVSS (SEQ ID NO: 122). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQG TLVTVSS (SEQ ID NO: 122). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQG TLVTVSS (SEQ ID NO: 122). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 121). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 121). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV (SEQ ID NO: 121).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSYYSIH, a HC CDR2 having a sequence of YISPYSGYTSYADSVKG, and a HC CDR3 having a sequence of GWYYLGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSWWSYPLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQ GTLVTVSS (SEQ ID NO: 130). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQ GTLVTVSS (SEQ ID NO: 130). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQ GTLVTVSS (SEQ ID NO: 130). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 129). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 129). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 129).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSSYSH4, a HC CDR2 having a sequence of YISPYSGYTSYADSVKG, and a HC CDR3 having a sequence of GWYYLGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSWWSYPLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPYSGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQG TLVTVSS (SEQ ID NO: 138). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPYSGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQG TLVTVSS (SEQ ID NO: 138). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPYSGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQG TLVTVSS (SEQ ID NO: 138). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 137). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 137). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 137).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSYSIH, a HC CDR2 having a sequence of SISPYSSYTSYADSVKG, and a HC CDR3 having a sequence of GWYYLGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSWWSYPLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISPYSSYT SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGT LVTVSS (SEQ ID NO: 146). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISPYSSYT SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGT LVTVSS (SEQ ID NO: 146). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISPYSSYT SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQGT LVTVSS (SEQ ID NO: 146). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 145). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 145). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 145).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSYYSIH, a HC CDR2 having a sequence of YISPYSGYTSYADSVKG, and a HC CDR3 having a sequence of GWYYLGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSSAWYPVT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQ GTLVTVSS (SEQ ID NO: 154). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQ GTLVTVSS (SEQ ID NO: 154). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPYSG YTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWGQ GTLVTVSS (SEQ ID NO: 154). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV (SEQ ID NO: 153). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV (SEQ ID NO: 153). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV (SEQ ID NO: 153).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSSYSIH, a HC CDR2 having a sequence of SISPYSSYTSYADSVKG, and a HC CDR3 having a sequence of YWYYMGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSWWSYPLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 162). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 162). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 162). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 161). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 161). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 161).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSSYSIH, a HC CDR2 having a sequence of SISPYSSYTSYADSVKG, and a HC CDR3 having a sequence of YWYYMGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSSAWYPVT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 170). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 170). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 170). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV (SEQ ID NO: 169). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV (SEQ ID NO: 169). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV (SEQ ID NO: 169).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSSYSIH, a HC CDR2 having a sequence of SISPYSSYTSYADSVKG, and a HC CDR3 having a sequence of YWYYMGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSSYYEELIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 178). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 178). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 178). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSYYEELITFGQGTKVEIKRTV (SEQ ID NO: 177). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSYYEELITFGQGTKVEIKRTV (SEQ ID NO: 177). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSYYEELITFGQGTKVEIKRTV (SEQ ID NO: 177).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSSYSIH, a HC CDR2 having a sequence of SISPYSSYTSYADSVKG, and a HC CDR3 having a sequence of YWYYMGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQAYSDPLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 186). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 186). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 186). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQAYSDPLTFGQGTKVEIKRTV (SEQ ID NO: 185). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQAYSDPLTFGQGTKVEIKRTV (SEQ ID NO: 185). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQAYSDPLTFGQGTKVEIKRTV (SEQ ID NO: 185).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSSYSIH, a HC CDR2 having a sequence of SISPYSSYTSYADSVKG, and a HC CDR3 having a sequence of YWYYMGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQGSSSLLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 194). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 194). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 194). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV (SEQ ID NO: 193). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV (SEQ ID NO: 193). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV (SEQ ID NO: 193).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSSYSIH, a HC CDR2 having a sequence of SISPYSSYTSYADSVKG, and a HC CDR3 having a sequence of YWYYMGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSGSYLLIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 202). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 202). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 202). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSGSYLLITFGQGTKVEIKRTV (SEQ ID NO: 201). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSGSYLLITFGQGTKVEIKRTV (SEQ ID NO: 201). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSGSYLLITFGQGTKVEIKRTV (SEQ ID NO: 201).
In some embodiments, the VH comprises a HC CDR1 having a sequence of VSSYSIH, a HC CDR2 having a sequence of SISPYSSYTSYADSVKG, and a HC CDR3 having a sequence of YWYYMGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQADYEFGLIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 210). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 210). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYSSY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWGQG TLVTVSS (SEQ ID NO: 210). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQADYEFGLITFGQGTKVEIKRTV (SEQ ID NO: 209). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQADYEFGLITFGQGTKVEIKRTV (SEQ ID NO: 209). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQADYEFGLITFGQGTKVEIKRTV (SEQ ID NO: 209). In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSYSIH, a HC CDR2 having a sequence of YISSSYGYTSYADSVKG, and a HC CDR3 having a sequence of SSRQYYHSQVEPPMAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSWWSYPLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 218). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 218). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 218). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 217). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 217). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 217). In some embodiments, the VH comprises a HC CDR1 having a sequence of FSYYSIH, a HC CDR2 having a sequence of YISSSYGYTSYADSVKG, and a HC CDR3 having a sequence of SSRQYYHSQVEPPMAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSWWSYPLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 226). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 226). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 226). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 225). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 225). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 225). In some embodiments, the VH comprises a HC CDR1 having a sequence of VSSSSIH, a HC CDR2 having a sequence of YISSSYGYTSYADSVKG, and a HC CDR3 having a sequence of SSRQYYHSQVEPPMAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSWWSYPLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 234). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 234). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 234). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 233). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 233). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 233). In some embodiments, the VH comprises a HC CDR1 having a sequence of VSYSSIH, a HC CDR2 having a sequence of YISSSYGYTSYADSVKG, and a HC CDR3 having a sequence of SSRQYYHSQVEPPMAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSWWSYPLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 242). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 242). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVAYISSSYGY TSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPPMA MDYWGQGTLVTVSS (SEQ ID NO: 242). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 241). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 241). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV (SEQ ID NO: 241). In some embodiments, the VH comprises a HC CDR1 having a sequence of VSSYSIH, a HC CDR2 having a sequence of YISPYSGYTYYADSVKG, and a HC CDR3 having a sequence of YWYYLGMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQGSSSLLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPYSGY TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYLGMDYWGQG TLVTVSS (SEQ ID NO: 250). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPYSGY TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYLGMDYWGQG TLVTVSS (SEQ ID NO: 250). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPYSGY TYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYLGMDYWGQG TLVTVSS (SEQ ID NO: 250). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV (SEQ ID NO: 249). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV (SEQ ID NO: 249). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV (SEQ ID NO: 249).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FYDLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQASWLYWLIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSMWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMDYWGQGTLV TVSS (SEQ ID NO.: 270). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMDYWGQGTLV TVSS (SEQ ID NO.: 270). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMDYWGQGTLV TVSS (SEQ ID NO.: 270). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 269). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 269). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 269).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FLDLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQASWLYWKIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFLDLAMDYWGQGTLV TVSS (SEQ ID NO.: 278). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFLDLAMDYWGQGTLV TVSS (SEQ ID NO.: 278). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFLDLAMDYWGQGTLV TVSS (SEQ ID NO.: 278). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKITFGQGTKVEIK (SEQ ID
NO.: 277). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKITFGQGTKVEIK (SEQ ID NO.: 277). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKITFGQGTKVEIK (SEQ ID NO.: 277).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FTVLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQASWLYWLIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 286). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 286). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 286). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 285). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 285). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 285).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FTVLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQASWLYWKVT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 294). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 294). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 294). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKVTFGQGTKVEIK (SEQ ID NO.: 293). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKVTFGQGTKVEIK (SEQ ID NO.: 293). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKVTFGQGTKVEIK (SEQ ID NO.: 293).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FTVLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQASWLYWVIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 302). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSMWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 302). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 302). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWVITFGQGTKVEIK (SEQ ID NO.: 301). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWVITFGQGTKVEIK (SEQ ID NO.: 301). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWVITFGQGTKVEIK (SEQ ID NO.: 301).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FHDLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQASWLYWKIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHDLAMDYWGQGTLV TVSS (SEQ ID NO.: 310). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHDLAMDYWGQGTLV TVSS (SEQ ID NO.: 310). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHDLAMDYWGQGTLV TVSS (SEQ ID NO.: 310). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKITFGQGTKVEIK (SEQ ID NO.: 309). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKITFGQGTKVEIK (SEQ ID NO.: 309). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKITFGQGTKVEIK (SEQ ID NO.: 309).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FHELAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQASWLYWLIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMDYWGQGTLV TVSS (SEQ ID NO.: 318). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMDYWGQGTLV TVSS (SEQ ID NO.: 318). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSMWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMDYWGQGTLV TVSS (SEQ ID NO.: 318). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 317). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 317). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 317).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FTVLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSSWLYWKIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 326). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 326). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 326). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKVEIK (SEQ ID NO.: 325). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKVEIK (SEQ ID NO.: 325). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKVEIK (SEQ ID NO.: 325).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FELLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSSWLYWKIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFELLAMDYWGQGTLV TVSS (SEQ ID NO.: 334). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFELLAMDYWGQGTLV TVSS (SEQ ID NO.: 334). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFELLAMDYWGQGTLV TVSS (SEQ ID NO.: 334). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKVEIK (SEQ ID NO.: 333). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKVEIK (SEQ ID NO.: 333). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKVEIK (SEQ ID NO.: 333).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FTVLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSSWLYWKVT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 342). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 342). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 342). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKVTFGQGTKVEIK (SEQ ID NO.: 341). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKVTFGQGTKVEIK (SEQ ID NO.: 341). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKVTFGQGTKVEIK (SEQ ID NO.: 341).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FYDLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSSWLYWKVT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMDYWGQGTLV TVSS (SEQ ID NO.: 350). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMDYWGQGTLV TVSS (SEQ ID NO.: 350). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMDYWGQGTLV TVSS (SEQ ID NO.: 350). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKVTFGQGTKVEIK (SEQ ID NO.: 349). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKVTFGQGTKVEIK (SEQ ID NO.: 349). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKVTFGQGTKVEIK (SEQ ID NO.: 349).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FHELAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQASWLYWLIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMDYWGQGTLV TVSS (SEQ ID NO.: 358). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMDYWGQGTLV TVSS (SEQ ID NO.: 358). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMDYWGQGTLV TVSS (SEQ ID NO.: 358). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 357). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 357). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 357).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FTVLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQASWLYWKVT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 366). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 366). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMDYWGQGTLV TVSS (SEQ ID NO.: 366). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKVTFGQGTKVEIK (SEQ ID NO.: 365). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKVTFGQGTKVEIK (SEQ ID NO.: 365). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKVTFGQGTKVEIK (SEQ ID NO.: 365).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FYDLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSSWLYWVIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMDYWGQGTLV TVSS (SEQ ID NO.: 374). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMDYWGQGTLV TVSS (SEQ ID NO.: 374). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMDYWGQGTLV TVSS (SEQ ID NO.: 374). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWVITFGQGTKVEIK (SEQ ID NO.: 373). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWVITFGQGTKVEIK (SEQ ID NO.: 373). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWVITFGQGTKVEIK (SEQ ID NO.: 373).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FHELAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQASWLYWLIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMDYWGQGTLV TVSS (SEQ ID NO.: 382). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMDYWGQGTLV TVSS (SEQ ID NO.: 382). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMDYWGQGTLV TVSS (SEQ ID NO.: 382). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 381). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 381). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKVEIK (SEQ ID NO.: 381).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FAHLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSSWLYWLIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFAHLAMDYWGQGTLV TVSS (SEQ ID NO.: 390). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFAHLAMDYWGQGTLV TVSS (SEQ ID NO.: 390). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFAHLAMDYWGQGTLV TVSS (SEQ ID NO.: 390). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWLITFGQGTKVEIK (SEQ ID NO.: 389). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWLITFGQGTKVEIK (SEQ ID NO.: 389). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWLITFGQGTKVEIK (SEQ ID NO.: 389). In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FEDLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQASWLYWYIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFEDLAMDYWGQGTLV TVSS (SEQ ID NO.: 398). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFEDLAMDYWGQGTLV TVSS (SEQ ID NO.: 398). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFEDLAMDYWGQGTLV TVSS (SEQ ID NO.: 398). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWYITFGQGTKVEIK (SEQ ID NO.: 397). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWYITFGQGTKVEIK (SEQ ID NO.: 397). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWYITFGQGTKVEIK (SEQ ID NO.: 397).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FRTLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSSWLYWIIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSMWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFRTLAMDYWGQGTLV TVSS (SEQ ID NO.: 406). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFRTLAMDYWGQGTLV TVSS (SEQ ID NO.: 406). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SMWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFRTLAMDYWGQGTLV TVSS (SEQ ID NO.: 406). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWIITFGQGTKVEIK (SEQ ID NO.: 405). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWIITFGQGTKVEIK (SEQ ID NO.: 405). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWIITFGQGTKVEIK (SEQ ID NO.: 405).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FTHYAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQSSWLYWKIT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTHYAMDYWGQGTLV TVSS (SEQ ID NO.: 414). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTHYAMDYWGQGTLV TVSS (SEQ ID NO.: 414). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTHYAMDYWGQGTLV TVSS (SEQ ID NO.: 414). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKVEIK (SEQ ID NO.: 413). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKVEIK (SEQ ID NO.: 413). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKVEIK (SEQ ID NO.: 413).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FRLYAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQASWLHWKLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFRLYAMDYWGQGTLV TVSS (SEQ ID NO.: 422). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFRLYAMDYWGQGTLV TVSS (SEQ ID NO.: 422). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFRLYAMDYWGQGTLV TVSS (SEQ ID NO.: 422). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLHWKLTFGQGTKVEIK (SEQ ID NO.: 421). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLHWKLTFGQGTKVEIK (SEQ ID NO.: 421). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLHWKLTFGQGTKVEIK (SEQ ID NO.: 421).
In some embodiments, the VH comprises a HC CDR1 having a sequence of FSSSSIH, a HC CDR2 having a sequence of SISSSSGSTSYADSVKG, and a HC CDR3 having a sequence of FHRLAMDY. In some embodiments, the VL comprises a LC CDR1 having a sequence of RASQSVSSAVA, a LC CDR2 having a sequence of SASSLYS, and a LC CDR3 having a sequence of QQASWLHWKLT. In some embodiments, the VH comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence EVQLVESGGGL VQPGGSLRLSCAASGFTFSSSSIHWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHRLAMDYWGQGTLV TVSS (SEQ ID NO.: 430). In some embodiments, the VH comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHRLAMDYWGQGTLV TVSS (SEQ ID NO.: 430). In some embodiments, the VH comprises an amino acid sequence as set forth in the amino acid sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHRLAMDYWGQGTLV TVSS (SEQ ID NO.: 430). In some embodiments, the VL comprises an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLHWKLTFGQGTKVEIK (SEQ ID NO.: 429). In some embodiments, the VL comprises an amino acid sequence having at least about 80% sequence identity to the amino acid sequence
DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLHWKLTFGQGTKVEIK (SEQ ID NO.: 429). In some embodiments, the VL comprises an amino acid sequence as set forth in the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLHWKLTFGQGTKVEIK (SEQ ID NO.: 429).
In embodiments, the binding partner binds to a protein in its native form, with the exception that the drug or other molecule is covalently attached to it. “Native form” means the intact protein that retains its biological function before covalent attachment of the drug or other molecule. In embodiments, the native form or the protein is its form before being fragmented such as by intracellular processing. In embodiments, the binding proteins therefore bind to full length polypeptides that are covalently attached to the drug or other molecule and wherein the covalently bound drug or other molecule at least in part permits the preferential binding of the binding partners. In general, a polypeptide, which is used interchangeably herein with the term “protein,” comprises more than 50 contiguous amino acids. In embodiments, a binding partner binds with specificity to an intact protein that is covalently attached to a drug or other molecule. In other embodiments, the binding partners bind with specificity to a peptide comprising the covalently bound molecule. In embodiments, the binding partner binds with specificity to a peptide having a specific amino acid sequence and is covalently conjugated to another molecule, such as a drug. In embodiments, the binding partner binds preferentially to a peptide covalently bound to a molecule such as a drug, where the sequence of the peptide is not relevant. This preferential binding is relative to binding to the same peptide that is not conjugated to the drug. In embodiments, the binding partner binds preferentially to a peptide comprising a KRAS(G12) mutation, or to a variant thereof, wherein the variant is at least 50% similar to the KRAS(G12)-containing peptide. This preferential binding is relative to binding to a KRAS(G12)-containing peptide, or the variant thereof, respectively, that is not covalently conjugated to the drug or other molecule.
In embodiments, the described binding partners bind with specificity to peptide conjugates that are of suitable length to be presented in a major histocompatibility complex (MHC), referred to as human leukocyte antigen (HLA) in humans, or to MHC or its equivalent complex in non-human animals, including but not limited to non-human mammals.
In general, the peptide conjugate comprises fewer than 50 contiguous amino acids. In embodiments, the peptide conjugates which comprise the described epitopes may therefore be from 2-49 amino acids in length. In embodiments, the peptide to which the drug or other molecule is covalently attached, and which attached drug and one or more residues of the peptide may be comprised by the epitope, comprises from 4-12 contiguous amino acids, which may or may not be derived from a longer protein during the processing of a protein, such as an antigen processed for presentation by an MHC molecule. In embodiments, the drug is conjugated to a peptide that comprises, or consists of 7-30 amino acids. In embodiments, the drug or other molecule is conjugated to a peptide that comprises, or consists of, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids, and which may be presented in an MHC Class I context. In embodiments, the drug or other molecule is conjugated to a peptide that is 9-30 amino acids, inclusive, and including all numbers and ranges of numbers there between, and which may be presented in an MHC Class II context. In embodiments, the drug or other molecule is conjugated to a peptide comprises at least 7 amino acids. Non-classical MHC class I molecules function to mediate inhibitory or activating stimuli in natural killer (NK) cells. Non-classical MHC class I molecules can be expressed by immune and tumor cells. For example, expression of non-classical MHC complexes on malignant cells hampers cytotoxic activity of effector cells in the immune system. Overexpression of non-classical MHC class I molecules, including but not limited to HLA-E, HLA-F, and HLA-G, can be found in cancer cells. In some embodiments, the drug or other molecule is conjugated to a peptide that is 9-30 amino acids, inclusive, and including all numbers and ranges of numbers there between, and which may be presented in a non-classical MHC Class I context. In some embodiments, the peptide conjugate forms a complex with a non-classical MHC class I molecule. In some embodiments, the non-classical MHC class I molecule is selected from the group consisting of HLA-E, HLA-F, and HLA-G. In some embodiments, the non-classical MHC class I molecule is selected from HLA-E, HLA-F, HLA-G, or some combination thereof.
In embodiments, a binding partner binds with specificity to a peptide conjugate that is covalently conjugated to a drug or other molecule independent of MHC presentation. In embodiments, the binding partner binds with specificity to the peptide conjugate only when the peptide conjugate is presented by an MHC molecule. In embodiments, the binding partner can bind with specificity to a peptide conjugate in both an MHC -independent and an MHC- presented context. In embodiments, the MHC-peptide conjugate complex comprises an antigen to which a described binding partner binds with specificity.
In embodiments, a described binding partner exhibits at least one improved property relative to the same property of a reference binding partner. In embodiments, a reference binding partner is RM 010. In one embodiment, a described binding partner exhibits a greater affinity for its target relative to a reference binding partner.
In embodiments, the binding partners accordingly can bind to cells via any MHC that can present peptide conjugates. In embodiments, the HLA is expressed by cells that are capable of Class I, Class II, or Class III MHC presentation. In embodiments, the binding partners can bind to cells that express Class I MHC that presents the peptide conjugate. Those skilled in the art will recognize that Class I MHC includes, among other components, a polymorphic a chain and 02 microglobulin, wherein the peptide conjugate binds to the polymorphic chain.
In embodiments, the cells are antigen presenting cells (APCs). In embodiments, the cells are so-called professional antigen presenting cells, and thus may include but are not limited to macrophages and dendritic cells, which display Class II MHC. Those skilled in the art will recognize that Class II MHC includes, among other components, MHC polymorphic a and 0 chains, and the displayed peptide conjugate binds to both chains. In other embodiments, Class II MHC may be displayed with the peptide conjugate by other cell types, such as cancer/tumor cells, and thus the disclosure provides for direct recognition of such cells using the described binding partners, without requirement for a professional APC.
In embodiments, the peptide conjugate is displayed by a non-classical MHC complex, which may include CD Id, MR1, MHC-E, -F, -G and/or other emerging family members that will be recognized by those skilled in the art.
The disclosure includes binding partners that bind with specificity to a peptide conjugate displayed only by a specific MHC type, and thus provides binding partners that discriminate between MHC types.
In embodiments, a binding partner of this disclosure can bind with specificity to a peptide conjugate comprising a covalently conjugated drug or other molecule that is displayed by more than one specific MHC type. In embodiments, a single binding partner of the disclosure is suitable for use with a diversity of HLA types. Furthermore, a single binding partner can bind with specificity multiple combinations of (a) peptide conjugate(s) comprising a covalently conjugated drug or other molecule and (b) HLA types. In some embodiments, the antigen-binding domain described herein can bind to different peptides conjugated to AMG-510, MRTX849, or GDC6036 that are presented by different HLAs having distinct preferences for peptide binding. For example, compare the HLA-A*03/l 1 peptide, VVVGAC*GVGK with KLVVVGAC*GV, which shows that the antibody RA DI 1 still selectively binds to the hapten-peptide-HLA complex relative to binding to the free drug. In embodiments, a binding partner of this disclosure can bind with specificity to a peptide conjugate comprising a covalently conjugated drug only in a specific MHC context. In embodiments, the peptide conjugate is displayed by an MHC class I type selected from HLA- A, -B, -C, and combinations thereof. In certain aspects, the peptide conjugate is displayed in the context of any MHC class I that is A*02. Antibodies provided herein can bind to KRAS(G12C)-AMG-510 conjugate, KRAS(G12C)-MRTX849 conjugate, and/or KRAS(G12C)-GDC6036 conjugate presented on HLA-A*02:01, HLA-A*03:01 and HLA- A* 11 :01. In non-limiting embodiments, the disclosure provides scDbs that are specific for a particular drug that is covalently bound to a described peptide that is present on a specific HLA, or the same drug that is covalently bound to a described peptide that is present on two different HLAs.
In embodiments, the disclosure comprises selecting an individual based on the HLA type of the individual and selecting an antibody described herein for treating that individual. In embodiments, the binding partner is selected based at least on part on the degree of the HLA restriction exhibited by the selected binding partner, relative to the HLA type of the individual.
In certain embodiments, such as KRAS(G12C) binding partners, representative examples are provided below and bind to two different peptides derived from KRAS(G12C) conjugated to a drug (AMG-510, also referred to as sotorasib; MRTX849, also referred to as adagrasib; GDC6036, also referred to as divarasib, or any combination thereof).
In embodiments, the peptide conjugate is displayed by cells that participate in, or can be the targets of, cell-mediated immune responses. In embodiments the peptide conjugate that is displayed in any suitable MHC context is comprised by a cell that is recognized by a leukocyte, including but not necessarily limited to a T cell or a natural killer (NK) cell. In embodiments, the T cell is a CD4+ T cell, a CD8+ T cell, a double positive CD4+/CD8+ T cell, a CD4+/CD8+ double negative T cell, a NKT cell, or a y5 T cell. Thus, and as described further below, the disclosure provides binding partners that are configured to interact with both the presented peptide conjugate and cells that participate in cell-mediated immune responses. In embodiments, certain described binding partners are capable of binding to a complex of 1) a specific MHC and 2) a specific peptide conjugate. In embodiments, certain described binding partners are capable of being bound to a specific peptide conjugate presented by at least two different MHCs.
In embodiments, any binding partner of this disclosure comprises at least one chain that comprises a complementary determining region (CDR) that is CDR1, CDR2, or CDR3 from any heavy or light chain amino acid sequence described herein. In certain examples in the present specification, the CDRs are shown in bold font. The amino acid sequences of the CDR sequences are separately encompassed by this disclosure by way of their positions in the described heavy and light chain amino acid sequences. The disclosure includes binding partners that comprise a described heavy chain CDR1, CDR2, and CDR3. The disclosure also includes binding partners that comprise a described light chain CDR1, CDR2, and CDR3. The disclosure also includes binding partners that comprise a described heavy chain CDR1, CDR2, and CDR3 and a described light chain CDR1, CDR2, and CDR3. For amino acid sequences of this disclosure that include amino acids that comprise purification or protein production tags, such as HIS tags and/or AVI-tags, the disclosure includes the proviso that the sequences of the described tags may be excluded from the amino acid sequences. Amino acids between the described tags may also be excluded. Binding partners of this disclosure can be provided as intact immunoglobulins or as fragments of immunoglobulins, including but not necessarily limited to antigen-binding (Fab) fragments, Fab' fragments, (Fab')2 fragments, Fd (N-terminal part of the heavy chain) fragments, Fv fragments (two variable domains), diabodies (Dbs), dAb fragments, single domain fragments or single monomeric variable antibody domains, single-chain Diabodies (scDbs), isolated complementary determining regions (CDRs), single-chain variable fragment (scFv), and other antibody fragments that retain antigen-binding function. In embodiments, one or more binding partners are provided as a component of a Bi-specific T-cell engager (BiTE), bispecific killer cell engager (BiKE), CrossMab (e.g., a binding partner containing four different chains; immunoglobulin crossover (also known as Fab domain exchange or CrossMab format) technology (see e.g., W02009/080253; Schaefer et al., Proc. Natl. Acad. Sci. USA, 108: 11187-11192 (2011).), or a chimeric antigen receptor (CAR), such as for producing chimeric antigen receptor T cells (e.g., CAR T cells) and CAR natural killer (NK) cells, and killer macrophages. The disclosure includes binding partners that include the described heavy and light chain variable regions.
The binding partners of this disclosure can be a scFv. In the present disclosure, the VH of the polypeptide can be linked to the VL through a linker. The linker can be a peptide linker that consists of amino acids such as glycine and/or serine residues used alone or in combination, to link variable heavy and variable light chain regions together. In one embodiment, the flexible polypeptide linker is a Gly/Ser linker and comprises the amino acid sequence (Gly-Gly-Gly-Ser)n, where n is a positive integer equal to or greater than 1. For example, n=l, n=2, n=3, n=4, n=5, n=6, n=7, n=8, n=9 and n=10. In one embodiment, the flexible polypeptide linkers include, but are not limited to, (Gly4Ser)4 or (Gly4Ser)3. In another embodiment, the linkers include multiple repeats of (Gly2Ser), (GlySer), or (GlysSer). In some cases, the linker sequence comprises (G4S)n, wherein n=2 to 4. In some cases, the linker sequence comprises (G4S)n, wherein n=l to 3. In some cases, the linker can comprise (G4S)n or (S4G)n where n is any integer from 1 to 10. In some cases, the linker can comprise the glycine-serine-alanine linker G4SA3 or the glycine-serine linker (G4S
The binding partners of this disclosure can be a Bi-specific T-cell engager (BiTE). BiTE therapies can be used to connect a subject’s endogenous T cells to cancerous cells. A BiTE molecule can comprise two Fv fragments from monoclonal antibodies, joined by a peptide linker. A BiTE molecule can comprise a first antigen-binding domain and a second antigen-binding domain. The first antigen-binding domain can be specific for and bind to a T cell antigen. The second antigen-binding domain can bind to a tumor antigen (e.g., a tumor- associated antigen) expressed on the surface of cancerous cells. In some embodiments, the BiTE molecule can specifically bind to a peptide conjugate/MHC complex and a T cell surface antigen. In some embodiments, the T cell surface antigen is CD3 (e.g., CD3 epsilon, CD3 delta, or CD3 gamma), a TCR alpha chain, a TCR beta chain, a TCR gamma chain, a TCR delta chain, ICOS, CD28, CD27, HVEM, LIGHT, CD40, 4-1BB (CD137), 0X40, DR3, GITR, CD30, TIM1, SLAM (e.g, a SLAM family member), CD2, CD4, CD8, or CD226. In some embodiments, the tumor-associated antigen comprises a peptide conjugate formed by the covalent reaction of a targeted covalent inhibitor with a peptide in a tumor cell. In some embodiments, the tumor-associated antigen comprises a peptide that is a segment of a protein that is associated with cancer, optionally wherein the protein is encoded by a gene that is mutated in a cancer. In some embodiments, the protein is overexpressed in a cancer. In some embodiments, the segment of a protein is overexpressed in cancer. Cancer testis antigens are a class of tumor-associated antigens expressed in human tumors. In some embodiments, the protein is a member of the cancer testis antigen (CTA) family. In some embodiments, the protein associated with cancer comprises an endogenous retrovirus (ERV). In some embodiments, the protein is derived from a retrotransposon family, such as long interspersed elements (LINEs) or short interspersed elements (SINEs). In some embodiments, the tumor- associated antigen is an antigen associated with renal cell carcinoma. In some embodiments, the tumor-associated antigen is an antigen associated with breast cancer. In some embodiments, the tumor-associated antigen is an antigen associated with prostate cancer. In some embodiments, the tumor-associated antigen is an antigen associated with pancreatic cancer. In some embodiments, the tumor-associated antigen is an antigen associated with lung cancer. In some embodiments, the tumor-associated antigen is an antigen associated with liver cancer. In some embodiments, the tumor-associated antigen is an antigen associated with ovarian cancer. In some embodiments, the tumor-associated antigen is an antigen associated with cervical cancer. In some embodiments, the tumor-associated antigen is an antigen associated with colon cancer. In some embodiments, the tumor-associated antigen is an antigen associated with esophageal cancer. In some embodiments, the tumor- associated antigen is an antigen associated with glioma. In some embodiments, the tumor- associated antigen is an antigen associated with glioblastoma or another brain cancer. In some embodiments, the tumor-associated antigen is an antigen associated with stomach cancer. In some embodiments, the tumor-associated antigen is an antigen associated with bladder cancer. In some embodiments, the tumor-associated antigen is an antigen associated with testicular cancer. In some embodiments, the tumor-associated antigen is an antigen associated with head and neck cancer. In some embodiments, the tumor-associated antigen is an antigen associated with melanoma or another skin cancer. In some embodiments, the tumor- associated antigen is an antigen associated with any sarcoma including but not limited to fibrosarcoma, angiosarcoma, osteosarcoma, and rhabdomyosarcoma. In some embodiments, the tumor-associated antigen is an antigen associated with any blood cancer, including all types of leukemia, lymphoma, and myeloma. In some embodiments, the tumor-associated antigen is an antigen associated with endometrial cancer.
The binding partners can comprise a first antigen-binding domain that specifically binds to the peptide-conjugate/MHC complex and a second antigen-binding domain that specifically binds to a T cell surface marker. The binding partners can comprise two polypeptide chains. For example, the polypeptide can comprise a first polypeptide chain comprising the first antigen-binding domain and a second polypeptide chain comprising the second antigen-binding domain. In some cases, the two polypeptide chains can comprise a Fc region. The binding partners of this disclosure can be bispecific or multivalent antibodies or antibody fragments comprising a first antigen-binding domain that specifically binds to the peptide-conjugate/MHC complex and a second antigen-binding domain that specifically binds to a T cell surface marker. In some embodiments, the bispecific or multivalent molecule can comprise a Fc region chosen from heavy chain constant regions of human IgM, IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2. The Fc region can be a modified version of the wildtype Fc region. For example, the Fc region can be a silenced version of the Fc region, including but not limited to, LAL A Fc, LALAPG Fc, LAL AKA Fc, LAGA Fc, LGGR Fc, or LALE Fc. The Fc region can comprise one or more mutations of a wild type Fc region. The Fc region can be linked to one or more tumor targeting moi eties, one or more T cell engagers, or a cytokine molecule. In some embodiments, the interface of a first and second Fc region can be altered to increase or decrease dimerization. The dimerization of the Fc region can be enhanced by providing an Fc interface of a first and a second Fc region with a paired-cavity protuberance, e.g. knob-in-hole. Knob-in-hole as described in US 5,731,116, US 7,476,724 and Ridgway, J. et al. (1996) Prot. Engineering 9(7): 617-621, broadly involves: (1) mutating the CH3 domain of one or both antibodies to promote heterodimerization; and (2) combining the mutated antibodies under conditions that promote heterodimerization. "Knobs" or "protuberances" can be created by replacing a small amino acid in a parental antibody with a larger amino acid (e.g. T366Y or T366W). "Holes" or "cavities" are created by replacing a larger residue in a parental antibody with a smaller amino acid (e.g. Y407T, T366S, L368A, and/or Y407V). Exemplary knob-in-hole mutations include S354C or T366W in the "knob" heavy chain and Y349C, T366S, L368A, or Y407V in the "hole" heavy chain. For bispecific antibodies including an Fc domain, introduction of specific mutations into the constant region of the heavy chains to promote the correct heterodimerization of the Fc portion can be used. These techniques include the knob-in-hole approach which involves the introduction of a bulky residue into one of the CH3 domains of one of the antibody heavy chains. This bulky residue fits into a complementary "hole" in the other CH3 domain of the paired heavy chain so as to promote correct pairing of heavy chains (see e.g., US7642228). In some cases, the two polypeptide chains can be combined using the knob-in-hole approach described herein. In some cases, the two polypeptide chains cannot be combined using the knob-in-hole approach. In some cases, the first antigen-binding domain and the second antigen-binding domain are linked by a linker. The linker can be a peptide linker that consists of amino acids such as glycine and/or serine residues used alone or in combination. The linker can comprise at least about 1, 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, or more amino acid residues in length. In some cases, the linker can comprise (648)11 where n is any integer from 1 to 10. In one embodiment, the flexible polypeptide linker is a Gly/Ser linker and comprises the amino acid sequence (Gly-Gly-Gly-Ser)n, where n is a positive integer equal to or greater than 1. For example, n=l, n=2, n=3, n=4, n=5, n=6, n=7, n=8, n=9 and n=10. In one embodiment, the flexible polypeptide linkers include, but are not limited to, (Gly4Ser)4 or (Gly4Ser)3. In another embodiment, the linkers include multiple repeats of (Gly2Ser), (GlySer) or (GlysSer). In some cases, the linker sequence comprises (G4S)n, wherein n=2 to 4. In some cases, the linker sequence comprises (G4S)n, wherein n=l to 3. In some cases, the linker can comprise (G4S)n or (S4G)n where n is any integer from 1 to 10. In some cases, the linker can comprise the glycine-serine-alanine linker G4SA3 or a glycine-serine linker (648)4.
The first or the second polypeptide chain can further be fused to a cytokine or fragment thereof. The cytokine can comprise IL-2, IL-7, IL-15, IL-12, IL-18, or IL-21, or an interferon (IFN). In some cases, the cytokine can be selected from the group consisting of IFNy, IL-2, IL-4, IL-6, IL-7, IL-10, IL-12, IL-13, IL-15, IL-18, IL-23, IL-24, IL-27, IL-28a, IL-28b, IL-29, KGF, IFN a (e.g., INF a 2b), IFN 0, IFN , and GM-CSF. The cytokines can be modified cytokines or cytokines comprising one or more mutations compared to wild-type cytokines. In some embodiments, the first or the second polypeptide chain can comprise an agonist (e.g., an agonist to CD28 or 4-1BB). In some embodiments, the cytokine is a single chain cytokine. In some embodiments, the cytokine is a multichain cytokine. In some cases, the two polypeptides can contain two moieties. In some cases, the two polypeptide chains can contain at least three moieties. In some cases, the first moiety can bind to the peptide conjugate/MHC complex, the second moiety can comprise a binding partner for a T cell surface antigen (e.g., T cell engager), and the third moiety can comprise a cytokine or fragment thereof. In some cases, the first polypeptide chain can contain the antigen-binding domain for the peptide conjugate/MHC complex and the second polypeptide chain can contain the antigen-binding domain for a T cell surface antigen. In some cases, the first polypeptide chain can contain the antigen-binding domain for the T cell surface antigen and the second polypeptide chain can contain the antigen-binding domain for the peptide conjugate/MHC complex. In some cases, the first polypeptide chain can contain the antigenbinding domain for the peptide conjugate/MHC complex and a cytokine or fragment thereof and the second polypeptide chain can contain the antigen-binding domain for the T cell surface antigen. In some cases, the first polypeptide chain can contain the antigen-binding domain for the T cell surface antigen and a cytokine or fragment thereof and the second polypeptide chain can contain the antigen-binding domain for the peptide conjugate/MHC complex. In some cases, the first polypeptide chain can contain the antigen-binding domain for the peptide conjugate/MHC complex and the second polypeptide chain can contain the antigen-binding domain for the T cell surface antigen and a cytokine or fragment thereof. In some cases, the first polypeptide chain can contain the antigen-binding domain for the T cell surface marker and the second polypeptide chain can contain the antigen-binding domain for the peptide conjugate/MHC complex and a cytokine or fragment thereof. In various cases, the antigen-binding domain for the peptide conjugate/MHC complex and the antigen-binding domain for the T cell surface antigen can be on a same polypeptide chain, and the cytokine or fragment thereof is fused to a different polypeptide chain. In some cases, the antigen-binding domain for the peptide conjugate/MHC complex and the cytokine or fragment thereof can be on a same polypeptide chain, and the antigen-binding domain for the T cell surface antigen is on a different polypeptide chain. In some cases, the antigen-binding domain for the T cell surface antigen and the cytokine or fragment thereof can be on a same polypeptide chain, and the antigen-binding domain for the peptide conjugate/MHC complex is on a different polypeptide chain.
The polypeptide can be an intact antibody, a bispecific antibody, a multispecific antibody, an antigen-binding (Fab) fragment, an Fab’ fragment, an (Fab’)2 fragment, an Fd, an Fv, a dAb, a single domain fragment or single monomeric variable antibody domain, a single-chain Diabody (scDb), a diabody (Db), a dual-affinity retargeting (DART) molecule, a single-chain variable fragment (scFv), a Bi-specific T-cell engager (BiTE), bispecific killer cell engager (BiKE), CrossMab, a tri-specific binding partner, a chimeric antigen receptor (CAR), a monobody (e.g., adnectin), a DARPin, an anticalin, an affibody, a nanobody, or an affimer. In some embodiments, the nanobody is derived from the heavy chain variable domain of antibodies found in members of Camelidae (e.g. camels, llamas, alpacas). In some cases, the polypeptide can be a bispecific antibody. The bispecific antibody can be a bispecific T-cell engager (BiTE). The polypeptide can comprise a first antigen-binding domain and a second antigen-binding domain. The second antigen-binding domain can bind to a T cell surface marker. The T cell surface marker can be CD3 epsilon, CD3 gamma, CD3 delta, a TCR alpha, a TCR beta, a TCR gamma, a TCR delta, ICOS, CD28, CD27, HVEM, LIGHT, CD40, 4-1BB (CD137), 0X40, DR3, GITR, CD30, TIM1, SLAM (e.g, a SLAM family member), CD2, CD4, CD8, or CD226. In some cases, the antigen-binding domain and the second antigen-binding domain of the polypeptide can be linked by a linker. As described above, the linker can comprise at least about 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, or more amino acid residues in length. The linker can comprise (G4S)n or (S4G)n, where n is any integer from 1 to 10.
In some embodiments, the polypeptide may be a BiKE or a CAR-NK. The polypeptide may be a BiKE or a CAR-NK comprising a blockade of a killer cell immunoglobulin-like receptor (KIR), NKG2A, immunoglobulin-like transcript (ILT), or any combination thereof. The polypeptide may be a tri-specific antibody.
In various cases described herein, the peptide conjugate/MHC complex can be presented on a surface of a cell. The cell can express a low copy number of the peptide conjugate/MHC complex, and wherein the low copy number is at most about 1000, 900, 800, 700, 600, 500, 400, 300, 200, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10, 5, 3, or 1 copy per a single cell. The peptide of the peptide conjugate/MHC complex can be from an intracellular protein. For example, the peptide can be processed from an endogenous or an intracellular protein and presented on the MHC complex.
The binding partners described herein can be a T-cell receptor (TCR) or a functional fragment thereof (e.g., antigen-binding fragment, variable region or extracellular domain of a TCR). The binding partners described herein can be a TCR mimetic binder. The T-cell receptor or the functional fragment thereof can bind specifically to the peptide conjugate/MHC complex described herein. In some cases, the KD of the TCR mimetic binder for the peptide conjugate/MHC complex can be less than about IpM, less than about 500 nM, less than about 400 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 1 nM, less than about 0.1 nM, less than about 50 pM, less than about 20 pM, less than about 10 pM, less than about 1 pM, less than about 0.1 pM, or less. In some cases, the KD of the TCR for the peptide conjugate/MHC complex can be less than about IpM, less than about 500 nM, less than about 400 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 1 nM, or less than about 0.1 nM, less than about 50 pM, less than about 20 pM, less than about 10 pM, less than about 1 pM, less than about 0.1 pM, or less.
In an aspect, the present disclosure provides a method of identifying a T-cell receptor (TCR) that recognizes the peptide conjugate/MHC complex described herein. In some embodiments, the method of identifying a TCR comprises contacting a plurality of candidate TCRs with the peptide conjugate/MHC complex and identifying at least one TCR that binds to the peptide conjugate/MHC complex. In some embodiments, the method of identifying a TCR that recognizes the peptide conjugate/MHC complex described herein comprises selecting or isolating at least one TCR. Isolation of a TCR can be achieved through analysis of binding affinities to peptide conjugate/MHC complexes. In some instances, the plurality of candidate TCRs is a plurality of soluble TCRs. In some instances, the plurality of candidate TCRs is a plurality of TCRs expressed on the cell surface of a plurality of cells. In some embodiments, the method of identifying a TCR that recognizes the peptide conjugate/MHC complex described herein comprises isolating or selecting a cell comprising at least one TCR based on an activation marker of the cell. In some embodiments, the activation marker is a T cell marker. In embodiments, the T cell activation marker is CD26, CD27, CD28, CD30, CD154, CD40L, CD134. CD25, CD44, CD69, CD137, PD-1 or KLRG1. In another aspect, the disclosure provides a T-cell receptor (TCR) comprising at least one identified TCR that recognizes the peptide conjugate/MHC complex described herein. In some embodiments, the TCR is a soluble TCR. In embodiments, the TCR is a bispecific TCR. In some embodiments, the TCR is expressed on a CD4+ T cell. In some embodiments, the TCR is expressed on a CD8+ T cell.
In embodiments, the binding partners are multivalent. In embodiments, a tri-specific binding partner is provided. In embodiments, cells express at least a segment of one or more binding partners in the form of a CAR. In an embodiment, a binding partner of this disclosure may be provided as a complex with a polynucleotide, such as an RNA polynucleotide, to form an aptamer. In embodiments, a multivalent binding partner includes one binding component, such as a paratope, that confers specificity to a particular target on a desired cell type, such as any cancer cell marker. In embodiments, a tri-specific leukocyte engager is provided. In embodiments, the binding partners may be part of a molecule that is activated only in the presence of a protease or other enzyme present in a tumor microenvironment, such embodiments being pertinent to, for instance, a probody, examples of which are known in the art, for example in doi: 10.1126/scitranslmed.3006682, doi: 10.1038/s41467-020-16838-w, and doi: 10.1038/s41587-019-0135-x, from which the descriptions of probodies, and protease activation, are incorporated herein by reference. In an embodiment, the disclosure provides a universal hapten that can be grafted onto inhibitors.
In embodiments, a CAR of this disclosure comprises scFv that comprises heavy and light chain variable regions as described herein. As is known in the art for previously described CARs, the scFv is present in a contiguous polypeptide that further comprises a CD3zeta chain and a costimulatory domain. In some embodiments, the contiguous polypeptide further comprises a CD3gamma chain or a CD3epsilon chain. In embodiments, the costimulatory domain comprises a 4-1BB (CD137) costimulatory domain or a CD28 costimulatory domain. A CAR may also contain a co-receptor hinge sequence, such as a CD8 a co-receptor hinge sequence.
In embodiments, binding partners of this disclosure may comprise a constant region, e.g., an Fc region. Any isotype of constant region can be included. Binding partners that comprise a constant region may be particularly adapted for antibody-dependent cell mediated cytotoxicity (ADCC) and thus may function to kill targeted cells by cell-mediated responses by any of a variety of effector cells. Similarly, a constant region may be particularly adapted for enhancing complement-mediated responses.
In embodiments, a binding partner of this disclosure may be modified such that it is present in a fusion protein. In embodiments, an antigen-binding segment of a binding partner may be present in a fusion protein, and/or the constant region may be a component of a fusion protein. In embodiments, a fusion protein comprises amino acids from at least two different proteins. Fusion proteins can be produced using any of a wide variety of standard molecular biology approaches, including but not necessarily limited to expression from any suitable expression vector. In embodiments, a binding partner described herein may be present in a fusion protein with a detectable protein, such as green fluorescent protein (GFP), enhanced GFP (eGFP), mCherry, and the like. In embodiments, as an alternative to an expression vector, an mRNA or chemically modified mRNA encoding any binding partner described herein can be delivered to cells such that the binding partner is translated by the cells. In embodiments, the fusion proteins comprise a binding partner disclosed herein and a cytokine. In embodiments, the cytokine is IL-2, IL-7, IL-15, IL-12, IL-18, or IL-21. In embodiments, the cytokine is modified to increase at least one therapeutic property, including but not limited to bioavailability, efficacy, extension of half-life, or other desirable properties. Examples of suitable cytokine modifications are described in Front. Immunol., 14 October 2021, doi.org/10.3389/fimmu.2021, the disclosure of which is incorporated herein by reference. In embodiments, the cytokine is modified to decrease binding affinity.
In embodiments, binding partners described herein are used to carry drugs or toxins, and thus the binding partners may be provided as immunotoxins, or in the form of antibodydrug conjugates (ADCs). In some embodiments, the binding partner described herein may be linked to an enzyme. In some embodiments, the binding partner described herein may be linked to a sialidase. Conjugation of sialidase to the binding partner may inactivate one or more inhibitory receptors (e.g., 1, 2, 3, 4, 5, or more inhibitory receptors) at once.
In embodiments, agents useful in the generation of immunotoxins include enzymatically active toxins and enzymatically active fragments thereof. Suitable enzymatically active toxins include but are not limited to diphtheria A chain, nonbinding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, alpha sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin and the tricothecenes. These can be provided as components of fusion proteins or can be covalently attached to the binding partner by any suitable conjugation approach.
The binding partner may be connected to a chemotherapeutic agent by using any suitable linker to form an antibody drug conjugate (ADC). In embodiments, the linker comprises a disulfide, a hydrazine, or a thioether. In embodiments, the ADC comprises two payloads (e.g., two different chemotherapeutic agents), each linked to the binding partner by a linker. In embodiments, the ADC comprises two payloads (e.g., two different chemotherapeutic agents), each linked to the binding partner by a linker using site specific aldehyde tags. The chemotherapeutic agent may be reversibly or irreversibly attached to the binding partner.
Cleavable linkers may be particularly useful for killing bystander cells. In embodiments, a protease recognition site may be included to liberate the chemotherapeutic agent from the binding partner by operation of a protease that recognizes and cleaves at the protease recognition site. The ADC may therefore be considered to contain a prodrug.
In embodiments, binding partners of this disclosure may comprise linking sequences. As a non-limiting example, an ScFv may comprise a linker that links segments comprising paratopes to one another. Suitable amino acid linkers may be mainly composed of relatively small, neutral amino acids, such as glycine, serine, and alanine, and can include multiple copies of a sequence enriched in glycine and serine. In specific and non-limiting embodiments, the linker comprises 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 amino acids. In an example, the linker may be the glycine-serine-alanine linker G4SA3 or a glycine-serine linker (G4S)4 linker. In embodiments, a binding partner may include a cellular localization signal, or a secretion signal. In embodiments, binding partner may comprise a transmembrane domain, and thus may be trafficked to, and anchored in a cell membrane. For secretion, any suitable secretion signal can be used, and many are known in the art.
In embodiments, the binding partners can be part of an ADC and therefore the binding partners comprise a drug. The drug can include, but is not necessarily limited to, any suitable chemotherapeutic agent. In embodiments, the ADC comprises a binding partner and a chemotherapeutic agent that is an anti-microtubule agent, an alkylating agent, or a DNA minor groove binding agent. In embodiments, the chemotherapeutic agent comprises a maytansinoid, a dolastatin, an auristatin drug analog, or a cryptophycin. In embodiments, the chemotherapeutic agent is a duocarmycin derivative, or an antibiotic, such as an enediyne antibiotic, or pyrolobenodiazepine (PBD), including dimers thereof. In embodiments, the chemotherapeutic agent is an enzyme inhibitor, such as a topoisomerase or polymerase inhibitor. In embodiments, the chemotherapeutic agent comprises doxorubicin, or a metalcontaining compound, such as a platinum-containing compound, non-limiting examples of which include cisplatin, carboplatin or oxaliplatin. In embodiments, the ADC comprises a binding partner described herein, and any drug that is described in Barf and Kaptein, dx.doi.org/10.1021/jm3003203, J. Med. Chem. 2012, 55, 6243-6262, or in Wilson et al., dx.doi.org/10.1021/jm400224q, J. Med. Chem. 2013, 56, 7463-7476, or Lambert and Morris, Adv Ther (2017) 34: 1015-1035, from which the descriptions of drugs for use as components as ADCs is incorporated herein by reference. In embodiments, the binding partner is conjugated to or otherwise includes a cytokine, including but not necessarily limited to an interleukin, including but not limited to IL-2, IL-7, IL-8, IL-15, IL-12, IL-18, or IL-21, or an interferon (IFN), to thereby provide a cytokine conjugate. In embodiments, the binding partner comprises a toxin conjugate. The toxin conjugate can include, but is not necessarily limited to Pseudomonas exotoxin A (PE), diphtheria toxin (DT), or a recombinant variant thereof. In some embodiments, the binding partner can comprise an immunotoxin agent derived from PE or DT, including but not limited to BL22, LMB-2, CAT-8015, SS1P, MR1- 1, or Zemab.
For production of binding partners, any suitable expression system may be used. In general, polynucleotides encoding binding partners are used to express the binding partners in any suitable cell system, non-limiting embodiments of which include NSO murine myeloma cells, human cell lines, and Chinese hamster ovary (CHO) cells. In embodiments, the disclosure provides a polynucleotide that can selectively hybridize to a polynucleotide encoding any CDR or combination of CDRs described herein. In embodiments, the polynucleotide selectively hybridizes to a polynucleotide encoding a heavy chain CDR1, CDR2, and CDR3 of any described binding partner. In embodiments, the polynucleotide selectively hybridizes to a polynucleotide encoding a light chain CDR1, CDR2, and CDR3 of any described binding partner. In embodiments, the polynucleotide selectively hybridizes to a polynucleotide encoding CDR1, CDR2, and CDR3 of a heavy and light chain of any described binding partner.
In embodiments, a binding partner described herein may be a component of a fusion protein. In embodiments, such as for a binding partner that is produced as a fusion protein, a peptide linker may be used. In embodiments, the peptide linker comprises any self-cleaving signal. In embodiments, the self-cleaving signal may be present in the same open reading frame (ORF) as the ORF that encodes the binding partner. A self-cleaving amino acid sequence is typically about 18-22 amino acids long. Any suitable sequence can be used, nonlimiting examples of which include: T2A (EGRGSLLTCGDVEENPGP); P2A (ATNFSLKQAGDVENPGP); E2A (QCTNYALKLAGDVESNPGP) and F2A (VKQTLNFDLKLAGDVESNPGP).
To the extent any segment of a protein comprising a binding partner described herein was a component of a library, including but not necessarily limited to a phage display library or a yeast surface display library, the disclosure includes the proviso that the binding partner may be free of any segment of the library that comprises a bacteriophage or yeast amino acid sequence, including but not limited to phage coat protein or a yeast host protein, including but not limited to Aga2. Thus, in certain embodiments, the binding partner may be present in a fusion protein, but the fusion protein does not comprise bacteriophage coat protein. In embodiments, any binding partner described herein may be free of any of pill phage coat protein, or any part of M13, fd filamentous phage, T4, T7, or X phage protein.
In embodiments, a binding partner of this disclosure comprises a detectable label, which may be used for diagnostic or therapeutic purposes. For example, a detectable label can be used for localization of the binding partner for pathology and/or in vivo imaging approaches. In embodiments, a binding partner is conjugated to any of a variety of radioactive agents, including but not limited to a highly radioactive atom, such as Ini 11, At211, 1131, 1125, Y90, Rel86, Rel88, Sml53, Bi212, P32, Pb212, and radioactive isotopes of Lu. In particular embodiments, such as for imaging, the binding partner may be conjugated to a radioactive atom for scintigraphic approaches, for example Tc99m (metastable technetium-99), 1123, or a spin label for nuclear magnetic resonance (NMR) imaging (also known as magnetic resonance imaging, or “MRI”), such as 1123, 1131, 1124, F19, C13, N15, 017 or Gadlinium (III) or Manganese (II). In embodiments, the radioactive agent is suitable for use in CAT scan or PET imaging. In embodiments, Indiuml 11, Technetium99 or Iodine 131 can be used for planar scans or single photon emission computed tomography (SPECT). Positron emitting labels such as Fluorinel9 Iodine 123 and Iodine 124 can be used in positron emission tomography. Paramagnetic ions such as Gadlinium (III) or Manganese (II) can used in magnetic resonance imaging MRI. In embodiments, the described radioactive isotopes that are attached to a described binding partner can also be used in therapeutic approaches. In embodiments, radioactive agents or isotopes include alpha-emitting radionuclides. In embodiments, radioactive agents or isotopes include beta-emitting radionuclides. In some embodiments, the present disclosure provides an antibody of the present technology conjugated to a diagnostic or therapeutic agent. The diagnostic agent may comprise a non-radioactive label, a contrast agent (such as for magnetic resonance imaging, computed tomography or ultrasound), and/or a radioactive label which can be a gamma-, beta-, alpha-, Auger electron-, or positron-emitting isotope. A diagnostic agent is a molecule which is administered conjugated to an antibody moiety, i.e., antibody or antibody fragment, or subfragment, and is useful in diagnosing or detecting a disease by locating the cells containing the antigen.
The polypeptide can contact the alpha 1 domain and alpha 2 domain of a heavy chain of the MHC. The polypeptide can bind to an epitope of the MHC. The epitope can comprise one or more residues from the regions comprising residues 62-66, 106-109, and/or 150-170 of the MHC. The polypeptide can bind to an epitope of the MHC, and the epitope can comprise one or more residues from the regions comprising residues 62-66, 106-109, and/or 150-170 of the HLA-A*03:01 or HLA-A* 11:01. The polypeptide can bind to an epitope of the MHC, and the epitope can comprise one or more residues selected from the group consisting of residues 62, 106, 108, 109, 158, 161, 162, 162, 165, 166, 167, 169 and 170 of the HLA-A*03:01. The polypeptide can bind to an epitope of the MHC, and the epitope can comprise one or more residues selected from the group consisting of residues 62, 106, 108, 109, 158, 161, 162, 162, 165, 166, 167, 169 and 170 of the HL A-A* 03:01. The polypeptide can bind to an epitope of the MHC, and the epitope can comprise one or more residues selected from the group consisting of residues 62, 65, 66, 150, 151, 154, 155, 157, and 158 of the HLA-A*03:01. The polypeptide can bind to an epitope of the MHC, and the epitope can comprise one or more residues selected from the group consisting of residues 62, 65, 66, 150, 151, 154, 155, 157, and 158 of the HLA-A*03:01. The polypeptide can bind to an epitope of the MHC, and the epitope can comprise one or more residues selected from the group consisting of residues 62, 106, 108, 109, 154, 157, 158, 161, 162, 163, 165, 166, 167, 169 and 170 of the HLA-A* 11 :01. The polypeptide can bind to an epitope of the MHC, and the epitope can comprise one or more residues selected from the group consisting of residues 62, 106, 108, 109, 154, 157, 158, 161, 162, 163, 165, 166, 167, 169 and 170 ofthe HLA- A* 11 :01. The polypeptide can bind to an epitope of the MHC, and the epitope can comprise one or more residues selected from the group consisting of residues 62, 65, 66, 151, 154, 155 and 158 of the HLA-A* 11 :01. The polypeptide can bind to an epitope of the MHC, and the epitope can comprise one or more residues selected from the group consisting of residues 62, 65, 66, 151, 154, 155 and 158 ofthe HLA-A* 11 :01.
In some embodiments, the targeted covalent inhibitor is (i) a tri-complex KRAS inhibitor or a KRAS degrader, (ii) a tri-complex KRAS inhibitor or a KRAS degrader, or (iii) a tri-complex KRAS inhibitor or a KRAS degrader. In some embodiments, the targeted covalent inhibitor is (i) a tri-complex KRASG12C inhibitor or a KRASG12C degrader, (ii) a tri-complex KRASG12D inhibitor or a KRASG12D degrader, (iii) a tri-complex KRASG12R inhibitor or a KRASG12R degrader, or (iv) a tri-complex KRASG12S inhibitor or a KRASG12S degrader. A tri-complex KRAS inhibitor can form a complex with KRAS, or a mutant thereof, and another molecule. The molecule may be an immunophilin. A KRAS degrader can invoke proteolysis of the KRAS peptide. Proteolysis can occur via ubiquitin-proteosome systems.
In some aspects, the present disclosure provides methods of identifying or designing polypeptides that can bind to a peptide-conjugate/MHC complex based on three-dimensional structure(s) of the peptide-conjugate/MHC complex. The three-dimensional structures of the peptide-conjugate/MHC complex may be obtained from existing complex structures of binding partners binding to the peptide-conjugate/MHC complexes (e.g., the structures provided herein). Computer-assisted methods can be used to identify or design polypeptides that can link to a targeted covalent inhibitor, or fragment thereof, that is covalently linked to the peptide of a peptide conjugate/MHC complex.
In some embodiments of the computer-assisted method for designing polypeptides that can link to a targeted covalent inhibitor, interactions between atoms of the antigenbinding domain and atoms of the peptide are determined. For example, such a method can comprise: (a) providing the structure of a polypeptide comprising an antigen-binding domain bound to a peptide conjugate/MHC complex; (b) determining interactions between one or more atoms of the antigen-binding domain of the polypeptide and one or more atoms of the peptide conjugate/MHC complex; (c) providing a candidate polypeptide comprising an antigen-binding domain for binding to a peptide conjugate/MHC complex, wherein the antigen-binding domain of the candidate polypeptide comprises one or more amino acid substitutions relative to the polypeptide comprising an antigen-binding domain, wherein the one or more amino acid substitutions are of residues of the antigen-binding domain of the polypeptide that comprise the one or more atoms that interact with or modulate interaction with the one or more atoms of the peptide conjugate/MHC complex; and (d) selecting the candidate polypeptide if it binds or is predicted to bind to the peptide conjugate/MHC complex with a higher affinity than the affinity of the polypeptide comprising an antigenbinding domain to the peptide conjugate/MHC complex.
Any binding partner described herein may be fully or partially humanized. Techniques for humanization of antibodies are known in the art and can be adapted for use in the present disclosure. In embodiments, humanization may be performed, for example, by CDR-grafting. In embodiments, for humanization or to otherwise improve a characteristic of the binding partners, one or more amino acids in a variable region can be changed. In embodiments, one or more amino acids in a framework region can be changed.
The disclosure includes binding partners for use in diagnostic and therapeutic approaches. For therapeutic approaches, in certain embodiments, binding partners may be delivered as mRNA or DNA polynucleotides that encode the binding partners. It is considered that administering a DNA or RNA encoding any binding partner described herein is also a method of delivering such binding partners to an individual or one or more cells. The polynucleotide may be a modified polynucleotide. In some embodiments, the mRNA comprises synthetic mRNA containing modified nucleotides. In some embodiments, the modifications can include Nl-methyl-pseudouri dine (ImT) nucleotide substitutions and/or 5- methylcytidine (m5C) substitutions. Methods of delivering DNA and RNAs encoding proteins are known in the art and can be adapted to deliver the binding partners, given the benefit of the present disclosure. In embodiments, one or more expression vectors are used and comprise viral vectors. Thus, in embodiments, a viral expression vector is used. Viral expression vectors may be used as naked polynucleotides, or may comprise any of viral particles, including but not limited to defective interfering particles or other replication defective viral constructs, and virus-like particles. In embodiments, the expression vector comprises a modified viral polynucleotide, such as from an adenovirus, a herpesvirus, or a retrovirus. In embodiments, a retroviral vector adapted from a murine Moloney leukemia virus (MLV) or a lentiviral vector may be used, such as a lentiviral vector adapted from human immunodeficiency virus type 1 (HIV-1). In some embodiments, the polynucleotides encoding the binding partners may be delivered as nanoparticles. In some embodiments, the RNAs (or mRNAs) encoding the binding partners may be delivered as nanoparticles. In some embodiments, the RNAs (or mRNAs) encoding the binding partners may be delivered as lipid nanoparticles. In some embodiments, the RNA encoding the binding partner can be a modified RNA. The modifications can include Nl-methyl-pseudouri dine ( I m ) nucleotide substitutions and/or 5-methylcytidine (m5C) substitutions.
In an embodiment, an oncolytic viral vector is used. Oncolytic viruses (oVs), including vaccinia (OVV), mediate anticancer effects by both direct oncolysis and stimulation of innate immune responses through production of damage-associated molecular patterns (DAMPs) and the presence of virus-derived pathogen-associated molecular patterns (PAMPs), leading to increased type I interferon production. Additionally, OVV-mediated oncolysis may facilitate the direct acquisition of tumor-derived antigens by host antigen-presenting cells within the tumor microenvironment, thereby leading to improved T cell priming as well as coordination of the effector phase of antitumor immune responses. In alternative embodiments, a recombinant adeno-associated virus (AAV) vector may be used. In certain embodiments, the expression vector is a self- complementary adeno-associated virus (scAAV).
Pharmaceutical formulations containing binding partners are included in the disclosure and can be prepared by mixing them with one or more pharmaceutically acceptable carriers. Pharmaceutically acceptable carriers include solvents, dispersion media, isotonic agents, and the like. The carrier can be liquid, semi-solid, e.g. pastes, or solid carriers. Examples of carriers include water, saline solutions or other buffers (such as phosphate, citrate buffers), oil, alcohol, proteins (such as serum albumin, gelatin), carbohydrates (such as monosaccharides, disaccharides, and other carbohydrates including glucose, sucrose, trehalose, mannose, mannitol, sorbitol or dextrins), gel, lipids, liposomes, resins, porous matrices, binders, fillers, coatings, stabilizers, preservatives, liposomes, antioxidants, chelating agents such as EDTA, salt forming counter-ions such as sodium; nonionic surfactants such as TWEEN, PLURONICS or polyethylene glycol (PEG), or combinations thereof. In some embodiments, a liposomal formulation comprising one or more binding partners is provided. Liposomal formulations include but are not limited to liposomal nanoparticles.
In another aspect, the present disclosure provides a pharmaceutical composition comprising a polypeptide disclosed herein, and a pharmaceutically acceptable carrier.
In another aspect, the present disclosure provides a method of treating cancer in a subject that has been treated with a free targeted covalent inhibitor, the method comprising administering to the subject the polypeptide disclosed herein or the pharmaceutical composition disclosed herein. In some embodiments, the subject is refractory to a treatment with the free targeted covalent inhibitor.
In another aspect, the present disclosure provides a method of treating cancer in a subject, the method comprising administering to the subject the polypeptide disclosed herein or the pharmaceutical composition disclosed herein after or simultaneously with administration of a small molecule drug.
In another aspect, the present disclosure provides a method of treating cancer in a subject, the method comprising administering to the subject the targeted covalent inhibitor, and administering to the subject the polypeptide disclosed herein or the pharmaceutical composition disclosed herein. In some embodiments, the polypeptide disclosed herein or the pharmaceutical composition disclosed herein is administered after administration of the targeted covalent inhibitor or simultaneously with the targeted covalent inhibitor.
In embodiments, an effective amount of one or more binding partners is administered to an individual in need thereof. In embodiments, an effective amount is an amount that reduces one or more signs or symptoms of a disease and/or reduces the severity of the disease. An effective amount may also inhibit or prevent the onset of a disease or a disease relapse. A precise dosage can be selected by the individual physician in view of the patient to be treated. Dosage and administration can be adjusted to provide sufficient levels of binding partner to maintain the desired effect. Additional factors that may be taken into account include the severity and type of the disease state, age, weight, and gender of the patient, desired duration of treatment, method of administration, time and frequency of administration, drug combination(s), reaction sensitivities, and/or tolerance/response to therapy.
Binding partners and pharmaceutical compositions comprising the binding partners can be administered to an individual in need thereof using any suitable route, examples of which include intravenous, intramuscular, intraperitoneal, intracerobrospinal, subcutaneous, intra-articular, intrasynovial, oral, topical, or inhalation routes, depending on the particular condition being treated. The compositions may be administered parenterally or enterically. The compositions may be introduced as a single administration or as multiple administrations or may be introduced in a continuous manner over a period of time. For example, the administration(s) can be a pre-specified number of administrations or daily, weekly, or monthly administrations, which may be continuous or intermittent, as may be therapeutically indicated.
In embodiments, the individual in need of a composition of this disclosure has been diagnosed with or is suspected of having cancer. In embodiments, the cancer is a solid tumor or a hematologic malignancy. In embodiments, the cancer is renal cell carcinoma, breast cancer, prostate cancer, pancreatic cancer, lung cancer (e.g. non-small-cell lung cancer), liver cancer, ovarian cancer, cervical cancer, colon cancer, esophageal cancer, glioma, glioblastoma or another brain cancer, stomach cancer, bladder cancer, testicular cancer, head and neck cancer, thyroid cancer, adrenal cancer, endometrial cancer, melanoma or another skin cancer, any sarcoma, including but not limited to fibrosarcoma, angiosarcoma, osteosarcoma, and rhabdomyosarcoma, and any blood cancer, including all types of leukemia, lymphoma, and myeloma. In some embodiments, the individual is in need of treatment for a neuroendocrine tumor. In some embodiments, the individual is in need of treatment for any pre-neoplastic disorder, including myelodysplastic syndromes or myeloproliferative neoplasms. In embodiments, a described binding partner is used prophylactically for any of the described types of cancer. In some embodiments, the individual in need thereof has been treated with a prior treatment and the individual is refractory to the prior treatment. In some embodiments, the cancer is a relapsed or refractory cancer.
In embodiments, administering one or more binding partners, including but not necessarily in a pharmaceutical formulation, to an individual in need thereof, exhibits an improved activity relative to a control. In an embodiment, the control comprises different antibodies, a different form of the same antibodies/binding partner, or antibodies/binding partners that are delivered without adding additional agents. In embodiments, a binding partner described herein provides for improved antibody dependent cell cytotoxicity (ADCC), or for internalization (such as for an ADC), relative to a control. In embodiments, a control protein or peptide does not comprise the covalently linked molecule. The control peptide may comprise the same sequence as the experimental peptide, or if the experimental peptide comprises a mutation the control peptide may comprise the wild type sequence.
A composition of this disclosure, such as a pharmaceutical formulation, can contain only one, or more than one binding partner, and thus combinations of different binding partners are included.
In an aspect, provided herein is a method of treating a disease or disorder in a subject, the method comprising administering to the subject: (a) a targeted covalent inhibitor, and (b) a binding partner disclosed herein. In an embodiment, the disease or disorder is cancer, a fibrotic disease, or an autoimmune disease (e.g., rheumatoid arthritis). In an embodiment, the targeted covalent inhibitor targets KRAS.
In an aspect, provided herein is a method of killing a cancer cell in a subject, the method comprising administering to the subject: (a) a targeted covalent inhibitor, and (b) a binding partner disclosed herein. In an embodiment, the targeted covalent inhibitor targets KRAS.
In an aspect, provided herein is a method of targeting a cell that expresses KRASG12C in a subject that has been treated with a KRASG12C targeted covalent inhibitor, the method comprising administering to the subject a binding partner disclosed herein. In an embodiment, the subject has cancer.
In an aspect, provided herein is a method of treating a disease or disorder in a subject that has been treated with a targeted covalent inhibitor, the method comprising administering to the subject a binding partner disclosed herein. In an embodiment, the disease or disorder is an autoimmune disease or a fibrotic disease.
In an aspect, provided herein is a method of treating cancer in a subject that has been treated with a targeted covalent inhibitor, the method comprising administering to the subject a binding partner disclosed herein. In an embodiment, the targeted covalent inhibitor targets KRAS.
The targeted covalent inhibitor can be a non-peptide molecule described herein. The non-peptide molecule can be administered to a subject in need thereof prior to administering the binding partner described herein. The subject described herein can be a cancer patient. In some cases, the binding partner can be administered to a subject already treated with a nonpeptide molecule previously. In some cases, the binding partner can be administered to a subject simultaneously or after administration of a non-peptide molecule (e.g., the targeted covalent inhibitor or the drug described herein). In some cases, the binding partner can be administered to a subject before administration of a non-peptide molecule.
In some cases, the subject described herein is refractory to a first line of treatment. For example, the subject described herein can be refractory to a first line of treatment which is a chemotherapy. For example, the subject described herein can be refractory to a non-peptide molecule such as AMG-510, MRTX849, and/or GDC6036. In some cases, the subject can be further treated or administered with the binding partner described herein (e.g., antibody or fragment thereof) after relapsed from a prior treatment. In some cases, the binding partners described herein can be used to treat a relapsed or refractory cancer.
In some cases, a subject is prophylactically administered the peptide conjugate described herein prior to the subject developing a cancer, or prior to the subject being administered a drug, or both. The cancer can be due to KRAS mutation, e.g., KRASG12C, KRASG12D, or KRASG12R. In some cases, the method provided herein comprises administering a peptide conjugate to a subject prophylactically, thereby preventing the subject from developing a cancer or preventing the subject from developing a drug resistance. The subject can be a patient who would be receiving the drug described herein.
In some cases, provided herein is a method of preventing a disease or condition in a subject that will receive a drug to treat the disease or condition, the method comprising administering the peptide-drug conjugate prophylactically to a subject as a vaccine to a disease or condition treated with a drug, wherein the subject does not have the disease or condition at the time of administration, and wherein the subject will receive the drug if the subject develops the disease or condition.
In some cases, provided herein is a method of preventing drug resistance in a subject in need thereof, the method comprising administering the peptide-drug conjugate to a subject with the disease or condition, wherein the subject has previously received a drug to treat the disease or condition, and wherein the subject has not developed resistance to the drug prior to the administration of the peptide-drug conjugate, thereby preventing resistance to the drug in the subject.
In some cases, provided herein is a method of preventing drug resistance in a subject in need thereof, the method comprising administering the peptide-drug conjugate to a subject with the disease or condition, wherein the subject has not yet received a drug to treat the disease or condition prior to the administration of the peptide-drug conjugate, thereby preventing resistance to the drug in the subject.
In an aspect, provided herein is a method of enhancing immune recognition of a cell expressing a KRASG12C mutation in a subject that has a cancer that exhibits a KRASG12C, the method comprising administering to the subject: (a) a KRASG12C inhibitor, and (b) a binding partner disclosed herein. In an embodiment, the subject has been previously treated with the KRASG12C inhibitor. In an embodiment, the inhibitor is AMG-510, MRTX849, or GDC6036.
In an embodiment, the cancer is renal cell carcinoma, breast cancer, prostate cancer, pancreatic cancer, lung cancer, liver cancer, ovarian cancer, cervical cancer, colon cancer, esophageal cancer, glioma, glioblastoma, brain cancer, stomach cancer, bladder cancer, testicular cancer, thyroid cancer, adrenal cancer, head and neck cancer, melanoma, skin cancer, sarcoma, fibrosarcoma, angiosarcoma, osteosarcoma, rhabdomyosarcoma, leukemia, lymphoma, myeloma, endometrial cancer, or a neuroendocrine tumor. In an embodiment, the targeted covalent inhibitor is AMG-510, MRTX849, or GDC6036.
In some embodiments, the method disclosed herein further comprises administering an additional therapeutic agent. In an embodiment, the additional therapeutic agent is a conventional chemotherapeutic agent, a modulator of T-cell costimulatory molecules, or an immune checkpoint inhibitor. In an embodiment, the additional therapeutic agent is a chemotherapeutic or an immunomodulator. In an embodiment, the immunomodulator is a checkpoint targeting agent.
In an embodiment, the additional therapeutic agent is a checkpoint targeting agent selected from the group consisting of an antagonist anti-PD-1 antibody, an antagonist anti- PD-L1 antibody, an antagonist anti-PD-L2 antibody, an antagonist anti-CTLA-4 antibody, an antagonist anti-BTLA antibody, an antagonist anti-TREMR antibody, an antagonist anti- TIGIT antibody, an antagonist anti-VISTA antibody, an antagonist anti-TIM-3 antibody, an antagonist anti-LAG-3 antibody, an antagonist anti-CEACAMl antibody, an agonist anti- GITR antibody, an agonist anti-OX40 antibody, and an agonist anti-CD137 antibody, an agonist anti-DR3 antibody, an agonist anti-TNFSF14 antibody, an agonist anti-CD27 antibody, an agonist anti-ICOS antibody, an agonist anti-CD28 antibody. In an embodiment, the additional therapeutic agent is radiotherapy. In an embodiment, the anti-PD-1 antibody is pembrolizumab, nivolumab, cemiplimab, dostarlimab, retifanlimab, or toripalimab. In an embodiment, the anti-PD-Ll antibody is avelumab, atezolizumab, or durvalumab. In an embodiment, the anti-CTLA-4 antibody is ipilimumab or tremelimumab. In an embodiment, the anti-LAG-3 antibody is relatlimab. In an embodiment, the additional therapeutic agent is adoptive immunotherapy.
In an embodiment, the immunomodulator is selected from the group consisting of a cytokine, a modified cytokine, a stem cell growth factor, a lymphotoxin, a hematopoietic factor, a colony stimulating factor (CSF), an interferon (IFN), erythropoietin, thrombopoietin, a lymphotoxin (e.g., a tumor necrosis factor (TNF)), a hematopoietic factor such as interleukin (IL), a colony stimulating factor (e.g., granulocyte-colony stimulating factor (G- CSF), granulocyte macrophage-colony stimulating factor (GM-CSF)), an interferon (e.g., interferons -alpha, -beta, -gamma, -lambda), a stem cell growth factor, and an immunomodulatory drug (IMiD/ CelMoD) (e.g., lenalidomide, pomalidomide, avadomide, iberdomide, and GSPT1 degraders (e.g. CC-90009; MRT-2359)).
In an embodiment, the additional therapeutic agent is a glutaminase inhibitor, such as Telaglenastat (CB839). In an embodiment, the additional therapeutic agent is an arginase inhibitor, such as INCB001158 (numidargistat, CB-280). In an embodiment, the additional therapeutic agent is a high-affinity adenosine 2a receptor (A2aR) inhibitor, and/or low- affinity A2bR inhibitor (e.g., imaradenant, ciforadenant, inupadenant, TT-702 etrumadenant and INCB106385). In an embodiment, the additional therapeutic agent is an anti CD73 antibody or small molecule modulator (e.g., AB680, OP5244). In an embodiment, the additional therapeutic agent is a kynurenine pathway inhibitor, TDO and IDO inhibitors. In some embodiments, the additional therapeutic agent is a a2 adrenergic receptor agonist.
In an embodiment, the additional therapeutic agent is an AHR blocker (e.g., BAY2416964 and IK175). In an embodiment, the additional therapeutic agent is a TREX1 modulator, GTP modulator, ATP modulator, cGAMP modulator, cGAMP modulator, cGAS modulator, ENPP1 modulator, CD73 modulator, CD39 modulator, CD38 modulator, TBK1 modulator, PARP7 modulator, IRF3 modulator, or Type I IFN modulator.
Examples of additional therapeutic agents include, but are not limited to, STING agonists such as cyclic dinucleotides (CDNs), SB 11285, SNX281, GSK3745417; ENPP1 inhibitors such as MV626, SR-8314 , SR-8541 (ref. 118); three-prime repair exonuclease 1 (TREX1) inhibitors; PARP7 inhibitors (e.g., RBN-23972), TLR agonists such as TLR7/TLR8 ligand imiquimod, monophosphoryl lipid A (MPL), TLR4 ligand, TLR7 agonist (e.g., LHC 165); DGKa and DGK^ inhibitors; Diacylglycerol (DAG); CBL-B inhibitors: cell- permeable peptides (e.g., APN431) and CBLB -targeting small interfering RNA (siRNA) e.g., APN401) or CBLB small molecule modulators. Examples of additional therapeutic agents include, but are not limited to, antiandrogens (e.g., Casodex, Flutamide, MDV3100, or ARN-509, Enzaluatide, apalutamide, darolutamid and abiraterone), MEK (e.g. MEK1, MEK2, or MEK1 and MEK2) inhibitors (e.g. XL518, CI-1040, PD035901, selumetinib/AZD6244, GSK1120212/trametinib, GDC- 0973, ARRY-162, ARRY-300, AZD8330, PD0325901, U0126, PD98059, TAK-733, PD318088, AS703026, BAY 869766), BRAF inhibitor (e.g. Vemurafenib), CDK4/6 inhibitors, SHP1 inhibitors, SHP2 inhibitors (e.g. TNO155; RMC-4630), PTPN22 inhibitors, PTPN1 inhibitors, PTPN2 inhibitors, CCR2 inhibitors, CXCR2 inhibitors, TORC1/TORC 2 inhibitors, PI-3K-AKT inhibitors, PARP inhibitors (e.g. Olaparib, Niraparib, Rucaparib), CDK4/6 inhibitors, CDK2 inhibitors, CDK7 inhibitors, TEAD inhibitors, alkylating agents (e.g., cyclophosphamide, ifosfamide, chlorambucil, busulfan, melphalan, mechlorethamine, uramustine, thiotepa, nitrosoureas, nitrogen mustards (e.g., mechloroethamine, cyclophosphamide, chlorambucil, meiphalan), ethylenimine and methylmelamines (e.g., hexamethlymelamine, thiotepa), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine, lomusitne, semustine, streptozocin), triazenes (decarbazine)), anti-metabolites (e.g., 5-azathioprine, leucovorin, capecitabine, fludarabine, gemcitabine, pemetrexed, raltitrexed, folic acid analog (e.g., methotrexate), pyrimidine analogs (e.g., fluorouracil, floxouridine, Cytarabine), purine analogs (e.g., mercaptopurine, thioguanine, pentostatin), etc.), plant alkaloids (e.g., vincristine, vinblastine, vinorelbine, vindesine, podophyllotoxin, paclitaxel, docetaxel, etc.), topoisomerase inhibitors (e.g., irinotecan, topotecan, amsacrine, etoposide (VP 16), etoposide phosphate, teniposide, etc.), antitumor antibiotics (e.g., doxorubicin, adriamycin, daunorubicin, epirubicin, actinomycin, bleomycin, mitomycin, mitoxantrone, plicamycin, etc.), platinum-based compounds (e.g. cisplatin, oxaloplatin, carboplatin), anthracenedione (e.g., mitoxantrone), substituted urea (e.g., hydroxyurea), methyl hydrazine derivative (e.g., procarbazine), adrenocortical suppressant (e.g., mitotane, aminoglutethimide), epipodophyllotoxins (e.g., etoposide), antibiotics (e.g., daunorubicin, doxorubicin, bleomycin), enzymes (e.g., L-asparaginase), inhibitors of mitogen-activated protein kinase signaling (e.g. U0126, PD98059, PD184352, PD0325901, ARRY-142886, SB239063, SP600125, or BAY 43-9006), inhibitors of phosphatidylinositol 3-kinase signaling (e.g. wortmannin or LY294002), mTOR inhibitors, antibodies (e.g., rituxan), MAP4K1 inhibitor (e.g ZYF0033), 5-aza-2'-deoxycytidine, doxorubicin, vincristine, etoposide, gemcitabine, imatinib (Gleevec®), geldanamycin, dasatinib, 17-N-Allylamino-17- Demethoxygeldanamycin (17-AAG), bortezomib, carfilzomide, trastuzumab, anastrozole; angiogenesis inhibitors; antiandrogen, antiestrogen; antisense oligonucleotides; apoptosis gene modulators; apoptosis regulators; arginine deaminase; BCR/ABL antagonists; beta lactam derivatives; bFGF inhibitor; bicalutamide; camptothecin derivatives; casein kinase inhibitors (e.g., ICOS, Silmitasertib); clomifene analogues; cytarabine dacliximab; dexamethasone; estrogen agonists; estrogen antagonists; etanidazole; etoposide phosphate; exemestane; fadrozole; finasteride; fludarabine; fluorodaunorunicin hydrochloride; gadolinium texaphyrin; gallium nitrate; gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam; immunostimulant peptides; insulin-like growth factor- 1 receptor inhibitor; interferon agonists; interferons; interleukins; letrozole; leukemia inhibiting factor; leukocyte alpha interferon; leuprolide+estrogen+progesterone; leuprorelin; matrilysin inhibitors; matrix metalloproteinase inhibitors; MIF inhibitor; mifepristone; mismatched double stranded RNA; monoclonal antibody; mycobacterial cell wall extract; nitric oxide modulators; oxaliplatin; panomifene; pentrozole; phosphatase inhibitors; plasminogen activator inhibitor; platinum complex; platinum compounds; prednisone; proteasome inhibitors; protein A-based immune modulator; protein kinase C inhibitor; protein tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; RAS famesyl protein transferase inhibitors; RAS inhibitors; RAS-GAP inhibitor; ribozymes; signal transduction inhibitors; signal transduction modulators; single chain antigen-binding protein; stem cell inhibitor; stem-cell division inhibitors; stromelysin inhibitors; synthetic glycosaminoglycans; tamoxifen methiodide; telomerase inhibitors; thyroid stimulating hormone; translation inhibitors; tyrosine kinase inhibitors; urokinase receptor antagonists; steroids (e.g., dexamethasone), finasteride, aromatase inhibitors, gonadotropin-releasing hormone agonists (GnRH) such as goserelin or leuprolide, adrenocorticosteroids (e.g., prednisone), progestins (e.g., hydroxyprogesterone caproate, megestrol acetate, medroxyprogesterone acetate), estrogens (e.g., diethlystilbestrol, ethinyl estradiol), antiestrogen (e.g., tamoxifen), androgens (e.g., testosterone propionate, fluoxy me sterone), antiandrogen (e.g., flutamide), immunostimulants (e.g., Bacillus Calmette-Guerin (BCG), levamisole, interleukin-2, alphainterferon, etc.), monoclonal antibodies (e.g., anti-CD20, anti-CD22, anti CD25,anti-CD37, anti-CD38, anti-HER2, anti-CD52, anti-HLA-DR, anti Nectin-4, anti Trop2, anti-Mucl, anti- mesothelin, anti-alpha-folate, anti DLL3, anti-GPRNMB, anti-Glypican3, anti-VEGF, anti- EGFR monoclonal antibodies), immunotoxins (e.g., anti-CD33 monoclonal antibody- calicheamicin conjugate, anti-CD22 monoclonal antibody-pseudomonas exotoxin conjugate, anti-CD30 etc.), radioimmunotherapy (e.g., anti-CD20 monoclonal antibody conjugated to U lin, 90Y, or 1311, etc.), triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, topotecan, itraconazole, vindesine, cerivastatin, vincristine, deoxyadenosine, sertraline, pitavastatin, irinotecan, clofazimine, 5-nonyloxytryptamine, vemurafenib, dabrafenib, erlotinib, gefitinib, EGFR inhibitors, epidermal growth factor receptor (EGFR)- targeted therapy or therapeutic (e.g. gefitinib (Iressa™), erlotinib (Tarceva™), cetuximab (Erbitux™), lapatinib (Tykerb™), panitumumab (Vectibix™), vandetanib (Caprelsa™), afatinib/BIBW2992, CI-1033/canertinib, neratinib/HKI-272, CP-724714, TAK-285, AST- 1306, ARRY334543, ARRY-380, AG-1478, dacomitinib/PF299804, OSI-420/desmethyl erlotinib, AZD8931, AEE788, pelitinib/EKB-569, CUDC-101, WZ8040, WZ4002, WZ3146, AG-490, XL647, PD153035, BMS-599626), sorafenib, imatinib, sunitinib, dasatinib, pyrrolo benzodiazepines (e.g. tomaymycin), carboplatin, CC-1065 and CC-1065 analogs including amino-CBIs, nitrogen mustards (such as chlorambucil and melphalan), dolastatin and dolastatin analogs (including auristatins: e.g., monomethyl auristatin E), anthracycline antibiotics (such as doxorubicin, daunorubicin, etc.), duocarmycins and duocarmycin analogs, enediynes (such as neocarzinostatin and calicheamicins), leptomycin derivatives, maytansinoids and maytansinoid analogs (e.g. mertansine), methotrexate, mitomycin C, taxoids, vinca alkaloids (such as vinblastine and vincristine), epothilones (e.g. epothilone B), camptothecin and its clinical analogs topotecan and irinotecan, or the like; vaccines (e.g., Bacillus Calmette-Guerin (BCG), CSF1R inhibitors (e.g., pexidartinib); other class 3 receptor tyrosine kinases (RTKs), such as KIT, FLT3, and platelet-derived growth factor receptors PDGFRa and PDGFRp.
In embodiments, the disclosure comprises administering to an individual in need thereof one or more binding partners and at least one additional agent to provide an additive effect, or a greater than additive effect such as a synergistic result. In embodiments, the described effect comprises inhibition of cancer growth, inhibition of metastasis, or other beneficial effect. An additive effect or synergistic effect may also be achieved by using a combination of at least two described binding partners.
In an aspect, provided herein is a method of detecting a peptide conjugate/MHC complex in a biological sample, the method comprising contacting the sample with a binding partner disclosed herein, wherein the peptide conjugate/MHC complex comprises a peptide conjugate formed by the covalent reaction of a targeted covalent inhibitor with a peptide. In an embodiment, the peptide is a KRASG12C peptide. In an embodiment, the targeted covalent inhibitor is a KRASG12C inhibitor. In an embodiment, the targeted covalent inhibitor is AMG-510, MRTX849, or GDC6036. In an embodiment, the targeted covalent inhibitor is a covalent degrader of an EGFR family kinase. In an embodiment, the biological sample is blood or serum. In an aspect, provided herein is a method of identifying a cell containing a peptide conjugate/MHC complex provided herein, the method comprising contacting the cell with a binding partner disclosed herein.
Various techniques have been developed for the production of binding partners and are included in the scope of this disclosure. In embodiments, the binding partners are produced by host cells by way of recombinant expression vectors. The present disclosure includes all polynucleotide sequences encoding the amino acid sequences described herein, expression vectors comprising such polynucleotide sequences, and in vitro cell cultures comprising such expression vectors. In embodiments, the cell cultures include prokaryotic cells or eukaryotic cells. In embodiments, the cell cultures are mammalian cells. In embodiments, the cells are CHO cells. In embodiments, the cells are HEK293 cells and their derivatives. Kits comprising the binding partners, and/or cell cultures expressing the binding partners, are provided by this disclosure. In general, the kits comprise one or more sealed containers that contain the binding partners, or cells expressing them. Instructions for using the binding partners for therapeutic and/or diagnostic purposes can be included in the kits.
Cells that are modified to express any described binding partner include but are not necessarily limited to CD4+ T cells, CD8+ T cells, Natural Killer T cells, y5 T cells, neutrophils, and cells that are progenitors of T cells, such as hematopoietic stem cells or other lymphoid progenitor cells, such as immature thymocytes (double-negative CD4-CD8-) cells, or double-positive thymocytes (CD4+CD8+). In some embodiments, the cell is optionally a totipotent, multipotent, or pluripotent stem cell, wherein optionally the stem cell has an induced stem cell phenotype, or wherein the cell is optionally a leukocyte. In embodiments, the cell is a macrophage. In some embodiments, the progenitor cells comprise markers, such as CD34, CD117 (c-kit) and CD90 (Thy-1). In embodiments, the modified cells comprise macrophages. In some embodiments, the modified cells comprise neutrophils. In some embodiments, the modified cell is a neutrophil. The described modified cells may be used therapeutically or prophylactically.
In embodiments, the disclosure provides for generation of a binding partner. This approach comprises providing a plurality of distinct binding partners, exposing the plurality of distinct (e.g., different) binding partners to one or a diversity of peptide conjugates, and selecting binding partners that bind with specificity to the peptide conjugates that contain the covalently conjugated drug or other molecule, but do not bind to the protein or peptide that does not comprise the covalently conjugated drug or other molecule. As described above, this approach can be performed on a manner that either does, or does not, require the amino acid sequence of the protein or peptide to be part of the antigenic determinant. The described approach can be used to select binding partners that are specific for presentation of a peptide conjugate as a component of any MHC complex.
In embodiments, binding partners described herein and as otherwise will be apparent by those skilled in the art, can be used to determine whether or not a particular drug or other molecule forms a covalent interaction with a protein or peptide. Thus, the disclosure provides for exposing protein or peptide substrates to drug candidates and using the binding partners described herein or as identified as described herein to determine whether or not the drug forms a covalent interaction with the pertinent substrate. This determination can be made based on whether or not the binding partner binds to the protein or peptide that has been covalently attached to the drug. This approach can be used in lieu of currently available techniques, such as mass spectroscopy and the like.
In an embodiment, the disclosure comprises a method comprising treating an individual in need of said treatment, wherein the individual is optionally in need of treatment for cancer, the method comprising administering to the individual a covalent drug, and administering to the individual a described binding partner that binds with specificity to a peptide that is covalently linked to the covalent drug (a peptide conjugate) wherein optionally the specificity of the binding partner is for the peptide conjugate in an HLA complex. The drug and the binding partner may be administered concurrently or sequentially. In an embodiment, the drug is administered to the individual before the binding partner to thereby form a peptide conjugate that is present in an HLA context, after which the binding partner is administered and binds to the peptide conjugate in the HLA context, and wherein said binding may kill or inhibit growth of the cells that comprise the peptide conjugate in the HLA complex.
In another embodiment the disclosure comprises generating a neoantigen that comprises a covalently attached drug. The method of generating the neoantigen comprises administering to an individual or to cells in vitro a covalent drug, such that the drug is covalently bound to a peptide to thereby produce a neoangtigen comprising a peptide conjugate. The cells in which the neoantigen is generated may be any type of cancer cells, non-limiting examples of which are described herein. The peptide may be introduced into cells, or may be processed from an intact protein by intracellular protein processing. The covalent drug may initially bind to an intact protein, or the covalent drug may initially bind to a peptide. The method may further comprise identifying the neoantigen that comprises the covalently bound drug. Identifying the neoantigen can also include identifying the neoantigen as a component of a particular peptide conjugate-HLA complex. Thus the particular HLA complex in which the neoantigen is present may also be identified. In an embodiment HLA complexes comprising the neoantigen can be separated from cells, purified to any degree of purity, and the neoantigen amino acid sequence can be determined using any suitable technique, a non-limiting example of which is mass spectrometry.
In another aspect the disclosure provides a method of developing binding partners that bind with specificity to the same peptide conjugate presented by a particular HLA type, or a plurality of HLA types. This approach can be implemented using neoantigens that comprise a covalently attached drug generated as described above. Alternatively, samples from individuals who have received a covalent drug may be obtained and used for screening binding partners. Thus, a method provided by the present disclosure comprises providing isolated peptide-conjugate HLA complexes, or cells expressing an HLA in a complex with a peptide conjugate, and screening binding partners to determine binding partners that bind with specificity to the peptide-conjugate HLA complexes. A plurality of binding partners can be screened, such as by using phage or yeast display libraries. Binding partners can be identified by preferential binding to the peptide-conjugate HLA complexes. The preferential binding can be relative to binding to the same peptide conjugate in the same HLA complex but without the covalent drug, or by preferential binding to the peptide-conjugate HLA complex relative to binding to the free covalent drug, or a combination thereof.
In another aspect, the present disclosure provides a method of identifying and isolating T cells that recognize the particular drug-peptide conjugate/MHC complex. This method can be implemented using peripheral mononuclear blood cells (PBMCs, e.g. lymphocytes and leukocytes) obtained from patients previously treated with a targeted covalent inhibitor. Samples from subjects may be obtained and cell-surface marker techniques well known by those in the art can be used to isolate specific T cells.
With respect to developing binding partners that bind with specificity to a plurality of HLA types, the disclosure includes adapting the procedure described above, but using a plurality of different HLAs comprising the same peptide conjugate to screen binding partners. Using this approach facilitates identification of single binding partners that can bind to the same peptide conjugate when presented in different HLA types. Such binding partners expand members of the patient population who are eligible to be treated with a single identified binding partner. In non-limiting embodiments the disclosure provides for identification of single binding partners that can bind with specificity to the same peptide conjugate that is presented by HLA-A*02:01 and at least one other HLA type. In embodiments the disclosure provides for binding partners that can bind with specificity to the same peptide conjugate presented by a combination of HL A-A* 02:01, HLA-A*03:01 and HLA-A* 11 :01.
In some embodiments, the antigen-binding domain described herein binds to (a) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a MRTX849 or fragment thereof presented by HLA-A*03:01; (b) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to MRTX849 or fragment thereof presented by HLA-A*03:01; or (c) a peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a AMG-510 or fragment thereof presented by HLA-A*02:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain described herein binds to (a) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a MRTX849 or fragment thereof presented by HLA-A*03:01; (b) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01; or (c) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain described herein binds to (a) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to MRXT849 or fragment thereof presented by HLA-A*l l:01; (b) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to MRXT849 or fragment thereof presented by HLA-A* 11 :01; or (c) a peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to AMG-510 or fragment thereof presented by HLA-A*02:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRXT849 or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain described herein binds to (a) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to MRTX849 or fragment thereof presented by HLA- A* 11 :01; (b) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01; or (c) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain described herein binds to (a) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to MRTX849 or fragment thereof presented by HLA-A*l l:01; (b) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to MRTX849 or fragment thereof presented by HLA-A* 11:01; or (c) a peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to AMG-510 or fragment thereof presented by HLA-A*02:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain described herein binds to (a) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to MRTX849 or fragment thereof presented by HLA- A* 11 :01; (b) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01; or (c) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain described herein binds to (a) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to MRTX849 or fragment thereof presented by HLA-A*03:01; (b) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to MRTX849 or fragment thereof presented by HLA-A*03:01; or (c) a peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to AMG-510 or fragment thereof presented by HLA-A*02:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain described herein binds to (a) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to MRTX849 or fragment thereof presented by HLA- A*03:01; (b) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG-5 10 or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01; or (c) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*l l :01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01.
In some embodiments, the antigen-binding domain described herein binds to (a) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a AMG-510 or fragment thereof presented by HLA-A*03:01, a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a MRTX849 or fragment thereof presented by HLA-A*03:01, a third peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a GDC6036 or fragment thereof presented by HLA-A*03:01, or any combination thereof; (b) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01, a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to MRTX849 or fragment thereof presented by HLA-A*03:01, a third peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a GDC6036 or fragment thereof presented by HLA-A*03:01, or any combination thereof; or (c) a peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a AMG-510 or fragment thereof presented by HLA-A*02:01, a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01, a third peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a GDC6036 or fragment thereof presented by HLA-A*02:01, or any combination thereof.
In some embodiments, the antigen-binding domain described herein binds to (a) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a AMG-510 or fragment thereof presented by HLA-A*03:01, a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a MRTX849 or fragment thereof presented by HLA-A*03:01, a third peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a GDC6036 or fragment thereof presented by HLA-A*03:01, or any combination thereof; (b) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01, a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01, a third peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a GDC6036 or fragment thereof presented by HLA-A*02:01, or any combination thereof; or (c) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a AMG-510 or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01, a third peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a GDC6036 or fragment thereof presented by HLA-A*03:01, or any combination thereof.
In some embodiments, the antigen-binding domain described herein binds to (a) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01, a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to MRXT849 or fragment thereof presented by HLA-A* 11 :01, a third peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a GDC6036 or fragment thereof presented by HLA-A* 11 :01, or any combination thereof; (b) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01, a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to MRXT849 or fragment thereof presented by HLA-A* 11 :01, a third peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a GDC6036 or fragment thereof presented by HLA-A* 11 :01, or any combination thereof; or (c) a peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to AMG-510 or fragment thereof presented by HLA-A*02:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRXT849 or fragment thereof presented by HLA-A*02:01, a third peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a GDC6036 or fragment thereof presented by HLA-A*02:01, or any combination thereof.
In some embodiments, the antigen-binding domain described herein binds to (a) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01, a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to MRTX849 or fragment thereof presented by HLA- A* 11 :01, a third peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a GDC6036 or fragment thereof presented by HLA-A*02:01, or any combination thereof; (b) a peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to AMG- 510 or fragment thereof presented by HLA-A*03:01, a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01, a third peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a GDC6036 or fragment thereof presented by HLA-A*02:01, or any combination thereof; or (c) a peptide conjugate/MHC complex comprising VVGACGVGK conjugated to AMG-510 or fragment thereof presented by HLA-A*03:01, a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to MRTX849 or fragment thereof presented by HLA-A*02:01, a third peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a GDC6036 or fragment thereof presented by HLA-A*02:01, or any combination thereof.
In embodiments, an individual who is treated with a described binding partner has a condition that is resistant to treatment with the covalent drug that forms part of a peptide conjugate.
In embodiments, binding partners of this disclosure may be used in any immunological diagnostic test, including but not limited to the imaging approaches described above. In embodiments, one or more binding partners described herein can be used as a component in any form of, for example, enzyme-linked immunosorbent assay (ELISA) assay, including but not limited to a direct ELISA, a sandwich ELISA, a competitive ELISA, and a reverse ELISA. In embodiments, one or more binding partners described herein can also be incorporated into an immunodiagnostic device, such as a microfluidic device, a lateral flow device, and the like. The binding partners may also be used in, for example, Western blots and immunoprecipitation assays.
The binding partners described herein can bind to a peptide conjugate/MHC complex. The peptide conjugate can comprise a residue of the peptide linked to a targeted covalent inhibitor described herein. In some embodiments, the peptide comprises at least two amino acid residues between at least two anchor residues. The anchor residues can assist in the peptide conjugate binding to the MHC to form the peptide conjugate/MHC complex. In some embodiments, the anchoring residues may be modified of the peptide sequence to enable higher affinity binding to an HLA molecule. The modification of the anchoring residue(s) may benefit binding to the HLA, conjugation with the targeted covalent inhibitor, and/or refolding of the peptide conjugate/MHC complex. In some embodiments, the peptide may comprise VVDGAGGVGY, VLVGAGGVGV, VYVGAGGVGL, VVVGAGGVGY with the anchoring residues underlined for each peptide. In some embodiments, the peptide may comprise VVDGACGVGY, VLVGACGVGV, VYVGACGVGL, VVVGACGVGY with the anchoring residues underlined for each peptide. In some embodiments, the anchoring residues of a peptide described herein may be modified as shown in FIG. 4A. In some embodiments, modifying the number of anchoring residues may benefit binding to the HLA, conjugation with the targeted covalent inhibitor, and/or refolding of the peptide conjugate/MHC complex. In some embodiments, modifying one or more N-terminal anchoring residues of the peptide may benefit binding to the HLA, conjugation with the targeted covalent inhibitor, and/or refolding of the peptide conjugate/MHC complex. In some embodiments, modifying one or more C-terminal anchoring residues of the peptide may benefit binding to the HLA, conjugation with the targeted covalent inhibitor, and/or refolding of the peptide conjugate/MHC complex.
In some embodiments, residues between the anchoring residues of the peptide may be modified to enhance binding affinity of the binding partner described herein with the peptide- conjugate/MHC complex. In some embodiments, the targeted covalent inhibitor can covalently link to any residue between the anchor residues. The binding affinities of the binding partner described herein to the peptide conjugate/MHC complex with the targeted covalent inhibitor linked to any position in between the anchor residues can be comparable or no more than 200-fold, 100-fold, 50-fold, 20-fold, 10-fold or less difference. In some embodiments, a peptide comprises VVVGACGVGK or VVGACGVGK. In some embodiments, a peptide may comprise the formula: X7X8X9X10X11X12X13X14X15X16. In some cases, a binding partner may bind to a first peptide-conjugate/MHC complex and a second peptide-conjugate/MHC complex with similar affinity (e.g., a similar KD), in which the targeted covalent inhibitors are covalently linked to different residues of the first peptide- conjugate/MHC complex and the second peptide-conjugate/MHC complex. In some embodiments, a similar affinity may comprise two KD values that differ by at most about 100 nM, at most about 90 nM, at most about 80 nM, at most about 70 nM, at most about 60 nM, at most about 50 nM, at most about 40 nM, at most about 30 nM, at most about 20 nM, at most about 15 nM, at most about 10 nM, at most about 9 nM, at most about 8 nM, at most about 7 nM, at most about 6 nM, at most about 5 nM, at most about 4 nM, at most about 3 nM, at most about 2 nM, at most about 1 nM, or less than about 1 nM. In some embodiments, a binding partner may bind to a peptide conjugate/MHC complex with a KD of at most 100 nM, in which the targeted covalent inhibitor is covalently linked to the residue at X9, X10, Xu, or X12. In some embodiments, a binding partner may bind to a peptide conjugate/MHC complex with a KD of at most 100 nM, in which the targeted covalent inhibitor is covalently linked to the residue at X13, X14, or X15. In some embodiments, a binding partner may bind to a peptide conjugate/MHC complex with a KD of at most 100 nM, in which the targeted covalent inhibitor is covalently linked to the residue at X9 position. In some embodiments, a binding partner may bind to a peptide conjugate/MHC complex with a KD of at most 100 nM, in which the targeted covalent inhibitor is covalently linked to the residue at X10 position. In some embodiments, a binding partner may bind to a peptide conjugate/MHC complex with a KD of at most 100 nM, in which the targeted covalent inhibitor is covalently linked to the residue at X11 position. In some embodiments, a binding partner may bind to a peptide conjugate/MHC complex with a KD of at most 100 nM, in which the targeted covalent inhibitor is covalently linked to the residue at X12 position. In some embodiments, a binding partner may bind to a peptide conjugate/MHC complex with a KD of at most 100 nM, in which the targeted covalent inhibitor is covalently linked to the residue at X13 position. In some embodiments, a binding partner may bind to a peptide conjugate/MHC complex with a KD of at most 100 nM, in which the targeted covalent inhibitor is covalently linked to the residue at X14 position. In some embodiments, a binding partner may bind to a peptide conjugate/MHC complex with a KD of at most 100 nM, in which the targeted covalent inhibitor is covalently linked to the residue at X15 position.
In some embodiments, a binding partner may have a greater affinity for a peptide conjugate/MHC complex, in which the targeted covalent inhibitor is covalently linked to the residue at X9 position compared to an affinity of a binding partner to a peptide conjugate/MHC complex, in which the targeted covalent inhibitor is covalently linked to a different residue position of the peptide. In some embodiments, a binding partner may have a greater affinity for a peptide conjugate/MHC complex, in which the targeted covalent inhibitor is covalently linked to the residue at Xio position compared to an affinity of a binding partner to a peptide conjugate/MHC complex, in which the targeted covalent inhibitor is covalently linked to a different residue position of the peptide. In some embodiments, a binding partner may have a greater affinity for a peptide conjugate/MHC complex, in which the targeted covalent inhibitor is covalently linked to the residue at Xu position compared to an affinity of a binding partner to a peptide conjugate/MHC complex, in which the targeted covalent inhibitor is covalently linked to a different residue position of the peptide. In some embodiments, a binding partner may have a greater affinity for a peptide conjugate/MHC complex, in which the targeted covalent inhibitor is covalently linked to the residue at X12 position compared to an affinity of a binding partner to a peptide conjugate/MHC complex, in which the targeted covalent inhibitor is covalently linked to a different residue position of the peptide. In some embodiments, a binding partner may have a greater affinity for a peptide conjugate/MHC complex, in which the targeted covalent inhibitor is covalently linked to the residue at X13 position compared to an affinity of a binding partner to a peptide conjugate/MHC complex, in which the targeted covalent inhibitor is covalently linked to a different residue position of the peptide.
In some aspects, the present disclosure provides a polypeptide that binds to a first peptide conjugate/MHC complex and a second peptide conjugate/MHC complex. The first peptide conjugate/MHC complex and second peptide conjugate/MHC complex can each comprise a peptide. Each peptide may be a different peptide. Each peptide may be the same peptide. In some cases, the peptide may comprise VVVGACGVGK, VVGACGVGKS, VGACGVGKSA, GACGVGKSAL, VVVGCGGVGK, VVVCAGGVGK, VVCGAGGVGK, VVGCGGVGKS, VVCAGGVGKS, VGAGCVGKSA, VGCGGVGKSA, GAGGCGKSAL, GAGCVGKSAL, AGGVCKSALT, AG-GCGKSALT, or AGCVGKSALT. In some cases, the peptide may comprise VVVGACGVGK, VVGACGVGKS, VGACGVGKSA, GACGVGKSAL, VGAGCVGKSA, GAGCVGKSAL, AGCVGKSALT, or AGCVGKSAL. In some cases, the peptide comprises about one (e.g., 0, 1) amino acid substitution in an amino acid sequence as set forth in VVVGACGVGK, VVGACGVGKS, VGACGVGKSA, GACGVGKSAL, VVVGCGGVGK, VVVCAGGVGK, VVCGAGGVGK, VVGCGGVGKS, VVCAGGVGKS, VGAGCVGKSA, VGCGGVGKSA, GAGGCGKSAL, GAGCVGKSAL, AGGVCKSALT, AG-GCGKSALT, AGCVGKSALT, VVVGACGVGK, VVGACGVGKS,VGACGVGKSA, GACGVGKSAL, VGAGCVGKSA, GAGCVGKSAL, AGCVGKSALT, or AGCVGKSAL.
In some embodiments, the peptide may comprise the formula: X#X#+lX#+2X#+3X#+4(X#+5)X#+6X#+7X#+8X#+9, Or X#X#+lX#+2X#+3(X#+4)X#+5X#+6X#+7X#+8X#+9, or X#X#+iX#+2(X#+3)X#+4X#+sX#+6X#+7X#+8X#+9, or X#X#+i(X#+2)X#+3X#+4X#+sX#+6X#+7X#+8X#+9, or X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9X#+io, or X#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9X#+ioX#+i i, or
X#-iX#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9, or X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9X#+ioX#+i2, or X#-iX#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8, or X#-2X#-lX#X#+lX#+2X#+3X#+4(X#+5)X#+6X#+7, Or X#-3X#-2X#-lX#X#+lX#+2X#+3X#+4(X#+5)X#+6. In some embodiments, the peptide may comprise the formula: X#X#+lX#+2X#+3X#+4(X#+5)X#+6X#+7X#+8X#+9, Or X#X#+lX#+2X#+3(X#+4)X#+5X#+6X#+7X#+8X#+9, or X#X#+iX#+2(X#+3)X#+4X#+sX#+6X#+7X#+8X#+9, or X#X#+i(X#+2)X#+3X#+4X#+sX#+6X#+7X#+8X#+9, or X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9X#+io, or X#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9X#+ioX#+i i, or X#-iX#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9, or X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9X#+ioX#+i2, or X#-iX#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8, or X#-2X#-lX#X#+lX#+2X#+3X#+4(X#+5)X#+6X#+7, Or X#-3X#-2X#-lX#X#+lX#+2X#+3X#+4(X#+5)X#+6, or any combination thereof.
The peptide conjugate of the first peptide conjugate/MHC complex and second peptide conjugate/MHC complex can be formed by a covalent reaction of a targeted covalent inhibitor as described herein and the residue in parenthesis of the formula. In some embodiments, the peptide conjugate described herein may form the covalent reaction of a targeted covalent inhibitor and residue X#, X#+i, X#+2, X#+3, X#+4, X#+s, X#+6, X#+7, X#+8, X#+9, X#+io, X#+n, or X#+i2 of a formula provided herein. The first peptide conjugate/MHC complex and second peptide conjugate/MHC complex can be formed by the first peptide conjugate complexing with an MHC molecule and the second peptide conjugate complexing with an MHC molecule. The MHC molecule may be the same MHC. The MHC molecule may be a different MHC.
In some embodiments, a peptide of formula X#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9 can comprise X7X8X9X10X11X12X13X14X15X16, and the peptide conjugate may form by covalently linking the targeted covalent inhibitor to the X12 residue. In some embodiments, a peptide of formula X#X#+i(X#+2)X#+3X#+4X#+sX#+6X#+7X#+8X#+9 can comprise X7X8X9X10X11X12X13X14X15X16, and the peptide conjugate may form by covalently linking the targeted covalent inhibitor to the X9 residue. In some embodiments, a peptide of formula X#X#+iX#+2(X#+3)X#+4X#+sX#+6X#+7X#+8X#+9 can comprise X7X8X9X10X11X12X13X14X15X16, and the peptide conjugate may form by covalently linking the targeted covalent inhibitor to the X10 residue. In some embodiments, a peptide of formula X#X#+iX#+2X#+3(X#+4) X#+sX#+6X#+7X#+8X#+9 can comprise X7X8X9X10X11X12X13X14X15X16, and the peptide conjugate may form by covalently linking the targeted covalent inhibitor to the X11 residue.
In some embodiments, a peptide of formula X#. iX#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9 can comprise X7X8X9X10X11X12X13X14X15X16, and the peptide conjugate may form by covalently linking the targeted covalent inhibitor to the X13 residue.
In some embodiments, the first peptide conjugate/MHC complex and second peptide conjugate/MHC complex can conjugate with a different residue of the peptide. In some embodiments, a peptide can comprise the formula X7X8X9X10X11X12X13X14X15X16 and (i) a first peptide conjugate is formed by the covalent reaction of a first targeted covalent inhibitor or fragment thereof with residue X9, X10, X11, X12, or X13 of the peptide; and (ii) a second peptide conjugate is formed by the covalent reaction of a second targeted covalent inhibitor or fragment thereof with residue X9, X10, X11, X12, X13, X14, or X15 of the peptide.
In some embodiments, a peptide can comprise a formula A-B-C. In some embodiments, A comprises no more than about 5 residues, no more than about 4 residues, no more than about 3 residues, no more than about 2 residues, or no more than about 1 residue. In formula A-B-C, A may comprise an N-terminal anchor residue. The anchor residue may assist with MHC binding of the peptide conjugate. In some embodiments, B comprises at least about 2 residues, at least about 3 residues, at least about 4 residues, at least about 5 residues, at least about 6 residues, at least about 7 residues, at least about 8 residues, at least about 9 residues, or at least about 10 residues. In some embodiments, B comprises no more than 10 residues, no more than 9 residues, no more than 8 residues, no more than 7 residues, no more than 6 residues, no more than 5 residues, no more than 4 residues, no more than 3 residues, or no more than 2 residues. In some embodiments, C comprises no more than about 5 residues, no more than about 4 residues, no more than about 3 residues, no more than about 2 residues, or no more than about 1 residue. In formula A-B-C, C may comprise an C- terminal anchor residue. In some embodiments, a peptide of formula A-B-C can comprise X7X8X9X10X11X12X13X14X15X16 and A may comprise X7, Xs, or X9 or any combination thereof. In some embodiments, A can comprise X7X8. In some embodiments, a peptide of formula A-B-C can comprise X7X8X9X10X11X12X13X14X15X16 and B may comprise X9, X10, Xu, X12, X13, X14, or X15, or any combination thereof. In some embodiments, B can comprise X9X10X11X12X13X14X15. In some embodiments, a peptide of formula A-B-C can comprise X7X8X9X10X11X12X13X14X15X16 and C may comprise X14, X15, or Xi6, or any combination thereof. In some embodiments, C can comprise Xi6.
In some embodiments, a peptide can comprise the formula X7X8X9X10X11X12X13X14X15X16 and the peptide may have a mutation. The mutation may be a G12C mutation at X12 and/or the mutation may be a G13C mutation at X13. In some embodiments, the first peptide conjugate/MHC complex may have a cysteine mutation at X12 (e.g., G12C) covalently linked to the targeted covalent inhibitor (e.g., sotorasib, adagrasib, or divarasib). In some embodiments, the second peptide conjugate/MHC complex may have a cysteine mutation at X12 (e.g., G12C) covalently linked to the targeted covalent inhibitor or fragment thereof (e.g., sotorasib, adagrasib, or divarasib). In some embodiments, the first peptide conjugate/MHC complex may have a cysteine mutation at X13 (e.g., G13C) covalently linked to the targeted covalent inhibitor (e.g., sotorasib, adagrasib, or divarasib). In some embodiments, the second peptide conjugate/MHC complex may have a cysteine mutation at X13 (e.g., G13C) covalently linked to the targeted covalent inhibitor or fragment thereof (e.g., sotorasib, adagrasib, or divarasib).
In some embodiments, a polypeptide described herein may bind with a higher KD to a peptide conjugate/MHC complex with a targeted covalent inhibitor conjugated to a peptide at position X13 compared to a peptide conjugate/MHC complex with a targeted covalent inhibitor conjugated to a peptide at position X12. In some embodiments, the KD may be at least about, at most about, or about 2 fold higher, 3 fold higher, 4 fold higher, 5 fold higher, 6 fold higher, 7 fold higher, 8 fold higher, 9 fold higher, 10 fold higher, 12 fold higher, 15 fold higher, 18 fold higher, or 20 fold higher.
In some embodiments, a polypeptide described herein may bind to a first peptide conjugate/MHC complex and/or a second peptide conjugate/MHC complex with a dissociation constant (e.g., KD) of at most about 200 nM, at most about 150 nM, at most about 100 nM, at most about 90 nM, at most about 80 nM, at most about 70 nM, at most about 60 nM, at most about 50 nM, at most about 40 nM, at most about 30 nM, at most about 20 nM, at most about 10 nM, at most about 1 nM, at most about 100 pM, at most about 50 pM, at most about 10 pM, at most about 1 pM, at most about 100 fM, at most about 50 fM, at most about 10 fM, at most about 1 f , or less than about 1 fM.
In some embodiments, a measure of binding may be a maximum binding signal (e.g., a Bmax value). The Bmax value can refer to a maximum signal detected at equilibrium, measured by a wavelength shift as measured by biolayer interferometry. The biolayer interferometry analysis can comprise taking measurements with a biolayer interferometer at an association period and/or a dissociation period. The biolayer interferometry analysis can comprise taking measurements with a biolayer interferometer at an association period of about 100 seconds, 200 seconds, 300 seconds, 400 seconds, 500 seconds, 600 seconds, 700 seconds, 800 seconds, 900 seconds, 1000 seconds, 1200 seconds, 1500 seconds, 1800 seconds, or 2000 seconds. The biolayer interferometry analysis can comprise taking measurements with a biolayer interferometer at an association period between about 100 seconds, 200 seconds, 300 seconds, 400 seconds, 500 seconds, 600 seconds, 700 seconds, 800 seconds, 900 seconds, 1000 seconds, 1200 seconds, 1500 seconds, 1800 seconds, or 2000 seconds. The biolayer interferometry analysis can comprise taking measurements with a biolayer interferometer at a dissociation period of about 100 seconds, 200 seconds, 300 seconds, 400 seconds, 500 seconds, 600 seconds, 700 seconds, 800 seconds, 900 seconds, 1000 seconds, 1200 seconds, 1500 seconds, 1800 seconds, or 2000 seconds. The biolayer interferometry analysis can comprise taking measurements with a biolayer interferometer at a dissociation period between about 100 seconds, 200 seconds, 300 seconds, 400 seconds, 500 seconds, 600 seconds, 700 seconds, 800 seconds, 900 seconds, 1000 seconds, 1200 seconds, 1500 seconds, 1800 seconds, or 2000 seconds. The biolayer interferometry analysis can comprise taking measurements with a biolayer interferometer at temperature or temperature range. In some embodiments, the biolayer interferometry analysis comprise taking measurements with a biolayer interferometer at temperature of at least about, at most about, or about 20 °C, 25 °C, 30 °C, 31 °C, 32 °C, 33 °C, 34 °C, 35 °C, 36 °C, 37 °C, 38 °C, 39 °C, 40 °C, 45 °C, 50 °C, or any value between a range of any two of these values.
In some embodiments, the polypeptide described herein may bind to a first peptide conjugate/MHC complex and/or a second peptide conjugate/MHC complex with a Bmax that is at least about 0.01 nm, at least about 0.05 nm, at least about 0.1 nm, at least about 0.15 nm, at least about 0.2 nm, at least about 0.25 nm, at least about 0.3 nm, at least about 0.35 nm, at least about 0.4 nm, at least about 0.45 nm, at least about 0.5 nm, at least about 0.6 nm, at least about 0.7 nm, at least about 0.8 nm, at least about 0.9 nm, or greater than about 0.9 nm. In some embodiments, the polypeptide described herein may bind to a first peptide conjugate/MHC complex and/or a second peptide conjugate/MHC complex with a Bmax that is at most about 0.9 nm, at most about 0.8 nm, at most about 0.7 nm, at most about 0.6 nm, at most about 0.5 nm, at most about 0.45 nm, at most about 0.4 nm, at most about 0.35 nm, at most about 0.3 nm, at most about 0.25 nm, at most about 0.2 nm, at most about 0.15 nm, at most about 0.1 nm, at most about 0.05 nm, at most about 0.01 nm, or less than about 0.01 nm.
In some embodiments, the polypeptide described herein may bind to a first peptide conjugate/MHC complex and/or a second peptide conjugate/MHC complex with a Bmax from about 0.1 nm to about 0.9 nm. In some embodiments, the polypeptide described herein may bind to a first peptide conjugate/MHC complex and/or a second peptide conjugate/MHC complex with a Bmax from about 0.1 nm to about 0.2 nm, about 0.1 nm to about 0.3 nm, about 0.1 nm to about 0.4 nm, about 0.1 nm to about 0.5 nm, about 0.1 nm to about 0.55 nm, about 0.1 nm to about 0.6 nm, about 0.1 nm to about 0.65 nm, about 0.1 nm to about 0.7 nm, about 0.1 nm to about 0.75 nm, about 0.1 nm to about 0.8 nm, about 0.1 nm to about 0.9 nm, about 0.2 nm to about 0.3 nm, about 0.2 nm to about 0.4 nm, about 0.2 nm to about 0.5 nm, about 0.2 nm to about 0.55 nm, about 0.2 nm to about 0.6 nm, about 0.2 nm to about 0.65 nm, about 0.2 nm to about 0.7 nm, about 0.2 nm to about 0.75 nm, about 0.2 nm to about 0.8 nm, about 0.2 nm to about 0.9 nm, about 0.3 nm to about 0.4 nm, about 0.3 nm to about 0.5 nm, about 0.3 nm to about 0.55 nm, about 0.3 nm to about 0.6 nm, about 0.3 nm to about 0.65 nm, about 0.3 nm to about 0.7 nm, about 0.3 nm to about 0.75 nm, about 0.3 nm to about 0.8 nm, about 0.3 nm to about 0.9 nm, about 0.4 nm to about 0.5 nm, about 0.4 nm to about 0.55 nm, about 0.4 nm to about 0.6 nm, about 0.4 nm to about 0.65 nm, about 0.4 nm to about 0.7 nm, about 0.4 nm to about 0.75 nm, about 0.4 nm to about 0.8 nm, about 0.4 nm to about 0.9 nm, about 0.5 nm to about 0.55 nm, about 0.5 nm to about 0.6 nm, about 0.5 nm to about 0.65 nm, about 0.5 nm to about 0.7 nm, about 0.5 nm to about 0.75 nm, about 0.5 nm to about 0.8 nm, about 0.5 nm to about 0.9 nm, about 0.55 nm to about 0.6 nm, about 0.55 nm to about 0.65 nm, about 0.55 nm to about 0.7 nm, about 0.55 nm to about 0.75 nm, about 0.55 nm to about 0.8 nm, about 0.55 nm to about 0.9 nm, about 0.6 nm to about 0.65 nm, about 0.6 nm to about 0.7 nm, about 0.6 nm to about 0.75 nm, about 0.6 nm to about 0.8 nm, about 0.6 nm to about 0.9 nm, about 0.65 nm to about 0.7 nm, about 0.65 nm to about 0.75 nm, about 0.65 nm to about 0.8 nm, about 0.65 nm to about 0.9 nm, about 0.7 nm to about 0.75 nm, about 0.7 nm to about 0.8 nm, about 0.7 nm to about 0.9 nm, about 0.75 nm to about 0.8 nm, about 0.75 nm to about 0.9 nm, or about 0.8 nm to about 0.9 nm.
In some embodiments, a binding partner (e.g., polypeptide) described herein may bind to a first peptide conjugate/MHC complex with a greater affinity than to a second peptide conjugate/MHC complex. In some embodiments, a first peptide conjugate of the first peptide conjugate/MHC complex can have a targeted covalent inhibitor covalently linked to a peptide comprising VVVGACGVGK. In some embodiments, a second peptide conjugate of the second peptide conjugate/MHC complex can have a targeted covalent inhibitor covalently linked to a peptide comprising VVVGACGVGK. In some embodiments, the peptide comprises the formula: X7X8X9X10X11X12X13X14X15X16. The first peptide conjugate can comprise the targeted covalent inhibitor covalently linked to X12 of the peptide. The second peptide conjugate can comprise the targeted covalent inhibitor covalently linked to X13 of the peptide.
In some embodiments, a binding partner described herein may bind to the second peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X13 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G13C of a KRAS peptide) with a dissociation constant (KD) that can be at least about 2-fold higher, at least about 3 -fold higher, at least about 4-fold higher, at least about 5-fold higher, at least about 6-fold higher, at least about 7-fold higher, at least about 8-fold higher, at least about 9-fold higher, at least about 10-fold higher, at least about 15-fold higher, at least about 20-fold higher, at least about 50-fold higher, at least about 75-fold higher, at least about 100-fold higher, at least about 125-fold higher, at least about 150-fold higher, at least about 200-fold higher, at least about 500-fold higher, at least about 1,000-fold higher, at least about 2,500-fold higher, at least about 5,000-fold higher, at least about 10,000-fold higher, at least about 25,000-fold higher, at least about 50,000-fold higher, at least about 100,000-fold higher, at least about 150,000-fold higher, at least about 200,000- fold higher, or greater than about 200,000-fold higher than a KD of the binding partner bound to the first peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X12 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G12C of a KRAS peptide). In some embodiments, a binding partner described herein may bind to the second peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X13 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G13C of a KRAS peptide) with a dissociation constant (KD) that can be at most about 200,000-fold higher, at most about 150,000-fold higher, at most about 100,000-fold higher, at most about 50,000-fold higher, at most about 25,000-fold higher, at most about 10,000-fold higher, at most about 5,000-fold higher, at most about 1,000-fold higher, at most about 500-fold higher, 200-fold higher, at most about 175-fold higher, at most about 150-fold higher, at most about 125-fold higher, at most about 100-fold higher, at most about 75-fold higher, at most about 50-fold higher, at most about 20-fold higher, at most about 15-fold higher, at most about 10-fold higher, at most about 9-fold higher, at most about 8-fold higher, at most about 7-fold higher, at most about 6-fold higher, at most about 5-fold higher, at most about 4-fold higher, at most about 3-fold higher, at most about 2-fold higher, or less than about 2-fold higher than a KD of the binding partner bound to the first peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X12 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G12C of a KRAS peptide). In some embodiments, a binding partner described herein may bind to the second peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X13 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G13C of a KRAS peptide) with a dissociation constant (KD) that can be about 2-fold higher to about 20-fold higher than a KD of the binding partner bound to the first peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X12 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G12C of a KRAS peptide). In some embodiments, a binding partner described herein may bind to the second peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X13 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G13C of a KRAS peptide) with a dissociation constant (KD) that can be about 2-fold higher to about 3 -fold higher, about 2-fold higher to about 4-fold higher, about 2-fold higher to about 5-fold higher, about 2-fold higher to about 6-fold higher, about 2-fold higher to about 7-fold higher, about 2-fold higher to about 8-fold higher, about 2-fold higher to about 9-fold higher, about 2-fold higher to about 10-fold higher, about 2-fold higher to about 12-fold higher, about 2-fold higher to about 15-fold higher, about 2-fold higher to about 20-fold higher, about 3 -fold higher to about 4-fold higher, about 3-fold higher to about 5-fold higher, about 3-fold higher to about 6-fold higher, about 3-fold higher to about 7-fold higher, about 3-fold higher to about 8-fold higher, about 3 -fold higher to about 9-fold higher, about 3 -fold higher to about 10-fold higher, about 3 -fold higher to about 12-fold higher, about 3-fold higher to about 15-fold higher, about 3-fold higher to about 20-fold higher, about 4-fold higher to about 5-fold higher, about 4-fold higher to about 6-fold higher, about 4-fold higher to about 7-fold higher, about 4-fold higher to about 8-fold higher, about 4-fold higher to about 9-fold higher, about 4-fold higher to about 10-fold higher, about 4-fold higher to about 12-fold higher, about 4-fold higher to about 15- fold higher, about 4-fold higher to about 20-fold higher, about 5-fold higher to about 6-fold higher, about 5-fold higher to about 7-fold higher, about 5-fold higher to about 8-fold higher, about 5-fold higher to about 9-fold higher, about 5-fold higher to about 10-fold higher, about 5-fold higher to about 12-fold higher, about 5-fold higher to about 15-fold higher, about 5- fold higher to about 20-fold higher, about 6-fold higher to about 7-fold higher, about 6-fold higher to about 8-fold higher, about 6-fold higher to about 9-fold higher, about 6-fold higher to about 10-fold higher, about 6-fold higher to about 12-fold higher, about 6-fold higher to about 15-fold higher, about 6-fold higher to about 20-fold higher, about 7-fold higher to about 8-fold higher, about 7-fold higher to about 9-fold higher, about 7-fold higher to about 10-fold higher, about 7-fold higher to about 12-fold higher, about 7-fold higher to about 15- fold higher, about 7-fold higher to about 20-fold higher, about 8-fold higher to about 9-fold higher, about 8-fold higher to about 10-fold higher, about 8-fold higher to about 12-fold higher, about 8-fold higher to about 15-fold higher, about 8-fold higher to about 20-fold higher, about 9-fold higher to about 10-fold higher, about 9-fold higher to about 12-fold higher, about 9-fold higher to about 15-fold higher, about 9-fold higher to about 20-fold higher, about 10-fold higher to about 12-fold higher, about 10-fold higher to about 15-fold higher, about 10-fold higher to about 20-fold higher, about 12-fold higher to about 15-fold higher, about 12-fold higher to about 20-fold higher, or about 15-fold higher to about 20-fold higher than a KD of the binding partner bound to the first peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X12 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G12C of a KRAS peptide).
In some embodiments, a binding partner described herein may bind to the second peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X13 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G13C of a KRAS peptide) with a dissociation constant (KD) that can be from about 50-fold higher to about 200,000-fold higher than a KD of the binding partner bound to the first peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X12 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G12C of a KRAS peptide). In some embodiments, a binding partner described herein may bind to the second peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X13 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G13C of a KRAS peptide) with a dissociation constant (KD) that can be from about 50-fold higher to about 100-fold higher, about 50-fold higher to about 250-fold higher, about 50-fold higher to about 500-fold higher, about 50-fold higher to about 1,000-fold higher, about 50-fold higher to about 2,500-fold higher, about 50-fold higher to about 5,000-fold higher, about 50-fold higher to about 10,000-fold higher, about 50-fold higher to about 50,000-fold higher, about 50-fold higher to about 100,000-fold higher, about 50-fold higher to about 150,000-fold higher, about 50-fold higher to about 200,000-fold higher, about 100-fold higher to about 250-fold higher, about 100-fold higher to about 500- fold higher, about 100-fold higher to about 1,000-fold higher, about 100-fold higher to about 2,500-fold higher, about 100-fold higher to about 5,000-fold higher, about 100-fold higher to about 10,000-fold higher, about 100-fold higher to about 50,000-fold higher, about 100-fold higher to about 100,000-fold higher, about 100-fold higher to about 150,000-fold higher, about 100-fold higher to about 200,000-fold higher, about 250-fold higher to about 500-fold higher, about 250-fold higher to about 1,000-fold higher, about 250-fold higher to about 2,500-fold higher, about 250-fold higher to about 5,000-fold higher, about 250-fold higher to about 10,000-fold higher, about 250-fold higher to about 50,000-fold higher, about 250-fold higher to about 100,000-fold higher, about 250-fold higher to about 150,000-fold higher, about 250-fold higher to about 200,000-fold higher, about 500-fold higher to about 1,000- fold higher, about 500-fold higher to about 2,500-fold higher, about 500-fold higher to about 5,000-fold higher, about 500-fold higher to about 10,000-fold higher, about 500-fold higher to about 50,000-fold higher, about 500-fold higher to about 100,000-fold higher, about 500- fold higher to about 150,000-fold higher, about 500-fold higher to about 200,000-fold higher, about 1,000-fold higher to about 2,500-fold higher, about 1,000-fold higher to about 5,000- fold higher, about 1,000-fold higher to about 10,000-fold higher, about 1,000-fold higher to about 50,000-fold higher, about 1,000-fold higher to about 100,000-fold higher, about 1,000- fold higher to about 150,000-fold higher, about 1,000-fold higher to about 200,000-fold higher, about 2,500-fold higher to about 5,000-fold higher, about 2,500-fold higher to about 10,000-fold higher, about 2,500-fold higher to about 50,000-fold higher, about 2,500-fold higher to about 100,000-fold higher, about 2,500-fold higher to about 150,000-fold higher, about 2,500-fold higher to about 200,000-fold higher, about 5,000-fold higher to about 10,000-fold higher, about 5,000-fold higher to about 50,000-fold higher, about 5,000-fold higher to about 100,000-fold higher, about 5,000-fold higher to about 150,000-fold higher, about 5,000-fold higher to about 200,000-fold higher, about 10,000-fold higher to about 50,000-fold higher, about 10,000-fold higher to about 100,000-fold higher, about 10,000-fold higher to about 150,000-fold higher, about 10,000-fold higher to about 200,000-fold higher, about 50,000-fold higher to about 100,000-fold higher, about 50,000-fold higher to about 150,000-fold higher, about 50,000-fold higher to about 200,000-fold higher, about 100,000- fold higher to about 150,000-fold higher, about 100,000-fold higher to about 200,000-fold higher, or about 150,000-fold higher to about 200,000-fold higher than a KD of the binding partner bound to the first peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X12 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G12C of a KRAS peptide).
In some embodiments, the targeted covalent inhibitor may bind to a peptide that is a RAS peptide. The RAS peptide may comprise one or more mutations. The mutation may be at a position G12 and/or G13 of the RAS peptide. The mutation may comprise a G12C, G13C, G12R, G12D, or any combination thereof. In some embodiments, the RAS peptide can comprise an amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV. In some embodiments, the RAS peptide can comprise an amino acid sequence of VVVGAGC VGK, VVGAGCVGK, or KLVVVGAGCV. The binding partner described herein can bind to a peptide conjugate complexed with different HLAs. In some embodiments, the peptide-conjugate/MHC complex comprises an MHC encoded by an HLA allele comprising HLA-A*03:01, HLA-A* 11 :01, HLA-A*02:01, HLA-A*68:01, HLA- A*31:01, HLA-A*30:01, HLA-A*33:03, HLA-A*33:01, HLA-A*74:01, HLA-A*34:02, HLA-A*66:01, HLA-A*68:02, HLA-A*02:05, HLA-A*02:02, HLA-A*02:06, or any combination thereof.
In some embodiments, a polypeptide can bind to a first peptide-conjugate/MHC complex (e.g., a peptide-conjugate/MHC complex with a cysteine of the G12C mutation covalently linked to the targeted covalent inhibitor or fragment thereof) with a KD of at least about, at most about, or about, 200 nM, 150 nM, 100 nM, 90 nM, 80 nM, 70 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 1 nM, 100 pM, 50 pM, 10 pM, 1 pM, 100 fM, 50 fM, 10 fM, 1 fM, 1 fM, or a range between any of these two values. In some embodiments, a polypeptide can bind to a second peptide-conjugate/MHC complex (e.g., a peptide-conjugate/MHC complex with a cysteine of the G13C mutation covalently linked to the targeted covalent inhibitor or fragment thereof) with a KD of at least about, at most about, or about, 200 nM, 150 nM, 100 nM, 90 nM, 80 nM, 70 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 1 nM, 100 pM, 50 pM, 10 pM, 1 pM, 100 04, 50 04, 10 04, 1 fM, 1 fM, or a range between any of these two values.
In some embodiments, a polypeptide can contact a first peptide-conjugate/MHC complex and/or a second peptide-conjugate/MHC complex described herein and a maximum wavelength shift may be observed. The maximum wavelength shift can be at least about, at most about, or about, 0.001 nm, 0.005 nm, 0.01 nm, 0.05 nm, 0.1 nm, 0.15 nm, 0.2 nm, 0.25 nm, 0.3 nm, 0.35 nm, 0.4 nm, 0.45 nm, 0.5 nm, 0.6 nm, 0.7 nm, 0.8 nm, 0.9 nm, or a range between any of these two values, wherein the wavelength shift is observed according to a biolayer interferometry analysis.
In some embodiments, a polypeptide (e.g., binding partner) may contact a first peptide-conjugate/MHC complex and/or a second peptide-conjugate/MHC complex described herein and a maximum binding signal may be observed. In some cases, the maximum binding signal may be compared to a reference binding signal, wherein the reference binding signal can be the maximum binding signal of the polypeptide (e.g., multivalent polypeptide) binding to X#X#+iX#+2X#+3X#+4(X#^5)X#+6X#+7X#+8X#+9. In some cases, the polypeptide (e.g., binding partner) contacting the first peptide-conjugate/MHC complex and/or the second peptide-conjugate/MHC complex described herein can produce a maximum binding signal that can be at least about, at most about, or about 0.001-fold, 0.005- fold, 0.01-fold, 0.05-fold, 0.1-fold, 0.5-fold, 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, or 15 -fold of the reference binding signal.
In some embodiments, a maximum wavelength shift of a binding partner binding to the second peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X13 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G13C of a KRAS peptide) can be at least about 2-fold less, at least about 3-fold less, at least about 4-fold less, at least about 5-fold less, at least about 6-fold less, at least about 7-fold less, at least about 8-fold less, at least about 9- fold less, at least about 10-fold less, at least about 15-fold less, at least about 20-fold less, or greater than about 20-fold less than a maximum wavelength shift of the binding partner binding to the first peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X12 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G12C of a KRAS peptide).
In some embodiments, a maximum binding signal of a binding partner binding to the second peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X13 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G13C of a KRAS peptide) can be at least about 2-fold less, at least about 3-fold less, at least about 4-fold less, at least about 5-fold less, at least about 6-fold less, at least about 7-fold less, at least about 8-fold less, at least about 9-fold less, at least about 10-fold less, at least about 15-fold less, at least about 20-fold less, or greater than about 20-fold less than a maximum binding signal of the binding partner binding to the first peptide conjugate/MHC complex (e.g., peptide conjugate with the targeted covalent inhibitor covalently linked to X12 of the peptide, or peptide conjugate with the targeted covalent inhibitor covalently linked to G12C of a KRAS peptide).
In some embodiments, the binder that binds to a peptide conjugate/MHC complex as described herein may comprise R023. In some embodiments, the binding partner comprises the HC CDR1, HC CDR2, and HC CDR3 of a heavy chain variable region amino acid sequence and/or the LC CDR1, LC CDR2, and LC CDR3 of a light chain variable region of a binding partner R023. The amino acid sequence of the light chain variable region (VL) of R023 can comprise: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGV PSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV (SEQ ID NO: 259). The amino acid sequence of the heavy chain variable region (VH) of R023 can comprise: EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYSIHWVRQAPGKGLEWVASISSSSGST SYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGWIAAMDYWGQGTL VTVSS (SEQ ID NO: 260).
In some embodiments, an amino acid sequence of HC CDR1 of binding partner R023 comprises DYSIH (SEQ ID NO: 261). In some embodiments, an amino acid sequence of HC CDR2 of binding partner R023 comprises SISSSSGSTSYADSVKG (SEQ ID NO: 262). In some embodiments, an amino acid sequence of HC CDR3 of binding partner R023 comprises GGWIAAMDY (SEQ ID NO: 263). In some embodiments, an amino acid sequence of LC CDR1 of binding partner R023 comprises RASQSVSSAVA (SEQ ID NO: 264). In some embodiments, an amino acid sequence of LC CDR2 of binding partner R023 comprises SASSLYS (SEQ ID NO: 265). In some embodiments, an amino acid sequence of LC CDR3 of binding partner R023 comprises QQASYVRKTIT (SEQ ID NO: 266).
The following Examples are intended to illustrate but not limit the disclosure. Differences between binding partners may be apparent from the Examples and their accompanying figures. The different properties include, but are not necessarily limited to, specificity for a drug conjugate displayed in the context of a specific MHC type. Thus, binding partners may exhibit different binding partners when a peptide conjugate is in a particular MHC complex. EXAMPLE 1
This Example provides a description of the binding screening and characterization of binding partners that bind with specificity to peptide conjugate/MHC complexes comprising MRTX849.
Clones were identified through selections on a human synthetic antibody phagedisplay library that preferentially bound to peptide conjugate/MHC complexes. Phage- displayed Fab clones were transferred into a yeast display vector in the scFv format. Binding of scFv to biotinylated pMHC complexes (e.g., ada-p?/A03, where ada references the drug adagrasib, p? references the peptide, and A03 references the HLA allele) was tested using yeast display.
As shown in FIG. 1, exemplary binding partners RM_001 and RM_003 showed high binding affinity and high selectivity for ada-p?/A03 complex. Exemplary binding partners, RM_005 and RM_010 showed high binding affinity and selectivity for ada-p?/Al 1 complex, demonstrating different binding properties for different MHC molecules. RM 001, RM 003, RM 005 and RM 010 showed minimal or undetectable binding for a peptide/MHC complex without a peptide conjugate (p?/Al 1).
EXAMPLE 2
This Example provides a description of the binding properties of binding partners that bind with specificity to peptide conjugate/MHC complexes comprising MRTX849 or AMG- 510.
Binding kinetics of Fabs were assessed using an Octet RED96e instrument (Sartorious). Bio-layer interferometry (BLI) sensorgrams measured the interaction between the Fab and the peptide conjugate MHC complex. Biotinylated Fab was immobilized on streptavidin-coated BLI sensors to a final immobilization level ranging from 0.5 to 0.8 nm, and binding kinetics were measured against peptide conjugate/MHC complexes at 1, 4, 16, and 64 nmol/L in TBS buffer (FIG. 2A). The TBS buffer was pH 7.4, containing 1% BSA, 5 pmol/L biotin, and 0.005% Tween-20. In each BLI graph, the association step was from 0- 500 s and the dissociation step was from 500-1300 s.
An exemplary binding partner, RM 010, showed strong binding properties to peptide conjugate/MHC complex with either AMG-510 (sotorasib) or MRTX849 (adagrasib). RM 010 Fab bound to ada-p?/A03 complex with a dissociation constant (KD) of 4.8 nM. RM 010 Fab bound to ada-p?/Al 1 complex with a dissociation constant (KD) of 2.6 nM. RM 010 Fab bound to soto-p?/A03 complex with a dissociation constant (KD) of 21.3 nM. RM 010 Fab bound to soto-p?/Al 1 complex with a dissociation constant (KD) of 3.2 nM. The RM 010 Fab showed undetectable binding to the peptide/MHC complex in the absence of a targeted covalent inhibitor.
The exemplary binding partner, RM 010, showed minimal inhibition by free drug (e.g., free adagrasib) of the interaction between the Fab and ada-p?/Al 1 complex. Biotinylated Fab was immobilized on streptavidin-coated BLI sensors, and binding of soluble ada-p?/Al 1 in absence (0 pmol/L) or presence (0.1 pmol/L, 1 pmol/L, or 10 pmol/L) of free adagrasib was measured (FIG. 2B). BLI sensorgrams showed minimal inhibition by free adagrasib even at the largest concentration of 10 pmol/L. The association step of the binding signal was measured from 0-500 s followed by the dissociation step. A binding signal intensity was taken at 1200 s in the measurement. The signal was normalized using the value without free adagrasib (100%) and the 0 nM baseline (0%) to obtain a plot in function of free-adagrasib concentration (FIG. 2C).
EXAMPLE 3
This Example provides a description of how conjugations within p? affected binding of the R023 binder to peptide-conjugated sotorasib.
A single cysteine (e.g., Cys) residue was systematically introduced on all wild-type p? positions to replace the original residues, excluding the anchor positions necessary for MHC binding (FIG. 3A). For p? peptide, these were residues 8 and 16. The peptides were conjugated with sotorasib and then used to refold soto-p/MHC complexes with HLA-A*03 and HLA-A*11. Binder R023 bound MHCs presenting sotorasib from residues 10 to 12 of p?, but did not bind as efficiently when sotorasib was presented at residues 7, 13, 14, or 15 (FIGs. 3B-3C). R023 efficiently bound sotorasib presented at residue 9 in HLA-A*03 (top row of FIG. 3C) but not in HLA-A* 11 (bottom row of FIG. 3C).
EXAMPLE 4
This Example provides a description of how binder R023 could bind to modified soto- p? presented by other HLA molecules aside from HLA-A*03 and HLA-A* 11. Other tested HLA were HLA-A*01:01, HLA-A*02:01, HLA-A*24:02, and HLA-A*26:01.
The anchoring residues of the p? peptide sequence were modified to enable high- affinity binding to each HLA (FIG. 4A). Peptides were conjugated with sotorasib and refolded correctly paired soto-p/MHC complexes. R023 bound sotorasib-peptide conjugates presented by HLA-A*02:01, but not by the other tested HLAs (FIGs. 4B-4C). As shown in the BLI sensorgram, the Bmax was similar between the binding for HLA-A*02 and HLA- A*03. These results indicated that a modified p? peptide conjugated to sotorasib may be recognized by the R023 binder when complexed to a HLA that was not HLA-A*03 and HLA-A*11.
EXAMPLE 5
This Example provides a description of a mutational analysis of the RM 010 antibody.
A deep mutational scanning analysis of the RM 010 antibody was performed. Select residues of the VL and VH domains of RM 010 were diversified, one residue at a time, to construct a library in a yeast display format. The specific residues modified for VL are shown in FIG. 5A. The specific residues modified for VH are shown in FIG. 6A. The library was sorted for binding and a lack of binding for adagrasib-p7 in complex with HLA-A*03 (abbreviated as ada-p7/A03) was evaluated for mutated VL (FIG. 5B) or mutated VH (FIG. 6B). The library was also sorted for binding and a lack of binding for adagrasib-p7 in complex with HLA-A*11 (abbreviated as ada-p7/Al 1) was evaluated for mutated VL (FIG. 5C) and mutated VH (FIG. 6C). Enrichment of variants are displayed as heatmaps.
Permissible single-point mutations for VL and VH were characterized for RM 010 through the deep mutational scanning. Consensus sequence were determined for amino acid residue positions. A summary of the single-point mutations is found in Table E.
EXAMPLE 6
This Example provides a description of (i) how different targeted covalent inhibitors in peptide-conjugate/MHC complex affected binding of RM 010 binder and (ii) how conjugations within p? affected binding of the RM 010 binder to peptide-conjugated sotorasib.
Drug cross-reactivity was examined with peptide-conjugate/MHC complexes with either adagrasib (ada-p7/Al l), sotorasib (soto-p7/Al l), or divarasib (GDC6036-p7/Al l) covalently bound to the peptide. RM 010 showed different affinity to each peptide- conjugate/MHC complex, with the greatest binding response to ada-p7/Al 1 (FIG. 7).
For the sotorasib-scan, a single cysteine (e.g., Cys) residue was systematically introduced to all wild-type p? positions to replace the original residues, excluding the anchor positions (residues 8 and 16) necessary for MHC binding. The peptides were conjugated with sotorasib and then used to refold soto-p/MHC complexes with HLA-A*11. Binder RM 010 bound MHCs presenting sotorasib from residues 12, 9, and 15 to 12 of p?, but did not bind as efficiently when sotorasib was presented at residues 7, 14, 13, 10, or 11 (FIG. 8). Binder RM 010 showed the greatest binding response to MHC presenting sotorasib from residue 12.
EXAMPLE 7
This Example provides a description of dissociation tests to examine binding of antibody clones to peptide conjugate/MHC complexes comprising adagrasib-KRAS(G12C) peptide presented on HLA-A11 or HLA-A03.
From a second-generation yeast-display library constructed from the RM010 clone, new clones were identified. Yeast cells displaying an antibody in the scFv format were suspended in 1 mL BSS (1 mg/ml BSA + TBS) at 4xl04 cell/pL in 5 pL BSS, and were labelled by mixing with 5 pL mouse anti-V5 antibody (1/250 in BSS). The labelled yeast cells were mixed with 10 pL of 20 nM biotinylated target (ada-p7/HLA-Al 1 or ada-p7/HLA- A03) in BSS, resulting in the yeast cell density of IxlO4 cell/pL and 10 nM biotinylated target. The mixture was incubated at room temperature for 30 minutes. Then, cells were transferred to a 96 well filter plate and washed twice with 150 pL BSST (BSS + 0.1% Tween) by applying vacuum. For the dissociation experiment, 20 pL of 500 nM nonbiotinylated competitor (non-biotinylated ada-p7/Al 1) was added to each well and the plate was incubated at 30°C for 60 minutes. The cells were washed twice with 150 pL BSST. Then, 20 pL of secondary antibody mixture (1/100 anti-mouse FITC + 1/100 NAV-Dylight 650 in BSS) was added to each well and the plate was incubated at 4°C for 30 minutes. The cells were washed twice with 150 pL BSST and then resuspended in 100 pL BSS for each well and analyzed using an iQue flow cytometer (Sartorius).
Following 60 minute incubation, binders RM101, RM102, RM103, RM104, RM105, RM106, RM107, RM108, RM109, RM110, RM111, RM112, RM113, RM114, RM115, RM1 16, RM117, RM118 showed noticeably higher MFI compared to that from parent clone RM010, indicating improved binding (FIG. 9). Binding was examined for ada-p7/Al 1 and ada-p7/A03 across antibody clones. All antibody clones, excluding RM118, RM120, and RM121, showed higher signals for ada-p7/Al 1, indicating slower dissociation rates than that of RM010 (FIG. 10). Binder RM119 further showed higher MFI signal for binding to ada- p7/A03 than that of RM010.
Overall, antibody clones exhibited higher signals after 60 minutes of dissociation in the presence of a nonbiotinylated competitor (that prevents rebinding), indicating improved (slower) dissociation rates compared to parent clone RM010. While preferred embodiments of the present disclosure have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the disclosure. It should be understood that various alternatives to the embodiments of the present disclosure may be employed in practicing the present disclosure. It is intended that the following claims define the scope of the present disclosure and that methods and structures within the scope of these claims and their equi val ents b e covered thereby .

Claims

CLAIMS What is claimed is:
1. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain binds to (i) a first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 100 nM and (ii) a second peptide conjugate/MHC complex with a KD of at most about 100 nM, wherein the first peptide conjugate/MHC complex comprises:
(a) a first peptide conjugate, wherein the first peptide conjugate of the first peptide conjugate/MHC complex is a first peptide covalently linked to a first targeted covalent inhibitor or fragment thereof; and
(b) a first MHC; wherein the second peptide conjugate/MHC complex comprises:
(a) a second peptide conjugate, wherein the second peptide conjugate of the second peptide conjugate/MHC complex is a second peptide covalently linked to a second targeted covalent inhibitor or fragment thereof; and
(b) a second MHC; and wherein the first targeted covalent inhibitor is different from the second targeted covalent inhibitor.
2. The composition of claim 1, wherein the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 50 nM and/or (ii) the second peptide conjugate/MHC complex with a KD of at most about 50 nM.
3. The composition of claim 1, wherein the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 10 nM and/or (ii) the second peptide conjugate/MHC complex with a KD of at most about 10 nM.
4. The composition of claim 1, wherein the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 5 nM and/or (ii) the second peptide conjugate/MHC complex with a KD of at most about 5 nM.
5. The composition of any one of claims 1-4, wherein the first targeted covalent inhibitor is a covalent inhibitor of a RAS protein and the second targeted covalent inhibitor is a covalent inhibitor of a RAS protein.
6. The composition of any one of claims 1-5, wherein the first targeted covalent inhibitor is a covalent inhibitor of KRASG12C and the second targeted covalent inhibitor is a covalent inhibitor of KRASG12C.
7. The composition of any one of claims 1-6, wherein (i) the first targeted covalent inhibitor is adagrasib and the second targeted covalent inhibitor is sotorasib; (ii) the first targeted covalent inhibitor is adagrasib and the second targeted covalent inhibitor is divarasib; or (iii) the first targeted covalent inhibitor is divarasib and the second targeted covalent inhibitor is sotorasib.
8. The composition of any one of claims 1-7, wherein the antigen-binding domain
(i) does not bind to a free first targeted covalent inhibitor that is not conjugated to a peptide with a dissociation constant (KD) of less than 200 nM,
(ii) does not bind to the first peptide with a KD of less than 200 nM,
(iii) does not bind to a free first peptide conjugate that is not complexed with an MHC with a KD of less than 200 nM, or
(iv) any combination above.
9. The composition of claim 8, wherein the antigen-binding domain binds to the first peptide conjugate/MHC complex with a greater affinity than to the first peptide, the free first targeted covalent inhibitor, and/or the free first peptide conjugate.
10. The composition of claim 8 or 9, wherein the affinity of the antigen-binding domain for the first peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the antigen-binding domain for the first peptide or the free first targeted covalent inhibitor.
11. The composition of any one of claims 1-10, wherein the antigen-binding domain does not detectably bind to a complex of the first peptide with the first MHC, wherein the first peptide is not covalently bound to the first targeted covalent inhibitor or fragment thereof.
12. The composition of any one of claims 8-11, wherein the antigen-binding domain does not detectably bind to the free first targeted covalent inhibitor.
13. The composition of any one of claims 1-12, wherein the antigen-binding domain binds to a free first peptide conjugate with a dissociation constant KD that is at least 2.5 times more than a KD of the antibody or the antigen-binding fragment to the first peptide conjugate/MHC complex.
14. The composition of any one of claims 1-13, wherein the antigen-binding domain (i) does not bind to a free second targeted covalent inhibitor that is not conjugated to a peptide with a dissociation constant (KD) of less than 200 nM,
(ii) does not bind to the second peptide with a KD of less than 200 nM,
(iii) does not bind to a free second peptide conjugate that is not complexed with an MHC with a KD of less than 200 nM, or
(iv) any combination above.
15. The composition of claim 14, wherein the antigen-binding domain binds to the second peptide conjugate/MHC complex with a greater affinity than to the second peptide, the free second targeted covalent inhibitor, and/or the free second peptide conjugate.
16. The composition of claim 14 or 15, wherein the affinity of the antigen-binding domain for the second peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the antigen-binding domain for the second peptide or the free second targeted covalent inhibitor.
17. The composition of any one of claims 1-16, wherein the antigen-binding domain does not detectably bind to a complex of the second peptide with the second MHC, wherein the second peptide is not covalently bound to the second targeted covalent inhibitor or fragment thereof.
18. The composition of any one of claims 14-17, wherein the antigen-binding domain does not detectably bind to the free second targeted covalent inhibitor.
19. The composition of any one of claims 1-18, wherein the antigen-binding domain binds to a free second peptide conjugate with a dissociation constant (KD that is at least 2.5 times more than a KD of the antibody or the antigen-binding fragment to the second peptide conjugate/MHC complex.
20. The composition of any one of claims 1-19, wherein the first peptide and the second peptide comprise the same amino acid sequence.
21. The composition of any one of claims 1-20, wherein the first peptide and the second peptide consist of the same amino acid sequence.
22. The composition of claim 20 or 21, wherein the first peptide or the second peptide comprises the amino acid sequence of VVVGACGVGK.
23. The composition of any one of claims 1-22, wherein the first peptide and second peptide comprise a different amino acid sequence.
24. The composition of claim 23, wherein the first peptide and the second peptide comprise a shared contiguous amino acid sequence that is at least 3, 4, 5, 6, 7, 8, or 9 amino acids in length.
25. The composition of any one of claims 22-24, wherein the first peptide and/or the second peptide comprises an amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV.
26. The composition of any one of claims 1-25, wherein the first MHC or the second MHC is encoded by an HL A, wherein the HL A is HLA-A*02:01, HLA-A*03:01, or HLA-A* 11 :01.
27. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain binds to a peptide conjugate/MHC complex, wherein the peptide conjugate of the peptide conjugate/MHC complex is a peptide covalently linked to a targeted covalent inhibitor or fragment thereof and a MHC; and wherein the antigen-binding domain binds to the peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 50 nM; wherein the targeted covalent inhibitor is adagrasib.
28. The composition of claim 27, wherein the peptide conjugate/MHC complex is a first peptide conjugate/MHC complex comprising a first peptide conjugate comprising a first peptide and a first targeted covalent inhibitor or fragment thereof; and wherein the antigen-binding domain further binds to a second peptide conjugate/MHC complex comprising (a) a second peptide conjugate, wherein the second peptide conjugate of the second peptide conjugate/MHC complex is a second peptide covalently linked to a second targeted covalent inhibitor or fragment thereof; and (b) a second MHC.
29. The composition of claim 28, wherein the antigen-binding domain binds to the second peptide conjugate/MHC complex with a KD of at most about 50 nM.
30. The composition of claim 28, wherein the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 10 nM and (ii) the second peptide conjugate/MHC complex with a KD of at most about 10 nM.
31. The composition of claim 28, wherein the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 5 nM and (ii) the second peptide conjugate/MHC complex with a KD of at most about 5 nM.
32. The composition of any one of claims 28-31, wherein the second targeted covalent inhibitor is sotorasib or divarasib.
33. The composition of any one of claims 28-32, wherein the antigen-binding domain does not bind to a free first targeted covalent inhibitor with a dissociation constant (KD) of less than 200 nM.
34. The composition of claim 33, wherein the antigen-binding domain binds to the first peptide conjugate/MHC complex with a greater affinity than to the first peptide or the free first targeted covalent inhibitor.
35. The composition of claim 33 or 34, wherein the affinity of the antigen-binding domain for the first peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the antigen-binding domain for the first peptide or the free first targeted covalent inhibitor.
36. The composition of any one of claims 28-35, wherein the antigen-binding domain does not detectably bind to a complex of the first peptide with a first MHC, wherein the first peptide is not covalently bound to the first targeted covalent inhibitor or fragment thereof.
37. The composition of any one of claims 33-36, wherein the antigen-binding domain does not detectably bind to the free first targeted covalent inhibitor.
38. The composition of any one of claims 28-37, wherein the antigen-binding domain binds to a free first peptide conjugate with a dissociation constant KD that is at least 2.5 times more than a KD of the antibody or the antigen-binding fragment to the first peptide conjugate/MHC complex.
39. The composition of any one of claims 28-38, wherein the antigen-binding domain does not bind to a free second targeted covalent inhibitor with a dissociation constant (KD) of less than 200 nM.
40. The composition of claim 39, wherein the antigen-binding domain binds to the second peptide conjugate/MHC complex with a greater affinity than to the second peptide or the free second targeted covalent inhibitor.
41. The composition of claim 39 or 40, wherein the affinity of the antigen-binding domain for the second peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the antigen-binding domain for the second peptide or the free second targeted covalent inhibitor.
42. The composition of any one of claims 28-41, wherein the antigen-binding domain does not detectably bind to a complex of the second peptide with the second MHC, wherein the second peptide is not covalently bound to the second targeted covalent inhibitor or fragment thereof.
43. The composition of any one of claims 39-42, wherein the antigen-binding domain does not detectably bind to the free second targeted covalent inhibitor.
44. The composition of any one of claims 28-43, wherein the antigen-binding domain binds to a free second peptide conjugate with a dissociation constant ( o) that is at least 2.5 times more than a Ko of the antibody or the antigen-binding fragment to the second peptide conjugate/MHC complex.
45. The composition of any one of claims 28-44, wherein the first peptide and the second peptide comprise the same amino acid sequence.
46. The composition of any one of claims 28-45, wherein the first peptide and the second peptide consist of the same amino acid sequence.
47. The composition of claim 46, wherein the first peptide or the second peptide comprises the amino acid sequence of VVVGACGVGK.
48. The composition of any one of claims 28-44, wherein the first peptide and second peptide comprise a different amino acid sequence.
49. The composition of claim 48, wherein the first peptide or the second peptide comprises an amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV.
50. The composition of any one of claims 28-49, wherein the MHC or the second MHC is an HL A, and wherein the HL A i s HL A- A* 02 : 01 , HL A- A* 03 : 01 , or HL A- A* 11 :01.
51. The composition of any one of claims 1-26 and 28-50, wherein the first peptide or the second peptide comprises a nucleophilic or an electrophilic residue, wherein the residue comprises cysteine, aspartic acid, or arginine.
52. The composition of any one of claims 1-26 and 28-51, wherein the first peptide or the second peptide comprises a cysteine residue.
53. The composition of any one of claims 1-26 and 28-52, wherein the first peptide conjugate is formed by a covalent reaction between the first targeted covalent inhibitor and a cysteine residue in the first peptide.
54. The composition of any one of claims 1-26 and 28-53, wherein the second peptide conjugate is formed by a covalent reaction between the second targeted covalent inhibitor and a cysteine residue in the second peptide.
55. The composition of any one of claims 1-26 and 28-54, wherein the first peptide or the second peptide is a segment of a protein that is associated with a cancer, optionally wherein the protein is encoded by a gene that is mutated in a cancer.
56. The composition of any one of claims 1-26 and 28-55, wherein the first peptide or the second peptide is a segment of an enzyme, and wherein the first targeted covalent inhibitor or second targeted covalent inhibitor is an inhibitor of the enzyme.
57. The composition of claim 56, wherein the enzyme is a kinase or GTPase.
58. The composition of any one of claims 1-26 and 28-57, wherein the first peptide or the second peptide is or is derived from KRAS.
59. The composition of any one of claims 1-26 and 28-58, wherein the first peptide or the second peptide comprises a segment of KRASG12C, KRASG12D, KRASG12R, or KRASG12S.
60. The composition of any one of claims 1-26 and 28-59, wherein the first targeted covalent inhibitor or the second targeted covalent inhibitor is (i) a tri-complex KRASGI2C inhibitor or a KRASG12C degrader, (ii) a tri-complex KRASGI2D inhibitor or a KRASG12D degrader, (iii) a tri-complex KRASG12R inhibitor or a KRASG12R degrader, (iv) a tri-complex KRASG12S inhibitor or a KRASG12S degrader.
61. The composition of any one of claims 1-26 and 28-60, wherein the first peptide conjugate or the second peptide conjugate comprises a compound selected from the group consisting of compounds 1, 2, and 3:
wherein the compound is covalently bonded to a cysteine residue in a peptide, wherein the peptide comprises an amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV.
62. The composition of claim 61, wherein the peptide comprises the amino acid sequence of VVVGACGVGK or VVGACGVGK and the MHC is HLA-A*03:01 or HLA-
A*l l:01.
63. The composition of claim 61, wherein the peptide comprises the amino acid sequence of KLVVVGACGV and the MHC is HLA-A*02:01.
64. The composition of any one of claims 1-26 and 28-63, wherein the first peptide conjugate is formed by the covalent reaction of adagrasib with a KRASG12C peptide.
65. The composition of any one of claims 1-26 and 28-64, wherein the second peptide conjugate is formed by the covalent reaction of sotorasib or divarasib with a KRASG12C peptide
66. The composition of any one of claims 1-26 and 28-66, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR1 comprising an amino acid sequence of X1SX3X4SIH, wherein
Xi is I, F or V,
X3 is S or Y, and
XHs S or Y;
(ii) a HC CDR2 comprising an amino acid sequence of X1ISX4X5X6X7X8TX10YADSVKG, wherein Xi is S or Y, X4is S or P, Xs is S or Y, Xe is S or Y, X7 is S or G, Xs is S or Y, and Xio is S or Y, and
(iii) a HC CDR3 comprising an amino acid sequence of XNX1X2DY, wherein
XN IS an amino acid sequence selected from LWAS, FQWY, GYGW, GWYYL, YWYYM, YWYYL, GYYYPYY, SYYGFWQALW, SSRQYYHSQVEPPM, SGYYSSHWYLQSWYQ, and HYSEKWWGWYTMYID,
Xi is G or A, and
X2 is L or M; and/or
(b) the VL comprises:
(i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and
(iii) a LC CDR3 comprising the amino acid sequence of QQX3X4X5X6X7X8X9X10T, wherein
X3 is W, T, S, A or G,
X4is N, W, S, G, Y, D or K,
X5 is W, Y, S, A or T,
Xe is G, S, Y, L, W, E or D,
X7is W, S, H, Y, E, F or absent,
Xs is P, Q, S, W, E, L, G or absent, X9 is L or P, and
X10 is I, L, F, or V.
67. The composition of any one of claims 1-26 and 28-66, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of LWASGLDY; and/or
(b) the VL comprises:
(i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
68. The composition of any one of claims 1-26 and 28-66, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of SYYGFWQALWALDY; and/or
(b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
69. The composition of any one of claims 1-26 and 28-66, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of GYYYPYYAMDY; and/or
(b) the VL comprises:
(i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
70. The composition of any one of claims 1-26 and 28-66, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein
(a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of GYYYPYYAMDY; and/or
(b) the VL comprises:
(i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
71. The composition of any one of claims 1-26 and 28-66, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of X1X2X3X4X5MDY, wherein Xi is F or G, X2 is Q or Y, X3 is W or G, X4 is Y or W, and X5 is A or G; and/or
(b) the VL comprises:
(i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
72. The composition of any one of claims 1-26 and 28-66, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of SGYYSSHWYLQSWYQAMDY; and/or
(b) the VL comprises:
(i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
73. The composition of any one of claims 1-26 and 28-66, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of SGYYSSHWYLQSWYQAMDY; and/or
(b) the VL comprises:
(i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
74. The composition of any one of claims 1-26 and 28-66, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of HYSEKWWGWYTMYIDAMDY; and/or
(b) the VL comprises:
(i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
75. The composition of any one of claims 1-26 and 28-66, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of X1WYYX2GMDY, wherein Xi is G or Y and X2 is M or L; and/or
(b) the VL comprises:
(i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(iii) a LC CDR3 comprising the amino acid comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
76. The composition of any one of claims 1-26 and 28-66, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of X1WYYX2GMDY, wherein Xi is G or Y and X2 is M or L; and/or
(b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
77. The composition of any one of claims 1-26 and 28-66, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of X1WYYX2GMDY, wherein Xi is G or Y and X2 is M or L; and/or
(b) the VL comprises:
(i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(iii) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
78. The composition of any one of claims 1-26 and 28-66, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein
(a) the VH comprises: (i) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(ii) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(iii) a HC CDR3 comprising an amino acid sequence of SSRQYYHSQVEPPMAMDY; and/or
(b) the VL comprises:
(i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(iii) a LC CDR3 comprising the amino acid comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
79. The composition of any one of claims 1-26 and 28-78, wherein the antigen-binding domain comprises the HC CDR1, HC CDR2, and HC CDR3 amino acid sequences of a VH amino acid sequence and/or the LC CDR1, LC CDR2, and LC CDR3 amino acid sequences of a VL amino acid sequence of a polypeptide selected from the group consisting of RM_001, RM_002, RM_003, RM_004, RM_005, RM_006, RM_007, RM_008, RM_009, RM_010, RM_011, RM_012, RM_013, RM_014, RM_015, RM_016, RM_017, RM_018, RM_019, RM_020, RM_021, RM_022, , RM_023, RM_024, RM 025, RM_026, RM_027, RM_028 , RM_029, RM_030, RM_031, and RM 032; or a variant thereof comprising 1-5 amino acid changes in one or more of the CDR amino acid sequences.
80. The composition of any one of claims 1-26 and 28-79, wherein the antigen-binding domain comprises the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and/or LC CDR3 amino acid sequences of a polypeptide selected from the group consisting of RM_001, RM_002, RM_003, RM_004, RM_005, RM_006, RM_007, RM_008, RM_009, RM_010, RM_011, RM_012, RM_013, RM_014, RM_015, RM_016, RM_017, RM_018, RM_019, RM_020, RM_021, RM_022, , RM_023, RM_024, RM 025, RM_026, RM_027, RM_028 , RM_029, RM_030, RM_031, and RM 032, or a variant thereof comprising 1-5 amino acid changes in one or more of the CDR amino acid sequences.
81. The composition of any one of claims 1-26 and 28-80, wherein the antigen-binding domain comprises a VH and/or a VL amino acid sequence that is 90%, 95%, or 100% identical to the following VH and VL sequences:
RM_001
VL :DIQMTQ SP S SLS AS VGDRVTITCRASQ S VS S AV AW YQQKPGKAPKLLI YS AS SL YSGVPSRFSGSRSGTDFTL
TISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV
VH:EVQLVESGGGL VQPGGSLRLSC AASGFTIS S S SIHWVRQAPGKGLEWVASIS S Y YGSTSYADSVKGRFTISAD
TSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWGQGTLVTVSS
RM_002
VL :DIQMTQ SP S SLS AS VGDRVTITCRASQ S VS S AV AW YQQKPGKAPKLLI YS AS SL YSGVP
SRFSGSRSGTDFTL TISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV VH:EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS SY YGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGLDYWG QGTLVTVSS
RM_003
VL :DIQMTQ SP S SLS AS VGDRVTITCRASQ S VS S AV AW YQQKPGKAPKLLI YS AS SL YSGVPSRFSGSRSGTDFTL
TISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRTV
VH:EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S S SGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALW ALD YWGQGTL VT VS S
RM_004
VL :DIQMTQ SP S SLS AS VGDRVTITCRASQ S VS S AV AW YQQKPGKAPKLLI YS AS SL YSGVPSRFSGSRSGTDFTL
TISSLQPEDFATYYCQQWWYGSPLFTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISSS SGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQALW ALD YWGQGTL VT VS S
RM_005
VL :DIQMTQ SP S SLS AS VGDRVTITCRASQ S VS S AV AW YQQKPGKAPKLLI YS AS SL YSGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISS YYGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAM D YWGQGTL VTVSS
RM_006 VL :DIQMTQ SP S SLS AS VGDRVTITCRASQ S VS S AV AW YQQKPGKAPKLLI YS AS SL YSGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISP YYGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAM DYWGQGTLVTVSS
RM_007
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQTSWYHSLITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSY
YGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDY WGQGTLVTVSS
RM_008
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVASISSY YGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDY WGQGTLVTVSS
RM_009
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSY
YGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPYYAMDY WGQGTLVTVSS
RM_010
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSWLYWLVTFGQGTKVEIKRTV VH:EVQLVESGGGL VQPGGSLRLSC AASGFTF S S S SIHWVRQAPGKGLEWVASIS S SS GSTSYADSVKGRFTISAD
TSKNTAYLQMNSLRAEDTAVYYCARFQWYAMDYWGQGTLVTVSS
RM_011
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQWNWGWPLITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTISSYSIHWVRQAPGKGLEWVAYISSYS GYTSYADSVKGRFTISAD
TSKNTAYLQMNSLRAEDTAVYYCARGYGWGMDYWGQGTLVTVSS
RM_012
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTISYSSIHWVRQAPGKGLEWVAYISSSS GYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQS W YQ AMD YWGQGTL VT VS S RM_013
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQASYGPITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISSY
YGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSHWYLQS W YQ AMD YWGQGTL VT VS S
RM_014
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQWWSSSQLITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVASISSY
YGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARHYSEKWWGWY TMYID AMD YWGQGTL VT VS S
RM_015
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPY
SGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYW
GQGTLVTVSS
RM_016
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGKTYYPITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWG
QGTLVTVSS
RM_017
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPY
SGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYW
GQGTLVTVSS
RM_018
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPY
SGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYW GQGTLVTVSS
RM_019
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYWG QGTLVTVSS
RM_020
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYISPY
SGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLGMDYW
GQGTLVTVSS
RM_021
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG
QGTLVTVSS
RM_022
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG
QGTLVTVSS
RM_023
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSSYYEELITFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG
QGTLVTVSS
RM_024
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQAYSDPLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG
QGTLVTVSS
RM 025
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY
SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV
VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS
SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG
QGTLVTVSS
RM_026 VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSGSYLLITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG QGTLVTVSS
RM_027
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQADYEFGLITFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISPYS SYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMGMDYWG QGTLVTVSS
RM_028
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVAYISSSY GYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPP MAMD YWGQGTL VT VS S
RM_029
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVAYISSS YGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEP PMAMD YWGQGTL VT VS S
RM_030
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYISSSY GYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEPP MAMD YWGQGTL VT VS S
RM_031
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSC AASGFT VS YS SIHWVRQAPGKGLEWVAYIS S S YGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYYHSQVEP PMAMD YWGQGTL VT VS S
RM_032
VL:DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLY SGVPSRFSGSRSGTDFTL TISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKRTV VH:EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYISPY SGYTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYLGMDYW GQGTLVTVSS
82. The composition of any one of claims 1-26 and 28-81, wherein the polypeptide specifically binds to the first peptide conjugate/MHC complex and a T cell antigen.
83. The composition of claim 82, wherein the polypeptide specifically binds to the first peptide conjugate/MHC complex and human CD3.
84. The composition of any one of claims 1-26 and 28-83, wherein the polypeptide specifically binds to the second peptide conjugate/MHC complex and a T cell antigen.
85. The composition of claim 84, wherein the polypeptide specifically binds to the second peptide conjugate/MHC complex and human CD3.
86. The composition of any one of claims 1-85, wherein the polypeptide comprises a heavy chain constant region selected from the group consisting of human IgM, IgGi, IgG2, IgGs, IgG4, IgAi, and IgA.
87. The composition of claim 86, wherein the heavy chain constant region comprises one or more amino acid substitutions, deletions, or additions in the Fc region.
88. The composition of any one of claims 1-87, wherein the polypeptide is conjugated to a detectable label, a chemotherapeutic agent, a radioisotope, an enzyme, or a toxin.
89. The composition of any one of claims 1-88, wherein the polypeptide is comprised within a chimeric antigen receptor.
90. The composition of claim 89, wherein the polypeptide is expressed by a T cell, a killer macrophage, a neutrophil or a natural killer cell.
91. A polynucleotide encoding the polypeptide of any one of claims 1-90.
92. A polynucleotide encoding a heavy chain variable region and/or a light chain variable region of the polypeptide of any one of claims 1-91.
93. A vector comprising the polynucleotide of claim 91 or 92.
94. The vector of claim 93, wherein the vector is a viral vector.
95. The vector of claim 94, wherein the viral vector is an adenoviral vector, lentiviral vector, retroviral vector, or adeno-associated viral vector.
96. A recombinant host cell comprising:
(a) the polynucleotide of claim 91 or 92;
(b) the vector of any one of claims 93-95;
(c) a first polynucleotide encoding a VH or a heavy chain of the polypeptide of any one of claims 1-90, and a second polynucleotide encoding a VL or a light chain of the polypeptide of any one of claims 1-90; or
(d) a first vector comprising a first polynucleotide encoding a VH or a heavy chain of the polypeptide of any one of claims 1-90, and a second vector comprising a second polynucleotide encoding a VL or a light chain of the polypeptide of any one of claims 1-90.
97. A pharmaceutical composition comprising the polypeptide of any one of claims 1-90, the polynucleotide of claim 91 or 92, the vector of any one of claims 93-95, or the host cell of claim 96 and a pharmaceutically acceptable carrier or excipient.
98. A method of producing a polypeptide, the method comprising culturing the host cell of claim 96 under suitable conditions so that the polynucleotide is expressed and the binding partner is produced.
99. An eukaryotic cell comprising the polynucleotide of claim 91 or 92 or the vector of any one of claims 93-95, wherein the cell is optionally a totipotent, multipotent, or pluripotent stem cell, wherein optionally the stem cell has an induced stem cell phenotype, or wherein the cell is optionally a leukocyte, optionally a CD4+ T cell, optionally a CD8+ T cell, optionally a y5 T cell, optionally a natural killer cell , a neutrophil or a macrophage.
100. A method comprising administering to an individual in need thereof the polypeptide of any one of claims 1-90, the polynucleotide of claim 91 or 92, the vector of any one of claims 93-95, the pharmaceutical composition of claim 97, or the cell of claim 96.
101. A cell free peptide conjugate/MHC complex comprising:
(a) an isolated peptide conjugate formed by the covalent reaction of a targeted covalent inhibitor with a peptide; and
(b) an MHC, wherein the targeted covalent inhibitor is sotorasib, adagrasib, or divarasib, wherein the MHC is an HL A, and wherein the HLA is HLA-A*02:01, HLA-A*03:01, or HLA-A*11:01.
102. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR3 comprising an amino acid sequence of XNX1X2DY, wherein
X\ is an amino acid sequence selected from LWAS, FQWY, GYGW, GWYYL, YWYYM, YWYYL, GYYYPYY, SYYGFWQALW, SSRQYYHSQVEPPM, SGYYSSHWYLQSWYQ, and
HYSEKWWGWYTMYID,
Xi is G or A, and
X2 is L or M; and/or
(b) the VL comprises:
(i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and
(iii) a LC CDR3 comprising the amino acid sequence of QQX3X4X5X6X7X8X9X10T, wherein
X3 is W, T, S, A or G,
X4 is N, W, S, G, Y, D or K,
X5 is W, Y, S, A or T,
Xe is G, S, Y, L, W, E or D,
X7 is W, S, H, Y, E, F or absent,
Xs is P, Q, S, W, E, L, G or absent,
X9 is L or P, and
X10 is I, L, F, or V.
103. The composition of claim 102, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of X1SX3X4SIH, wherein
Xi is I, F or V,
X3 is S or Y, and
X4 is S or Y.
104. The composition of claim 102 or 103, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of X1ISX4X5X6X7X8TX10YADSVKG, wherein
Xi is S or Y,
X4 is S or P,
X5 is S or Y,
Xe is S or Y,
X7 is S or G,
Xs is S or Y, and
Xio is S or Y.
105. The composition of any one of claims 102-104, wherein the VL comprises a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
106. The composition of any one of claims 102-105, wherein the VL comprises a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
107. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR1 comprising an amino acid sequence of X1SX3X4SIH, wherein
Xi is I, F or V,
X3 is S or Y, and
XHs S or Y;
(ii) a HC CDR2 comprising an amino acid sequence of X1ISX4X5X6X7X8TX10YADSVKG, wherein
Xi is S or Y,
X4is S or P,
X5 is S or Y,
Xe is S or Y,
X?is S or G,
Xs is S or Y, and
Xio is S or Y, and
(iii) a HC CDR3 comprising an amino acid sequence of XNX1X2DY, wherein
XN IS an amino acid sequence selected from LWAS, FQWY, GYGW, GWYYL, YWYYM, YWYYL, GYYYPYY, SYYGFWQALW, SSRQYYHSQVEPPM, SGYYSSHWYLQSWYQ, and
HYSEKWWGWYTMYID,
Xi is G or A, and
X2 is L or M; and/or
(b) the VL comprises: (i) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(ii) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and
(iii) a LC CDR3 comprising the amino acid sequence of QQX3X4X5X6X7X8X9X10T, wherein
X3 is W, T, S, A or G, X4is N, W, S, G, Y, D or K, X5 is W, Y, S, A or T, Xe is G, S, Y, L, W, E or D, X7is W, S, H, Y, E, F or absent, Xs is P, Q, S, W, E, L, G or absent, X9 is L or P, and
X10 is I, L, F, or V.
108. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of LWASGLDY (SEQ ID NO: 5).
109. The composition of any one of claims 102-108, wherein the VH comprises a HC CDR2 sequence of SISSYYGSTSYADSVKG (SEQ ID NO: 4).
110. The composition of any one of claims 102-109, wherein the VH comprises a HC CDR1 sequence of ISSSSIH (SEQ ID NO: 3).
111. The composition of any one of claims 102-110, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQWNWGWPLIT (SEQ ID NO: 8).
112. The composition of any one of claims 102-111, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 7).
113. The composition of any one of claims 102-112, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 6).
114. The composition of any one of claims 102-113, wherein the antigen-binding domain comprises: a HC CDR1 sequence of ISSSSIH (SEQ ID NO: 3), a HC CDR2 sequence of SISSYYGSTSYADSVKG (SEQ ID NO: 4), a HC CDR3 sequence of LWASGLDY (SEQ ID NO: 5), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 6), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 7), and a LC CDR3 sequence of QQWNWGWPLIT (SEQ ID NO: 8).
115. The composition of any one of claims 102-114, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTIS S S SIHWVRQAPGKGLEWVASIS S YYGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGL DYWGQGTLVTVSS (SEQ ID NO: 2).
116. The composition of any one of claims 102-115, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKV EIKRTV (SEQ ID NO: 1).
117. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of LWASGLDY (SEQ ID NO: 13).
118. The composition of claim 117, wherein the VH comprises a HC CDR2 sequence of SISSYYGSTSYADSVKG (SEQ ID NO: 12).
119. The composition of claim 117 or 118, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 11).
120. The composition of any one of claims 117-119, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQWNWGWPLIT (SEQ ID NO: 16).
121. The composition of any one of claims 117-120, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 15).
122. The composition of any one of claims 117-121, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 14).
123. The composition of any one of claims 117-122, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 11), a HC CDR2 sequence of SISSYYGSTSYADSVKG (SEQ ID NO: 12), a HC CDR3 sequence of LWASGLDY (SEQ ID NO: 13), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 14), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 15), and a LC CDR3 sequence of QQWNWGWPLIT (SEQ ID NO: 16).
124. The composition of any one of claims 117-123, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S YYGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARLWASGL DYWGQGTLVTVSS (SEQ ID NO: 10).
125. The composition of any one of claims 117-124, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKV EIKRTV (SEQ ID NO: 9).
126. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SYYGFWQALWALDY (SEQ ID NO: 21).
127. The composition of claim 126, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 20).
128. The composition of claim 126 or 127, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 19).
129. The composition of any one of claims 126-128, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQWWYGSPLFT (SEQ ID NO: 24).
130. The composition of any one of claims 126-129, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 23).
131. The composition of any one of claims 126-130, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 22).
132. The composition of any one of claims 126-131, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 19), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 20), a HC CDR3 sequence of SYYGFWQALWALDY (SEQ ID NO: 21), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 22), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 23), and a LC CDR3 sequence of QQWWYGSPLFT (SEQ ID NO: 24).
133. The composition of any one of claims 126-132, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQ ALWALDYWGQGTLVTVSS (SEQ ID NO: 18).
134. The composition of any one of claims 126-133, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWYGSPLFTFGQGTKV EIKRT (SEQ ID NO: 17).
135. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SYYGFWQALWALDY (SEQ ID NO: 29).
136. The composition of claim 135, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 28).
137. The composition of claim 135 or 136, wherein the VH comprises a HC CDR1 sequence of FSSYSIH (SEQ ID NO: 27).
138. The composition of any one of claims 135-137, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQWWYGSPLFT (SEQ ID NO: 32).
139. The composition of any one of claims 135-138, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 31).
140. The composition of any one of claims 135-139, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 30).
141. The composition of any one of claims 135-140, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSYSIH (SEQ ID NO: 27), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 28), a HC CDR3 sequence of SYYGFWQALWALDY (SEQ ID NO: 29), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 30), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 31), and a LC CDR3 sequence of QQWWYGSPLFT (SEQ ID NO: 32).
142. The composition of any one of claims 135-141, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S YSIHWVRQAPGKGLEW VASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSYYGFWQ ALWALDYWGQGTLVTVSS (SEQ ID NO: 26).
143. The composition of any one of claims 135-142, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWYGSPLFTFGQGTKV EIKRT (SEQ ID NO: 25).
144. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GYYYP YYAMD Y (SEQ ID NO: 37).
145. The composition of claim 144, wherein the VH comprises a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 36).
146. The composition of claim 144 or 145, wherein the VH comprises a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 35).
147. The composition of any one of claims 144-146, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGKTYYPIT (SEQ ID NO: 40).
148. The composition of any one of claims 144-147, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 39).
149. The composition of any one of claims 144-148, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 38).
150. The composition of any one of claims 144-149, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 35), a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 36), a HC CDR3 sequence of GYYYP YYAMD Y (SEQ ID NO: 37), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 38), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 39), and a LC CDR3 sequence of QQGKTYYPIT (SEQ ID NO: 40).
151. The composition of any one of claims 144-150, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPY YAMDYWGQGTLVTVSS (SEQ ID NO: 34).
152. The composition of any one of claims 144-151, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEI KRTV (SEQ ID NO: 33).
153. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GYYYP YYAMD Y (SEQ ID NO: 45).
154. The composition of claim 153, wherein the VH comprises a HC CDR2 sequence of SISPYYGSTYYADSVKG (SEQ ID NO: 44).
155. The composition of claim 153 or 154, wherein the VH comprises a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 43).
156. The composition of any one of claims 153-155, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGKTYYPIT (SEQ ID NO: 48).
157. The composition of any one of claims 153-156, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 47).
158. The composition of any one of claims 153-157, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 46).
159. The composition of any one of claims 153-158, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 43), a HC CDR2 sequence of SISPYYGSTYYADSVKG (SEQ ID NO: 44), a HC CDR3 sequence of GYYYP YYAMD Y (SEQ ID NO: 45), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 46), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 47), and a LC CDR3 sequence of QQGKTYYPIT (SEQ ID NO: 48).
160. The composition of any one of claims 153-159, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVASISP YYGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPY YAMDYWGQGTLVTVSS (SEQ ID NO: 42).
161. The composition of any one of claims 153-160, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEI KRTV (SEQ ID NO: 41).
162. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GYYYP YYAMD Y (SEQ ID NO: 53).
163. The composition of claim 162, wherein the VH comprises a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 52).
164. The composition of claim 162 or 163, wherein the VH comprises a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 51).
165. The composition of any one of claims 162-164, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQTSWYHSLIT (SEQ ID NO: 56).
166. The composition of any one of claims 162-165, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 55).
167. The composition of any one of claims 162-166, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 54).
168. The composition of any one of claims 162-167, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 51), a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 52), a HC CDR3 sequence of GYYYP YYAMD Y (SEQ ID NO: 53), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 54), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 55), and a LC CDR3 sequence of QQTSWYHSLIT (SEQ ID NO: 56).
169. The composition of any one of claims 162-168, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPY YAMDYWGQGTLVTVSS (SEQ ID NO: 50).
170. The composition of any one of claims 162-169, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQTSWYHSLITFGQGTKVE IKRTV (SEQ ID NO: 49).
171. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GYYYP YYAMD Y (SEQ ID NO: 61).
172. The composition of claim 171, wherein the VH comprises a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 60).
173. The composition of claim 171 or 172, wherein the VH comprises a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 59).
174. The composition of any one of claims 171-173, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGKTYYPIT (SEQ ID NO: 64).
175. The composition of any one of claims 171-174, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 63).
176. The composition of any one of claims 171-175, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 62).
177. The composition of any one of claims 171-176, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 59), a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 60), a HC CDR3 sequence of GYYYP YYAMD Y (SEQ ID NO: 61), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 62), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 63), and a LC CDR3 sequence of QQGKTYYPIT (SEQ ID NO: 64).
178. The composition of any one of claims 171-177, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVASIS SYYGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYP YYAMD YWGQGTLVTVSS (SEQ ID NO: 58).
179. The composition of any one of claims 171-178, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEI KRTV (SEQ ID NO: 57).
180. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GYYYP YYAMD Y (SEQ ID NO: 69).
181. The composition of claim 180, wherein the VH comprises a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 68).
182. The composition of claim 180 or 181, wherein the VH comprises a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 67).
183. The composition of any one of claims 180-182, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 72).
184. The composition of any one of claims 180-183, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 71).
185. The composition of any one of claims 180-184, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 70).
186. The composition of any one of claims 180-185, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 67), a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 68), a HC CDR3 sequence of GYYYP YYAMD Y (SEQ ID NO: 69), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 70), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 71), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 72).
187. The composition of any one of claims 180-186, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYYYPY YAMDYWGQGTLVTVSS (SEQ ID NO: 66).
188. The composition of any one of claims 180-187, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEI KRTV (SEQ ID NO: 65).
189. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of FQWYAMDY (SEQ ID NO: 77).
190. The composition of claim 189, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 76).
191. The composition of claim 189 or 190, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 75).
192. The composition of any one of claims 189-191, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWLVT (SEQ ID NO: 80).
193. The composition of any one of claims 189-192, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 79).
194. The composition of any one of claims 189-193, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 78).
195. The composition of any one of claims 189-194, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 75), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 76), a HC CDR3 sequence of FQWYAMDY (SEQ ID NO: 77), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 78), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 79), and a LC CDR3 sequence of QQSSWLYWLVT (SEQ ID NO: 80).
196. The composition of any one of claims 189-195, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFQWYAM DYWGQGTLVTVSS (SEQ ID NO: 74).
197. The composition of any one of claims 189-196, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWLVTFGQGTKV EIKRTV (SEQ ID NO: 73).
198. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GYGWGMDY (SEQ ID NO: 85).
199. The composition of claim 198, wherein the VH comprises a HC CDR2 sequence of YISSYSGYTSYADSVKG (SEQ ID NO: 84).
200. The composition of claim 198 or 199, wherein the VH comprises a HC CDR1 sequence of ISSYSIH (SEQ ID NO: 83).
201. The composition of any one of claims 198-200, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQWNWGWPLIT (SEQ ID NO: 88).
202. The composition of any one of claims 198-201, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 87).
203. The composition of any one of claims 198-202, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 86).
204. The composition of any one of claims 198-203, wherein the antigen-binding domain comprises: a HC CDR1 sequence of ISSYSIH (SEQ ID NO: 83), a HC CDR2 sequence of YISSYSGYTSYADSVKG (SEQ ID NO: 84), a HC CDR3 sequence of GYGWGMDY (SEQ ID NO: 85), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 86), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 87), and a LC CDR3 sequence of QQWNWGWPLIT (SEQ ID NO: 88).
205. The composition of any one of claims 198-204, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTIS S YSMWVRQAPGKGLEWVAYIS S YSGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYGWG MDYWGQGTLVTVSS (SEQ ID NO: 82).
206. The composition of any one of claims 198-205, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWNWGWPLITFGQGTKV EIKRTV (SEQ ID NO: 81).
207. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SGYYSSHWYLQSWYQAMDY (SEQ ID NO: 93).
208. The composition of claim 207, wherein the VH comprises a HC CDR2 sequence of YISSSSGYTSYADSVKG (SEQ ID NO: 92).
209. The composition of claim 207 or 208, wherein the VH comprises a HC CDR1 sequence of ISYS SIH (SEQ ID NO: 91).
210. The composition of any one of claims 207-209, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSAWYPVT (SEQ ID NO: 96).
211. The composition of any one of claims 207-210, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 95).
212. The composition of any one of claims 207-211, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 94).
213. The composition of any one of claims 207-212, wherein the antigen-binding domain comprises: a HC CDR1 sequence of ISYS SIH (SEQ ID NO: 91), a HC CDR2 sequence of YISSSSGYTSYADSVKG (SEQ ID NO: 92), a HC CDR3 sequence of SGYYSSHWYLQSWYQAMDY (SEQ ID NO: 93), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 94), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 95), and a LC CDR3 sequence of QQSSAWYPVT (SEQ ID NO: 96).
214. The composition of any one of claims 207-213, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTIS YS SMWVRQAPGKGLEWVAYIS S SSGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSSH WYLQSWYQAMDYWGQGTLVTVSS (SEQ ID NO: 90).
215. The composition of any one of claims 207-214, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEI KRTV (SEQ ID NO: 89).
216. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SGYYSSHWYLQSWYQAMDY (SEQ ID NO: 101).
217. The composition of claim 216, wherein the VH comprises a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 100).
218. The composition of claim 216 or 217, wherein the VH comprises a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 99).
219. The composition of any one of claims 216-218, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASYGPIT (SEQ ID NO: 104).
220. The composition of any one of claims 216-219, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 103).
221. The composition of any one of claims 216-220, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 102).
222. The composition of any one of claims 216-221, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 99), a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 100), a HC CDR3 sequence of SGYYSSHWYLQSWYQAMDY (SEQ ID NO: 101), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 102), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 103), and a LC CDR3 sequence of QQASYGPIT (SEQ ID NO: 104).
223. The composition of any one of claims 216-222, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S YYSIHWVRQAPGKGLEWVASIS S YYGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSGYYSS HWYLQSWYQAMDYWGQGTLVTVSS (SEQ ID NO: 98).
224. The composition of any one of claims 216-223, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYGPITFGQGTKVEIKR TV (SEQ ID NO: 97).
225. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of HYSEKWWGWYTMYIDAMDY (SEQ ID NO: 109).
226. The composition of claim 225, wherein the VH comprises a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 108).
227. The composition of claim 225 or 226, wherein the VH comprises a HC CDR1 sequence of VSYSSIH (SEQ ID NO: 107).
228. The composition of any one of claims 225-227, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQWWSSSQLIT (SEQ ID NO: 112).
229. The composition of any one of claims 225-228, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 111).
230. The composition of any one of claims 225-229, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 110).
231. The composition of any one of claims 225-230, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSYSSIH (SEQ ID NO: 107), a HC CDR2 sequence of SISSYYGSTYYADSVKG (SEQ ID NO: 108), a HC CDR3 sequence of HYSEKWWGWYTMYIDAMDY (SEQ ID NO: 109), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 110), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 111), and a LC CDR3 sequence of QQWWSSSQLIT (SEQ ID NO: 112).
232. The composition of any one of claims 225-231, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVASISS YYGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARHYSEKW WGWYTMYIDAMDYWGQGTLVTVSS (SEQ ID NO: 106).
233. The composition of any one of claims 225-232, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQWWSSSQLITFGQGTKVE IKRTV (SEQ ID NO: 105).
234. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GWYYLGMDY (SEQ ID NO: 117).
235. The composition of claim 234, wherein the VH comprises a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 116).
236. The composition of claim 234 or 235, wherein the VH comprises a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 115).
237. The composition of any one of claims 234-236, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGKTYYPIT (SEQ ID NO: 120).
238. The composition of any one of claims 234-237, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 119).
239. The composition of any one of claims 234-238, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 118).
240. The composition of any one of claims 234-239, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 115), a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 116), a HC CDR3 sequence of GWYYLGMDY (SEQ ID NO: 117), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 118), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 119), and a LC CDR3 sequence of QQGKTYYPIT (SEQ ID NO: 120).
241. The composition of any one of claims 234-240, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYIS PYSGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYL GMDYWGQGTLVTVSS (SEQ ID NO: 114).
242. The composition of any one of claims 234-241, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEI KRTV (SEQ ID NO: 113).
243. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GWYYLGMDY (SEQ ID NO: 125).
244. The composition of claim 243, wherein the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 124).
245. The composition of claim 243 or 244, wherein the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 123).
246. The composition of any one of claims 243-245, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGKTYYPIT (SEQ ID NO: 128).
247. The composition of any one of claims 243-246, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 127).
248. The composition of any one of claims 243-247, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 126).
249. The composition of any one of claims 243-248, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 123), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 124), a HC CDR3 sequence of GWYYLGMDY (SEQ ID NO: 125), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 126), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 127), and a LC CDR3 sequence of QQGKTYYPIT (SEQ ID NO: 128).
250. The composition of any one of claims 243-249, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISP YSSYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLG MDYWGQGTLVTVSS (SEQ ID NO: 122).
251. The composition of any one of claims 243-250, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGKTYYPITFGQGTKVEI KRTV (SEQ ID NO: 121).
252. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GWYYLGMDY (SEQ ID NO: 133).
253. The composition of claim 252, wherein the VH comprises a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 132).
254. The composition of claim 252 or 253, wherein the VH comprises a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 131).
255. The composition of any one of claims 252-254, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 136).
256. The composition of any one of claims 252-255, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 135).
257. The composition of any one of claims 252-256, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 134).
258. The composition of any one of claims 252-257, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 131), a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 132), a HC CDR3 sequence of GWYYLGMDY (SEQ ID NO: 133), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 134), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 135), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 136).
259. The composition of any one of claims 252-258, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYIS PYSGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYL GMDYWGQGTLVTVSS (SEQ ID NO: 130).
260. The composition of any one of claims 252-259, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEI KRTV (SEQ ID NO: 129).
261. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GWYYLGMDY (SEQ ID NO: 141).
262. The composition of claim 261, wherein the VH comprises a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 140).
263. The composition of claim 261 or 262, wherein the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 139).
264. The composition of any one of claims 261-263, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 144).
265. The composition of any one of claims 261-264, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 143).
266. The composition of any one of claims 261-265, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 142).
267. The composition of any one of claims 261-266, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 139), a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 140), a HC CDR3 sequence of GWYYLGMDY (SEQ ID NO: 141), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 142), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 143), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 144).
268. The composition of any one of claims 261-267, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYIS PYSGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYL GMDYWGQGTLVTVSS (SEQ ID NO: 138).
269. The composition of any one of claims 261-268, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEI KRTV (SEQ ID NO: 137).
270. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GWYYLGMDY (SEQ ID NO: 149).
271. The composition of claim 270, wherein the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 148).
272. The composition of claim 270 or 271, wherein the VH comprises a HC CDR1 sequence of FSSYSIH (SEQ ID NO: 147).
273. The composition of any one of claims 270-272, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 152).
274. The composition of any one of claims 270-273, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 151).
275. The composition of any one of claims 270-274, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 150).
276. The composition of any one of claims 270-275, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSYSIH (SEQ ID NO: 147), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 148), a HC CDR3 sequence of GWYYLGMDY (SEQ ID NO: 149), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 150), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 151), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 152).
277. The composition of any one of claims 270-276, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTF S S YSIHWVRQAPGKGLEW VASISP YSSYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYLG MDYWGQGTLVTVSS (SEQ ID NO: 146).
278. The composition of any one of claims 270-277, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEI KRTV (SEQ ID NO: 145).
279. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of GWYYLGMDY (SEQ ID NO: 157).
280. The composition of claim 279, wherein the VH comprises a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 156).
281. The composition of claim 279 or 280, wherein the VH comprises a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 155).
282. The composition of any one of claims 279-281, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSAWYPVT (SEQ ID NO: 160).
283. The composition of any one of claims 279-282, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 159).
284. The composition of any one of claims 279-283, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 158).
285. The composition of any one of claims 279-284, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSYYSIH (SEQ ID NO: 155), a HC CDR2 sequence of YISPYSGYTSYADSVKG (SEQ ID NO: 156), a HC CDR3 sequence of GWYYLGMDY (SEQ ID NO: 157), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 158), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 159), and a LC CDR3 sequence of QQSSAWYPVT (SEQ ID NO: 160).
286. The composition of any one of claims 279-285, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYYSIHWVRQAPGKGLEWVAYIS PYSGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGWYYL GMDYWGQGTLVTVSS (SEQ ID NO: 154).
287. The composition of any one of claims 279-286, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEI KRTV (SEQ ID NO: 153).
288. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 165).
289. The composition of claim 288, wherein the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 164).
290. The composition of claim 288 or 289, wherein the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 163).
291. The composition of any one of claims 288-290, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 168).
292. The composition of any one of claims 288-291, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 167).
293. The composition of any one of claims 288-292, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 166).
294. The composition of any one of claims 288-293, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 163), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 164), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 165), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 166), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 167), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 168).
295. The composition of any one of claims 288-294, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISP YSSYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMG MDYWGQGTLVTVSS (SEQ ID NO: 162).
296. The composition of any one of claims 288-295, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEI KRTV (SEQ ID NO: 161).
297. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 173).
298. The composition of claim 297, wherein the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 172).
299. The composition of claim 297 or 298, wherein the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 171).
300. The composition of any one of claims 297-299, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSAWYPVT (SEQ ID NO: 176).
301. The composition of any one of claims 297-300, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 175).
302. The composition of any one of claims 297-301, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 174).
303. The composition of any one of claims 297-302, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 171), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 172), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 173), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 174), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 175), and a LC CDR3 sequence of QQSSAWYPVT (SEQ ID NO: 176).
304. The composition of any one of claims 297-303, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISP YSSYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMG MDYWGQGTLVTVSS (SEQ ID NO: 170).
305. The composition of any one of claims 297-304, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSAWYPVTFGQGTKVEI KRTV (SEQ ID NO: 169).
306. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 181).
307. The composition of claim 306, wherein the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 180).
308. The composition of claim 306 or 307, wherein the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 179).
309. The composition of any one of claims 306-308, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSYYEELIT (SEQ ID NO: 184).
310. The composition of any one of claims 306-309, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 183).
311. The composition of any one of claims 306-310, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 182).
312. The composition of any one of claims 306-311, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 179), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 180), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 181), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 182), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 183), and a LC CDR3 sequence of QQSSYYEELIT (SEQ ID NO: 184).
313. The composition of any one of claims 306-312, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISP YSSYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMG MDYWGQGTLVTVSS (SEQ ID NO: 178).
314. The composition of any one of claims 306-313, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSYYEELITFGQGTKVEI KRTV (SEQ ID NO: 177).
315. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 189).
316. The composition of claim 315, wherein the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 188).
317. The composition of claim 315 or 316, wherein the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 187).
318. The composition of any one of claims 315-317, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQAYSDPLT (SEQ ID NO: 192).
319. The composition of any one of claims 315-318, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 191).
320. The composition of any one of claims 315-319, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 190).
321. The composition of any one of claims 315-320, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 187), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 188), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 189), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 190), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 191), and a LC CDR3 sequence of QQAYSDPLT (SEQ ID NO: 192).
322. The composition of any one of claims 315-321, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISP YSSYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMG MDYWGQGTLVTVSS (SEQ ID NO: 186).
323. The composition of any one of claims 315-322, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQAYSDPLTFGQGTKVEIK RTV (SEQ ID NO: 185).
324. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 197).
325. The composition of claim 324, wherein the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 196).
326. The composition of claim 324 or 325, wherein the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 195).
327. The composition of any one of claims 324-326, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGSSSLLT (SEQ ID NO: 200).
328. The composition of any one of claims 324-327, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 199).
329. The composition of any one of claims 324-328, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 198).
330. The composition of any one of claims 324-329, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 195), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 196), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 197), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 198), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 199), and a LC CDR3 sequence of QQGSSSLLT (SEQ ID NO: 200).
331. The composition of any one of claims 324-330, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISP YSSYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMG MDYWGQGTLVTVSS (SEQ ID NO: 194).
332. The composition of any one of claims 324-331, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKR TV (SEQ ID NO: 193).
333. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 205).
334. The composition of claim 333, wherein the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 204).
335. The composition of claim 333 or 334, wherein the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 203).
336. The composition of any one of claims 333-335, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSGSYLLIT (SEQ ID NO: 208).
337. The composition of any one of claims 333-336, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 207).
338. The composition of any one of claims 333-337, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 206).
339. The composition of any one of claims 333-338, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 203), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 204), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 205), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 206), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 207), and a LC CDR3 sequence of QQSGSYLLIT (SEQ ID NO: 208).
340. The composition of any one of claims 333-339, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISP YSSYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMG MDYWGQGTLVTVSS (SEQ ID NO: 202).
341. The composition of any one of claims 333-340, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSGSYLLITFGQGTKVEIK RTV (SEQ ID NO: 201).
342. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYMGMDY (SEQ ID NO: 213).
343. The composition of claim 342, wherein the VH comprises a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 212).
344. The composition of claim 342 or 343, wherein the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 211).
345. The composition of any one of claims 342-344, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQADYEFGLIT (SEQ ID NO: 216).
346. The composition of any one of claims 342-345, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 215).
347. The composition of any one of claims 342-346, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 214).
348. The composition of any one of claims 342-347, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 211), a HC CDR2 sequence of SISPYSSYTSYADSVKG (SEQ ID NO: 212), a HC CDR3 sequence of YWYYMGMDY (SEQ ID NO: 213), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 214), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 215), and a LC CDR3 sequence of QQADYEFGLIT (SEQ ID NO: 216).
349. The composition of any one of claims 342-348, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVASISP YSSYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYMG MDYWGQGTLVTVSS (SEQ ID NO: 210).
350. The composition of any one of claims 342-349, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQADYEFGLITFGQGTKVEI KRTV (SEQ ID NO: 209).
351. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 221).
352. The composition of claim 351, wherein the VH comprises a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 220).
353. The composition of claim 351 or 352, wherein the VH comprises a HC CDR1 sequence of FSSYSIH (SEQ ID NO: 219).
354. The composition of any one of claims 351-353, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 224).
355. The composition of any one of claims 351-354, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 223).
356. The composition of any one of claims 351-355, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 222).
357. The composition of any one of claims 351-356, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSYSIH (SEQ ID NO: 219), a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 220), a HC CDR3 sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 221), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 222), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 223), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 224).
358. The composition of any one of claims 351-357, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSIHWVRQAPGKGLEWVAYISS SYGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYY HSQVEPPMAMDYWGQGTLVTVSS (SEQ ID NO: 218).
359. The composition of any one of claims 351-358, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEI KRTV (SEQ ID NO: 217).
360. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 229).
361. The composition of claim 360, wherein the VH comprises a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 228).
362. The composition of claim 360 or 361, wherein the VH comprises a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 227).
363. The composition of any one of claims 360-362, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 232).
364. The composition of any one of claims 360-363, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 231).
365. The composition of any one of claims 360-364, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 230).
366. The composition of any one of claims 360-365, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSYYSIH (SEQ ID NO: 227), a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 228), a HC CDR3 sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 229), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 230), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 231), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 232).
367. The composition of any one of claims 360-366, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYSIHWVRQAPGKGLEWVAYIS SSYGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYY HSQVEPPMAMDYWGQGTLVTVSS (SEQ ID NO: 226).
368. The composition of any one of claims 360-367, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEI KRTV (SEQ ID NO: 225).
369. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 237).
370. The composition of claim 369, wherein the VH comprises a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 236).
371. The composition of claim 369 or 370, wherein the VH comprises a HC CDR1 sequence of VSSSSIH (SEQ ID NO: 235).
372. The composition of any one of claims 369-371, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 240).
373. The composition of any one of claims 369-372, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 239).
374. The composition of any one of claims 369-373, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 238).
375. The composition of any one of claims 369-374, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSSSSIH (SEQ ID NO: 235), a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 236), a HC CDR3 sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 237), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 238), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 239), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 240).
376. The composition of any one of claims 369-375, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSSSIHWVRQAPGKGLEWVAYISS SYGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYY HSQVEPPMAMDYWGQGTLVTVSS (SEQ ID NO: 234).
377. The composition of any one of claims 369-376, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEI KRTV (SEQ ID NO: 233).
378. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 245).
379. The composition of claim 378, wherein the VH comprises a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 244).
380. The composition of claim 378 or 379, wherein the VH comprises a HC CDR1 sequence of VSYSSIH (SEQ ID NO: 243).
381. The composition of any one of claims 378-380, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSWWSYPLT (SEQ ID NO: 248).
382. The composition of any one of claims 378-381, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 247).
383. The composition of any one of claims 378-382, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 246).
384. The composition of any one of claims 378-383, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSYSSIH (SEQ ID NO: 243), a HC CDR2 sequence of YISSSYGYTSYADSVKG (SEQ ID NO: 244), a HC CDR3 sequence of SSRQYYHSQVEPPMAMDY (SEQ ID NO: 245), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 246), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 247), and a LC CDR3 sequence of QQSWWSYPLT (SEQ ID NO: 248).
385. The composition of any one of claims 378-384, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSYSSIHWVRQAPGKGLEWVAYIS SSYGYTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARSSRQYY HSQVEPPMAMDYWGQGTLVTVSS (SEQ ID NO: 242).
386. The composition of any one of claims 378-385, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSWWSYPLTFGQGTKVEI KRTV (SEQ ID NO: 241).
387. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 (HC CDR3) sequence of YWYYLGMDY (SEQ ID NO: 253).
388. The composition of claim 387, wherein the VH comprises a HC CDR2 sequence of YISPYSGYTYYADSVKG (SEQ ID NO: 252).
389. The composition of claim 387 or 388, wherein the VH comprises a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 251).
390. The composition of any one of claims 387-389, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQGSSSLLT (SEQ ID NO: 256).
391. The composition of any one of claims 387-390, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 255).
392. The composition of any one of claims 387-391, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 254).
393. The composition of any one of claims 387-392, wherein the antigen-binding domain comprises: a HC CDR1 sequence of VSSYSIH (SEQ ID NO: 251), a HC CDR2 sequence of YISPYSGYTYYADSVKG (SEQ ID NO: 252), a HC CDR3 sequence of YWYYLGMDY (SEQ ID NO: 253), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 254), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 255), and a LC CDR3 sequence of QQGSSSLLT (SEQ ID NO: 256).
394. The composition of any one of claims 387-393, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTVSSYSIHWVRQAPGKGLEWVAYIS PYSGYTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARYWYYL GMDYWGQGTLVTVSS (SEQ ID NO: 250).
395. The composition of any one of claims 387-394, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQGSSSLLTFGQGTKVEIKR TV (SEQ ID NO: 249).
396. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR3 comprising the amino acid sequence of LWASGLDY; and/or
(b) the VL comprises:
(i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
397. The composition of claim 396, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes.
398. The composition of claim 396 or 397, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes.
399. The composition of any one of claims 396-398, wherein the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
400. The composition of any one of claims 396-399, wherein the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
401. The composition of any one of claims 396-400, wherein the VH comprises:
(a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(c) a HC CDR3 comprising an amino acid sequence of LWASGLDY.
402. The composition of any one of claims 396-401, wherein the VL comprises:
(a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
403. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR3 comprising the amino acid sequence of SYYGFWQALWALDY; and/or
(b) the VL comprises:
(i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
404. The composition of claim 403, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes.
405. The composition of claim 403 or 404, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes.
406. The composition of any one of claims 403-405, wherein the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
407. The composition of any one of claims 403-406, wherein the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
408. The composition of any one of claims 403-407, wherein the VH comprises:
(a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(c) a HC CDR3 comprising an amino acid sequence of SYYGFWQALWALDY.
409. The composition of any one of claims 403-408, wherein the VL comprises:
(a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
410. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR3 comprising the amino acid sequence of GYYYPYYAMDY; and/or
(b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
411. The composition of claim 410, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes.
412. The composition of claim 410 or 411, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes.
413. The composition of any one of claims 410-412, wherein the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
414. The composition of any one of claims 410-413, wherein the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
415. The composition of any one of claims 410-414, wherein the VH comprises:
(a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(c) a HC CDR3 comprising an amino acid sequence of GYYYPYYAMDY.
416. The composition of any one of claims 410-415, wherein the VL comprises:
(a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
417. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises: (i) a HC CDR3 comprising the amino acid sequence of GYYYPYYAMDY; and/or
(b) the VL comprises:
(i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
418. The composition of claim 417, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes.
419. The composition of claim 417 or 418, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes.
420. The composition of any one of claims 417-419, wherein the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
421. The composition of any one of claims 417-420, wherein the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
422. The composition of any one of claims 417-421, wherein the VH comprises:
(a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(c) a HC CDR3 comprising an amino acid sequence of GYYYPYYAMDY.
423. The composition of any one of claims 417-422, wherein the VL comprises:
(a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
424. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR3 comprising the amino acid sequence of X1X2X3X4X5MDY, wherein Xi is F or G, X2 is Q or Y, X3 is W or G, X4 is Y or W, and X5 is A or G; and/or
(b) the VL comprises:
(i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
425. The composition of claim 424, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes.
426. The composition of claim 424 or 425, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes.
427. The composition of any one of claims 424-426, wherein the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
428. The composition of any one of claims 424-427, wherein the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
429. The composition of any one of claims 424-428, wherein the VH comprises:
(a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(c) a HC CDR3 comprising an amino acid sequence of X1X2X3X4X5MDY, wherein Xi is F or G, X2 is Q or Y, X3 is W or G, X4 is Y or W, and X5 is A or G.
430. The composition of any one of claims 424-429, wherein the VL comprises: (a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
431. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR3 comprising the amino acid sequence of SGYYSSHWYLQSWYQAMDY; and/or
(b) the VL comprises:
(i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
432. The composition of claim 431, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes.
433. The composition of claim 431 or 432, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes.
434. The composition of any one of claims 431-433, wherein the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
435. The composition of any one of claims 431-434, wherein the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
436. The composition of any one of claims 431-435, wherein the VH comprises:
(a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(c) a HC CDR3 comprising an amino acid sequence of SGYYSSHWYLQSWYQAMDY.
437. The composition of any one of claims 431-436, wherein the VL comprises:
(a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
438. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR3 comprising the amino acid sequence of SGYYSSHWYLQSWYQAMDY; and/or
(b) the VL comprises:
(i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
439. The composition of claim 438, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes.
440. The composition of claim 438 or 439, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes.
441. The composition of any one of claims 438-440, wherein the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
442. The composition of any one of claims 438-441, wherein the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
443. The composition of any one of claims 438-442, wherein the VH comprises:
(a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(c) a HC CDR3 comprising an amino acid sequence of SGYYSSHWYLQSWYQAMDY.
444. The composition of any one of claims 438-443, wherein the VL comprises:
(a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
445. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR3 comprising the amino acid sequence of HYSEKWWGWYTMYIDAMDY; and/or
(b) the VL comprises:
(i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
446. The composition of claim 445, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes.
447. The composition of claim 445 or 446, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes.
448. The composition of any one of claims 445-447, wherein the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
449. The composition of any one of claims 445-448, wherein the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
450. The composition of any one of claims 445-449, wherein the VH comprises:
(a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(c) a HC CDR3 comprising an amino acid sequence of HYSEKWWGWYTMYIDAMDY.
451. The composition of any one of claims 445-450, wherein the VL comprises:
(a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
452. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR3 comprising the amino acid sequence of X1WYYX2GMDY, wherein Xi is G or Y and X2 is M or L; and/or
(b) the VL comprises: (i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
453. The composition of claim 452, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes.
454. The composition of claim 452 or 453, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes.
455. The composition of any one of claims 452-454, wherein the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
456. The composition of any one of claims 452-455, wherein the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
457. The composition of any one of claims 452-456, wherein the VH comprises:
(a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(c) a HC CDR3 comprising an amino acid sequence of X1WYYX2GMDY, wherein Xi is G or Y and X2 is M or L.
458. The composition of any one of claims 452-457, wherein the VL comprises:
(a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein Xi is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
459. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein (a) the VH comprises:
(i) a HC CDR3 comprising the amino acid sequence of X1WYYX2GMDY, wherein Xi is G or Y and X2 is M or L; and/or
(b) the VL comprises:
(i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
460. The composition of claim 459, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes.
461. The composition of claim 459 or 460, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes.
462. The composition of any one of claims 459-461, wherein the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
463. The composition of any one of claims 459-462, wherein the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
464. The composition of any one of claims 459-463, wherein the VH comprises:
(a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(c) a HC CDR3 comprising an amino acid sequence of X1WYYX2GMDY, wherein Xi is G or Y and X2 is M or L.
465. The composition of any one of claims 459-464, wherein the VL comprises:
(a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or (c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6T, wherein Xi is G or A, X2 is S or Y, X3 is S or Y, X4 is G, D, or S, X5 is P or L, and Xe is I or L.
466. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR3 comprising the amino acid sequence of X1WYYX2GMDY, wherein Xi is G or Y and X2 is M or L; and/or
(b) the VL comprises:
(i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
467. The composition of claim 466, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes.
468. The composition of claim 466 or 467, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes.
469. The composition of any one of claims 466-468, wherein the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
470. The composition of any one of claims 466-469, wherein the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
471. The composition of any one of claims 466-470, wherein the VH comprises:
(a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes;
(b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(c) a HC CDR3 comprising an amino acid sequence of X1WYYX2GMDY, wherein Xi is G or Y and X2 is M or L.
472. The composition of any one of claims 466-471, wherein the VL comprises:
(a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4X5X6LX7T, wherein Xi is W, S, or A, X2 is N, W, S, or D, X3 is W, S, or Y, X4 is G, L, S, E or Y, X5 is W S, Y, E, or F, X6 is P, W, G, E, or Q, and X7 is I, F, or V.
473. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein
(a) the VH comprises:
(i) a HC CDR3 comprising the amino acid sequence of SSRQYYHSQVEPPMAMDY; and/or
(b) the VL comprises:
(i) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein XI is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
474. The composition of claim 473, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes.
475. The composition of claim 473 or 474, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes.
476. The composition of any one of claims 473-475, wherein the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
477. The composition of any one of claims 473-476, wherein the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes.
478. The composition of any one of claims 473-477, wherein the VH comprises:
(a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(c) a HC CDR3 comprising an amino acid sequence of
S SRQ YYHSQVEPPMAMD Y.
479. The composition of any one of claims 473-478, wherein the VL comprises:
(a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and/or
(c) a LC CDR3 comprising the amino acid sequence of QQX1X2X3X4YPX5T, wherein XI is W or S, X2 is K, W, or S, X3 is W, T, or A, X4 is S, W, or Y, and X5 is I, L, or V.
480. The composition of any one of claims 102-479, wherein the antigen-binding domain binds to a peptide conjugate/MHC complex, wherein the peptide conjugate of the peptide conjugate/MHC complex is a peptide covalently linked to a targeted covalent inhibitor or fragment thereof and a MHC.
481. The composition of claim 480, wherein the targeted covalent inhibitor is adagrasib, sotorasib, or divarasib.
482. The composition of claim 480, wherein the peptide conjugate/MHC complex is a first peptide conjugate/MHC complex comprising a first peptide conjugate comprising a first peptide and a first targeted covalent inhibitor or fragment thereof; and wherein the antigen-binding domain further binds to a second peptide conjugate/MHC complex comprising (a) a second peptide conjugate, wherein the second peptide conjugate of the second peptide conjugate/MHC complex is a second peptide covalently linked to a second targeted covalent inhibitor or fragment thereof; and (b) a second MHC.
483. The composition of claim 482, wherein the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 50 nM and (ii) the second peptide conjugate/MHC complex with a KD of at most about 50 nM.
484. The composition of claim 482, wherein the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 10 nM and (ii) the second peptide conjugate/MHC complex with a KD of at most about 10 nM.
485. The composition of claim 482, wherein the antigen-binding domain binds to (i) the first peptide conjugate/MHC complex with a dissociation constant (KD) of at most about 5 nM and (ii) the second peptide conjugate/MHC complex with a KD of at most about 5 nM.
486. The composition of any one of claims 482-485, wherein (i) the first targeted covalent inhibitor is adagrasib and the second targeted covalent inhibitor is sotorasib; (ii) the first targeted covalent inhibitor is adagrasib and the second targeted covalent inhibitor is divarasib; or (iii) the first targeted covalent inhibitor is divarasib and the second targeted covalent inhibitor is sotorasib.
487. The composition of any one of claims 480-486, wherein the antigen-binding domain does not bind to a free first targeted covalent inhibitor with a dissociation constant (KD) of less than 200 nM.
488. The composition of claim 487, wherein the antigen-binding domain binds to the first peptide conjugate/MHC complex with a greater affinity than to the first peptide or the free first targeted covalent inhibitor.
489. The composition of claim 487 or 488, wherein the affinity of the antigen-binding domain for the first peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the antigen-binding domain for the first peptide or the free first targeted covalent inhibitor.
490. The composition of any one of claims 482-489, wherein the antigen-binding domain does not detectably bind to a complex of the first peptide with a MHC, wherein the first peptide is not covalently bound to the first targeted covalent inhibitor or fragment thereof.
491. The composition of any one of claims 487-490, wherein the antigen-binding domain does not detectably bind to the free first targeted covalent inhibitor.
492. The composition of any one of claims 482-491, wherein the antigen-binding domain binds to a free first peptide conjugate with a dissociation constant KD that is at least 2.5 times more than a KD of the antibody or the antigen-binding fragment to the first peptide conjugate/MHC complex.
493. The composition of any one of claims 482-492, wherein the antigen-binding domain does not bind to a free second targeted covalent inhibitor with a dissociation constant (KD) of less than 200 nM.
494. The composition of claim 493, wherein the antigen-binding domain binds to the second peptide conjugate/MHC complex with a greater affinity than to the second peptide or the free second targeted covalent inhibitor.
495. The composition of claim 493 or 494, wherein the affinity of the antigen-binding domain for the second peptide conjugate/MHC complex is 100-10,000 times greater than the affinity of the antigen-binding domain for the second peptide or the free second targeted covalent inhibitor.
496. The composition of any one of claims 482-495, wherein the antigen-binding domain does not detectably bind to a complex of the second peptide with the second MHC, wherein the second peptide is not covalently bound to the second targeted covalent inhibitor or fragment thereof.
497. The composition of any one of claims 493-496, wherein the antigen-binding domain does not detectably bind to the free second targeted covalent inhibitor.
498. The composition of any one of claims 482-497, wherein the antigen-binding domain binds to a free second peptide conjugate with a dissociation constant ( o) that is at least 2.5 times more than a Ko of the antibody or the antigen-binding fragment to the second peptide conjugate/MHC complex.
499. The composition of any one of claims 482-498, wherein the first peptide and the second peptide comprise the same amino acid sequence.
500. The composition of any one of claims 482-499, wherein the first peptide and the second peptide consist of the same amino acid sequence.
501. The composition of claim 499 or 500, wherein the first peptide or the second peptide comprises the amino acid sequence of VVVGACGVGK.
502. The composition of any one of claims 482-498, wherein the first peptide and second peptide comprise a different amino acid sequence.
503. The composition of claim 502, wherein the first peptide or the second peptide comprises an amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV.
504. The composition of any one of claims 482-503, wherein the MHC is an HLA, and wherein the HLA is HLA-A*02:01, HLA-A*03:01, or HLA-A*11 :01.
505. The composition of any one of claims 482-504, wherein the first peptide or the second peptide is or is derived from KRAS.
506. The composition of any one of claims 482-505, wherein the first peptide or the second peptide comprises a segment of KRASG12C, KRASG12D, KRASG12R, or KRASG12S.
507. The composition of any one of claims 482-506, wherein the first peptide conjugate is formed by the covalent reaction of a free first targeted covalent inhibitor with a KRASG12C peptide, a KRASG12D peptide, a KRASG12S peptide, or a KRASG12R peptide.
508. The composition of any one of claims 482-507, wherein the second peptide conjugate is formed by the covalent reaction of a second free targeted covalent inhibitor with a KRASG12C peptide, a KRASG12D peptide, a KRASG12S peptide, or a KRASG12R peptide.
509. The composition of claim 507 or 508, wherein the free first targeted covalent inhibitor or the second free targeted covalent inhibitor is sotorasib, adagrasib, or divarasib.
510. The composition of any one of claims 482-509, wherein the first peptide or the second peptide comprises the amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV.
511. The composition of any one of claims 102-479, wherein the antigen-binding domain binds to
(a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01;
(b) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01; or
(c) a first peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
512. The composition of any one of claims 102-479, wherein the antigen-binding domain binds to
(a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01;
(b) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01; or
(c) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
513. The composition of any one of claims 102-479, wherein the antigen-binding domain binds to
(a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HL A- A* 11 :01;
(b) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HL A- A* 11 :01; or
(c) a first peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
514. The composition of any one of claims 102-479, wherein the antigen-binding domain binds to
(a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HL A- A* 11 :01;
(b) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01; or
(c) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
515. The composition of any one of claims 102-479, wherein the antigen-binding domain binds to
(a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HL A- A* 11 :01;
(b) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HL A- A* 11 :01; or
(c) a first peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
516. The composition of any one of claims 102-479, wherein the antigen-binding domain binds to
(a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HL A- A* 11 :01;
(b) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01; or
(c) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
517. The composition of any one of claims 102-479, wherein the antigen-binding domain binds to
(a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01;
(b) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01; or
(c) a first peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
518. The composition of any one of claims 102-479, wherein the antigen-binding domain binds to
(a) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*03:01;
(b) a first peptide conjugate/MHC complex comprising VVVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01; or
(c) a first peptide conjugate/MHC complex comprising VVGACGVGK conjugated to a first targeted covalent inhibitor or fragment thereof presented by HLA-A* 11 :01 and a second peptide conjugate/MHC complex comprising KLVVVGACGV conjugated to a second targeted covalent inhibitor or fragment thereof presented by HLA-A*02:01.
519. The composition of any one of claims 511-518, wherein the first targeted covalent inhibitor or the second targeted covalent inhibitor is sotorasib, adagrasib, or divarasib.
520. The composition of any one of claims 102-519, wherein the VH is linked to the VL via a linker sequence.
521. The composition of claim 520, wherein the linker sequence comprises (G4S)n or (S4G)n, wherein n is any integer from 1 to 10.
522. The composition of claim 520 or 521, wherein the linker comprises the glycine- serine-alanine linker G4SA3 or a glycine-serine linker (G4S)4.
523. The composition of any one of claims 102-522, wherein the polypeptide is an intact antibody, a bispecific antibody, a multispecific antibody, an antigen-binding (Fab) fragment, an Fab’ fragment, an (Fab’)2 fragment, an Fd, an Fv, a dAb, a single domain fragment or single monomeric variable antibody domain, a single-chain Diabody (scDb), a diabody (Db), a dual-affinity retargeting (DART) molecule, a single-chain variable fragment (scFv), a Bi-specific T-cell engager (BiTE), bispecific killer cell engager (BiKE), CrossMab, a tri-specific binding partner, a chimeric antigen receptor (CAR), a camelid antibody, a monobody, a DARPin, an anticalin, an affibody, or an affimer.
524. The composition of any one of claims 102-522, wherein the polypeptide comprises a monobody, a DARPin, an anticalin, an affibody, or an affimer.
525. The composition of any one of claims 102-524, wherein the polypeptide comprises a polypeptide chain comprising the antigen-binding domain.
526. The composition of claim 525, wherein the polypeptide chain further comprises a cytokine or fragment thereof.
527. The composition of claim 526, wherein cytokine comprises IL-2, IL-7, IL-15, IL-12, IL- 18, IL-21, or an interferon (IFN).
528. A pharmaceutical composition comprising a polypeptide of any one of claims 170- 527, and a pharmaceutically acceptable carrier, excipient, adjuvant or diluent.
529. A method of treating cancer in a subject in need thereof wherein the subject has been treated with a free targeted covalent inhibitor, the method comprising administering to the subject the polypeptide of any one of claims 170-527 or the pharmaceutical composition of claim 528.
530. The method of claim 529, wherein the free targeted covalent inhibitor is sotorasib, adagrasib, or divarasib.
531. The method of claim 529 or 530, wherein the subject is refractory to a treatment with the free targeted covalent inhibitor.
532. A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject the polypeptide of any one of claims 170-527 or the pharmaceutical composition of claim 528.
533. The method of claim 532, wherein the method further comprises administering simultaneously a small molecule drug.
534. A method of administering to the subject in need thereof the polypeptide of any one of claims 170-527 or the pharmaceutical composition of claim 528, and a free targeted covalent inhibitor, wherein the free targeted covalent inhibitor is optionally sotorasib, adagrasib, or divarasib.
535. The method of claim 534, wherein the polypeptide of any one of claims 170-527 or the pharmaceutical composition of claim 528 is administered after administration of the free targeted covalent inhibitor or simultaneously with the free targeted covalent inhibitor.
536. A method of manufacturing a T-cell receptor (TCR) that recognizes the first peptide conjugate/MHC complex or the second peptide conjugate/MHC complex of any one of claims 482-510, the method comprising:
(a) a plurality of candidate TCRs with the first peptide conjugate/MHC complex or the second peptide conjugate/MHC complex, and
(b) identifying at least one TCR that binds to the first peptide conjugate/MHC complex or the second peptide conjugate/MHC complex.
537. The method of claim 536, wherein identifying in (b) further comprises selecting or isolating the at least one TCR.
538. The method of claim 536 or 537, wherein the plurality of candidate TCRs is a plurality of soluble TCRs or a plurality of TCRs expressed on cell surface of a plurality of cells.
539. The method of claim 538, wherein the plurality of candidate TCRs is the plurality of TCRs expressed on cell surface of the plurality of cells, and identifying in (b) comprises isolating or selecting a cell comprising the at least one TCR based on an activation marker of the cell.
540. The method of claim 539, wherein the activation marker is a T cell activation marker.
541. The method of claim 540, wherein the T cell activation marker is CD26, CD27, CD28, CD30, CD154, CD40L, CD134. CD25, CD44, CD69, CD137, PD-1, or KLRG1
542. A T-cell receptor (TCR) comprising the at least one TCR of (b) of any one of claims 536-541.
543. The TCR of claim 542, wherein the TCR is a soluble TCR.
544. The TCR of claim 542 or 543, wherein the TCR is a bispecific TCR.
545. The composition of any one of claims 1-90 and 482-510, wherein the first peptide conjugate/MHC complex is different from the second peptide conjugate/MHC complex, wherein each of the first and second peptide conjugate/MHC complexes independently comprises:
(a) a different peptide selected from the group consisting of peptides comprising the formula
X#X#+iX#+2X#+3X#+4(X#+5)X#+6X#+7X#+sX#+9, and
X#X#+ 1 X#+2X#+3 (X#+4)X#+sX#+eX#+7X#+8X#+9, and X#X#+iX#+2(X#+3)X#+4X#+sX#+6X#+7X#+8X#+9, and X#X#+i(X#+2)X#+3X#+4X#+sX#+6X#+7X#+8X#+9, and X#-iX#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9, and X#+lX#+2X#+3X#+4(X#+5)X#+6X#+7X#+8X#+9, wherein the peptide conjugate of the first and second peptide conjugate/MHC complex is formed by the covalent reaction of the first targeted covalent inhibitor, second targeted covalent inhibitor or fragment thereof with the residue in parenthesis; and
(b) the same MHC; wherein the polypeptide binds to the first peptide conjugate/MHC complex with a KD that is at most 100 nM and binds to the second peptide conjugate/MHC complex with an KD that is at most 100 nM, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to a background- subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to the biolayer interferometry analysis, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to a background-subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to the background-subtracted biolayer interferometry analysis, and wherein the reference binding signal is the maximum binding signal of the polypeptide binding to X#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9.
546. The composition of claim 545, wherein the formula X#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9 is the formula X7X8X9X10X11X12X13X14X15X16, and wherein X12 is covalently linked to the first targeted covalent inhibitor, second targeted covalent inhibitor or fragment thereof.
547. The composition of any one of claims 1-90 and 482-510, wherein the polypeptide binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex, wherein each of the first and the second peptide conjugate/MHC complex comprises:
(a) a peptide comprising the formula X7X8X9X10X11X12X13X14X15X16; and
(b) the same MHC; wherein the first peptide conjugate of the first peptide conjugate/MHC complex is formed by the covalent reaction of the first targeted covalent inhibitor or fragment thereof with X12 of the peptide, and wherein the second peptide conjugate of the second peptide conjugate/MHC complex is formed by the covalent reaction of the second targeted covalent inhibitor or fragment thereof with X13 of the peptide, wherein the second targeted covalent inhibitor is the same as the first targeted covalent inhibitor; and wherein the polypeptide binds to the second peptide conjugate/MHC complex with a KD that is at most 100,000-fold higher than the KD of the polypeptide to the first peptide conjugate/MHC complex, or wherein a maximum wavelength shift of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum wavelength shift of the polypeptide binding to the first peptide conjugate/MHC complex, or wherein a maximum binding signal of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum binding signal of the polypeptide binding to the first peptide conjugate/MHC complex.
548. The composition of any one of claims 1-90 and 482-510, wherein the polypeptide binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex, wherein each of the first and the second peptide conjugate/MHC complex comprises:
(a) a peptide comprising the formula X7X8X9X10X11X12X13X14X15X16; and
(b) the same MHC; wherein the first peptide conjugate of the first peptide conjugate/MHC complex is formed by the covalent reaction of the first targeted covalent inhibitor or fragment thereof with a residue selected from the group consisting of X9, X10, X11, and X12, and wherein the second peptide conjugate of the second peptide conjugate/MHC complex is formed by the covalent reaction of the second targeted covalent inhibitor or fragment thereof with a residue selected from the group consisting of X13, X14, and X15, wherein the second targeted covalent inhibitor is the same as the first targeted covalent inhibitor; wherein the polypeptide binds to the second peptide conjugate/MHC complex with a KD that is at most 100,000 fold higher that the KD of the polypeptide to the first peptide conjugate/MHC complex, or wherein a maximum wavelength shift of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum wavelength shift of the polypeptide binding to the first peptide conjugate/MHC complex, or wherein a maximum binding signal of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum binding signal of the polypeptide binding to the first peptide conjugate/MHC complex.
549. The composition of any one of claims 1-90 and 482-510, wherein the polypeptide binds to the first peptide conjugate/MHC complex and the second peptide conjugate/MHC complex, wherein each of the first and the second peptide conjugate/MHC complex comprises: a peptide comprising the formula A-B-C, where A comprises no more than three residues having an N-terminal anchor residue, B comprises at least four but no more than seven residues having a residue covalently linked to the first targeted covalent inhibitor or fragment thereof or the second targeted covalent inhibitor or fragment thereof, and C comprises no more than three residues having a C-terminal anchor residue, wherein the N-terminal anchor residues and the C-terminal anchor residue bind to an MHC; and the same MHC; wherein the residue covalently linked to the first targeted covalent inhibitor or fragment thereof of the peptide of the first peptide conjugate/MHC complex is different from the residue covalently linked to the second targeted covalent inhibitor or fragment thereof of the peptide of the second peptide conjugate/MHC complex; and wherein the polypeptide binds to the first peptide conjugate/MHC complex with a KD that is at most 100 nM and binds to the second peptide conjugate/MHC complex with a KD that is at most 100 nM, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to a background- subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to the background- subtracted biolayer interferometry analysis, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to a background-subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to the background-subtracted biolayer interferometry analysis, and wherein the reference binding signal is the maximum binding signal of the polypeptide binding to X#X#+iX#+2X#+3X#+4(X#+5)X#+6X#+7X#+sX#+9.
550. The composition of claim 549, wherein A comprises two residues X?Xs.
551. The composition of claim 549 or 550, wherein B comprises seven residues X9X10X11X12X13X14X15.
552. The composition of any one of claims 549-551, wherein C comprises one residue Xi6.
553. The composition of any one of claims 549-552, wherein the formula A-B-C is the formula X7X8X9X10X11X12X13X14X15X16.
554. The composition of claim 553, wherein the first peptide conjugate of the first peptide conjugate/MHC complex is formed by the covalent reaction of the first targeted covalent inhibitor or fragment thereof with X12 of the peptide.
555. The composition of claim 553 or 554, wherein the second peptide conjugate of the second peptide conjugate/MHC complex is formed by the covalent reaction of the second targeted covalent inhibitor or fragment thereof with X13 of the peptide, wherein the second targeted covalent inhibitor is the same as the first targeted covalent inhibitor.
556. The composition of claim 555, wherein the polypeptide binds to the second peptide conjugate/MHC complex with a KD that is at most 100,000 fold higher than the KD of the polypeptide to the first peptide conjugate/MHC complex.
557. The composition of any one of claims 551-556, wherein the polypeptide binds to the second peptide conjugate/MHC complex with an KD that is at most 100,000 fold higher than the KD of the polypeptide to the first peptide conjugate/MHC complex, and wherein the residue covalently linked to the first targeted covalent inhibitor or fragment thereof of the peptide of the first peptide conjugate/MHC complex is selected from the group of consisting of X9, X10, X11, and X12, and the residue covalently linked to the second targeted covalent inhibitor or fragment thereof of the peptide of the second peptide conjugate/MHC complex is X13, X14, and X15.
558. The composition of any one of claims 544-557, wherein the peptide is a RAS peptide.
559. The composition of claim 558, wherein the RAS peptide comprises a mutation.
560. The composition of claim 559, wherein the mutation is G12C or G13C.
561. The composition of any one of claims 558-560, wherein the RAS peptide comprises a sequence selected from the group consisting of VVVGACGVGK, VVGACGVGK, and KLVVVGACGV.
562. The composition of any one of claims 558-560, wherein the RAS peptide comprises a sequence selected from the group consisting of VVVGAGCVGK, VVGAGCVGK, or KLVVVGAGCV.
563. The composition of any one of claims 544-562, wherein the same MHC is selected from the group consisting of HLA-A*03:01, HLA-A* 11:01, HLA-A*02:01, HLA- A*68:01, HLA-A*31:01, HLA-A*30:01, HLA-A*33:03, HLA-A*33:01, HLA- A*74:01, HLA-A*34:02, HLA-A*66:01, HLA-A*68:02, HLA-A*02:05, HLA- A*02:02, and HLA-A*02:06.
564. The composition of any one of claims 544-563, wherein the targeted covalent inhibitor or fragment thereof comprises sotorasib.
565. The composition of any one of claims 546-548 and 553-564, wherein the peptide is a RAS peptide, and wherein X12 is G12C mutation, and X13 is G13C mutation.
566. The composition of claim 565, wherein the polypeptide binds to the first peptide conjugate/MHC complex having a Cys of the G12C mutation covalently linked to the first targeted covalent inhibitor or fragment thereof with a KD that is at most 100 nM and binds to the second peptide conjugate/MHC complex having a Cys of the G13C mutation covalently linked to the second targeted covalent inhibitor or fragment thereof with an KD that is most 100 nM, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to a background- subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to the background- subtracted biolayer interferometry analysis, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to a background-subtracted biolayer interferometry analysis and wherein when a polypeptide is contact to the second peptide conjugate/MHC complex a maximum binding signal of at least 0.1 -fold of a reference binding signal is observed according to a background-subtracted biolayer interferometry analysis, and wherein the reference binding signal is the maximum binding signal of the polypeptide binding to X#X#+iX#+2X#+3X#+4X#+sX#+6X#+7X#+8X#+9.
567. The composition of claim 565, wherein the polypeptide binds to the second peptide conjugate/MHC complex having a Cys of the G13C mutation covalently linked to the second targeted covalent inhibitor or fragment thereof with a KD that is at most 100,000 fold higher that the KD of the polypeptide to the first peptide conjugate/MHC complex having a Cys of the G12C mutation covalently linked to the first targeted covalent inhibitor or fragment thereof, or wherein a maximum wavelength shift of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum wavelength shift of the polypeptide binding to the first peptide conjugate/MHC complex, or wherein a maximum binding signal of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum binding signal of the polypeptide binding to the first peptide conjugate/MHC complex.
568. A composition comprising: a polypeptide that binds to a first peptide conjugate/MHC complex and a second peptide conjugate/MHC complex different from the first peptide conjugate/MHC complex, wherein each of the first and second peptide conjugate/MHC complexes independently comprises:
(a) a different peptide selected from the group consisting of peptides comprising the formula
X#X#+iX#+2X#+3X#+4(X#+5)X#+6X#+7X#+sX#+9, and
X#X#+ 1 X#+2X#+3 (X#+4)X#+sX#+eX#+7X#+8X#+9, and X#X#+iX#+2(X#+3)X#+4X#+sX#+6X#+7X#+8X#+9, and X#X#+i(X#+2)X#+3X#+4X#+sX#+6X#+7X#+8X#+9, and X#-iX#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9, and X#+lX#+2X#+3X#+4(X#+5)X#+6X#+7X#+8X#+9, wherein the peptide conjugate of the first and second peptide conjugate/MHC complex is formed by the covalent reaction of a targeted covalent inhibitor or fragment thereof with the residue in parenthesis; and
(b) the same MHC; wherein the polypeptide binds to the first peptide conjugate/MHC complex with an KD that is at most 100 nM and binds to the second peptide conjugate/MHC complex with an KD that is at most 100 nM, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to a background- subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to the background- subtracted biolayer interferometry analysis, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum binding signal of at least O.l-fold of a reference binding signal is observed according to a background-subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum binding signal of at least O.l-fold of a reference binding signal is observed according to the background-subtracted biolayer interferometry analysis, and wherein the reference binding signal is the maximum binding signal of the polypeptide binding to X#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9.
569. The composition of claim 568, wherein each of the first and second peptide conjugate/MHC complexes independently comprises: a different peptide selected from the group consisting of peptides comprising the formula X#X#+iX#+2X#+3X#+4(X#+s)X#+6X#+7X#+8X#+9, and
X#X#+ 1 X#+2X#+3 (X#+4)X#+sX#+eX#+7X#+8X#+9, and X#X#+iX#+2(X#+3)X#+4X#+sX#+6X#+7X#+8X#+9, and X#X#+l(X#+2)X#+3X#+4X#+5X#+6X#+7X#+8X#+9.
570. The composition of claim 568, wherein each of the first and second peptide conjugate/MHC complexes independently comprises: a different peptide with an amino acid sequence selected from the group consisting of: VVVGACGVGK, VVVGCGGVGK, VVVCAGGVGK, and VVCGAGGVGK.
571. A composition comprising a polypeptide that binds to a first peptide conjugate/MHC complex and a second peptide conjugate/MHC complex, wherein each of the first and the second peptide conjugate/MHC complex comprises:
(a) a peptide comprising the formula X7X8X9X10X11X12X13X14X15X16; and
(b) the same MHC; wherein a first peptide conjugate of the first peptide conjugate/MHC complex is formed by the covalent reaction of a targeted covalent inhibitor or fragment thereof with X12 of the peptide, and wherein a second peptide conjugate of the second peptide conjugate/MHC complex is formed by the covalent reaction of the same targeted covalent inhibitor or fragment thereof with X13 of the peptide; wherein a polypeptide binds to the second peptide conjugate/MHC complex with a KD that is at most 100,000 fold higher than the KD of the polypeptide to the first peptide conjugate/MHC complex, or wherein a maximum wavelength shift of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum wavelength shift of the polypeptide binding to the first peptide conjugate/MHC complex, or wherein a maximum binding signal of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum binding signal of the polypeptide binding to the first peptide conjugate/MHC complex.
572. A composition comprising a polypeptide that binds to a first peptide conjugate/MHC complex and a second peptide conjugate/MHC complex, wherein each of the first and the second peptide conjugate/MHC complex comprises:
(a) a peptide comprising the formula X7X8X9X10X11X12X13X14X15X16; and
(b) the same MHC; wherein a first peptide conjugate of the first peptide conjugate/MHC complex is formed by the covalent reaction of a targeted covalent inhibitor or fragment thereof with a residue selected from the group consisting of X9, X10, X11, and X12, and wherein a second peptide conjugate of the second peptide conjugate/MHC complex is formed by the covalent reaction of the same targeted covalent inhibitor or fragment thereof with a residue selected from the group consisting of X13, X14, and X15; wherein a polypeptide binds to the second peptide conjugate/MHC com-plex with a KD that is at most 100,000 fold higher that the KD of the polypeptide to the first peptide conjugate/MHC complex, or wherein a maximum wavelength shift of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum wavelength shift of the polypeptide binding to the first peptide conjugate/MHC complex, or wherein a maximum binding signal of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum binding signal of the polypeptide binding to the first peptide conjugate/MHC complex.
573. A composition comprising a polypeptide that binds to a first peptide conjugate/MHC complex and a second peptide conjugate/MHC complex, wherein each of the first and the second peptide conjugate/MHC complex comprises: a peptide comprising the formula A-B-C, where A comprises no more than three residues having an N-terminal anchor residue, B comprises at least four but no more than seven residues having a residue covalently linked to a targeted covalent inhibitor or fragment thereof, and C comprises no more than three residues having a C-terminal anchor residue, wherein the N-terminal anchor residues and the C-terminal anchor residue bind to an MHC; and the same MHC; wherein the residue covalently linked to a targeted covalent inhibitor or fragment thereof of the peptide of the first peptide conjugate/MHC complex is different from the residue covalently linked to a targeted covalent inhibitor or fragment thereof of the peptide of the second peptide conjugate/MHC complex; and wherein a polypeptide binds to the first peptide conjugate/MHC complex with a KD that is at most 100 nM and binds to the second peptide conjugate/MHC complex with a KD that is at most 100 nM, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to a background- subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to the background- subtracted biolayer interferometry analysis, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to a background-subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to the background-subtracted biolayer interferometry analysis, and wherein the reference binding signal is the maximum binding signal of the polypeptide binding to X#X#+iX#+2X#+3X#+4(X#+5)X#+6X#+7X#+sX#+9.
574. The composition of claim 573, wherein A comprises two residues X?Xs.
575. The composition of claim 573 or 574, wherein B comprises seven residues X9X10X11X12X13X14X15.
576. The composition of any one of claims 573-575, wherein C comprises one residue X16.
577. The composition of any one of claims 573-576, wherein the formula A-B-C is the formula X7X8X9X10X11X12X13X14X15X16.
578. The composition of claim 577, wherein a first peptide conjugate of the first peptide conjugate/MHC complex is formed by the covalent reaction of the targeted covalent inhibitor or fragment thereof with X12 of the peptide.
579. The composition of claim 577 or 578, wherein a second peptide conjugate of the second peptide conjugate/MHC complex is formed by the covalent reaction of the same targeted covalent inhibitor or fragment thereof with X13 of the peptide.
580. The composition of claim 579, wherein the polypeptide binds to the second peptide conjugate/MHC complex with a KD that is at most 100,000 fold higher than the KD of the polypeptide to the first peptide conjugate/MHC complex.
581. The composition of any one of claims 575-580, wherein the polypeptide binds to the second peptide conjugate/MHC complex with an KD that is at most 100,000 fold higher than the KD of the polypeptide to the first peptide conjugate/MHC complex, and wherein the residue covalently linked to the targeted covalent inhibitor or fragment thereof of the peptide of the first peptide conjugate/MHC complex is selected from the group of consisting of X9, X10, X11, and X12, and the residue covalently linked to the targeted covalent inhibitor or fragment thereof of the peptide of the second peptide conjugate/MHC complex is X13, X14, and X15.
582. The composition of any one of claims 568-581, wherein the peptide is a RAS peptide.
583. The composition of claim 582, wherein the RAS peptide comprises a mutation.
584. The composition of claim 583, wherein the mutation is G12C or G13C.
585. The composition of any one of claims 582-584, wherein the RAS peptide comprises a sequence selected from the group consisting of VVVGACGVGK, VVGACGVGK, and KLVVVGACGV.
586. The composition of any one of claims 582-584, wherein the RAS peptide comprises a sequence selected from the group consisting of VVVGAGCVGK, VVGAGCVGK, or KLVVVGAGCV.
587. The composition of any one of claims 568-586, wherein the same MHC is selected from the group consisting of HL A-A* 03:01, HLA-A* 11:01, HLA-A*02:01, HLA-, HLA-A*68:01, HLA-A*31:01, HLA-A*30:01, HLA-A*33:03, HLA-A*33:01, HLA- A*74:01, HLA-A*34:02, HLA-A*66:01, HLA-A*68:02, HLA-A*02:05, HLA- A*02:02, and HLA-A*02:06.
588. The composition of any one of claims 568-587, wherein the targeted covalent inhibitor or fragment thereof comprises sotorasib.
589. The composition of any one of claims 570-573 and 577-588, wherein the peptide is a RAS peptide, and wherein X12 is G12C mutation, and X13 is G13C mutation.
590. The composition of claim 589, wherein the polypeptide binds to the first peptide conjugate/MHC complex having a Cys of the G12C mutation covalently linked to the targeted covalent inhibitor or fragment thereof with an KD that is at most 100 nM and binds to the second peptide conjugate/MHC complex having a Cys of the G13C mutation covalently linked to the targeted covalent inhibitor or fragment thereof with an KD that is at most 100 nM, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to a background- subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum wavelength shift of at least 0.1 nm is observed according to the background- subtracted biolayer interferometry analysis, or wherein when the polypeptide is contacted to the first peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to a background-subtracted biolayer interferometry analysis and wherein when the polypeptide is contacted to the second peptide conjugate/MHC complex a maximum binding signal of at least 0.1-fold of a reference binding signal is observed according to the background-subtracted biolayer interferometry analysis, and wherein the reference binding signal is the maximum binding signal of the polypeptide binding to X X iX :X >X 'X iX X sX
591. The composition of claim 589, wherein the polypeptide binds to the second peptide conjugate/MHC complex having a Cys of the G13C mutation covalently linked to the targeted covalent inhibitor or fragment thereof with a KD that is at most 100,000 fold higher that the KD of the polypeptide to the first peptide conjugate/MHC complex having a Cys of the G12C mutation covalently linked to the targeted covalent inhibitor or fragment thereof, or wherein a maximum wavelength shift of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum wavelength shift of the polypeptide binding to the first peptide conjugate/MHC complex, or wherein a maximum binding signal of the polypeptide binding to the second peptide conjugate/MHC complex is at least 10-fold less than a maximum binding signal of the polypeptide binding to the first peptide conjugate/MHC complex.
592. The polynucleotide of claim 91 or 92, wherein the polynucleotide is a modified polynucleotide.
593. The composition of claim 525, wherein the polypeptide chain further comprises an agonist molecule.
594. The composition of claim 593, wherein the agonist molecule is a CD28 agonist or a 4- 1BB agonist.
595. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) and/or a light chain variable region (VL), wherein:
(a) the VH comprises:
(i) a HC CDR3 comprising the amino acid sequence of FX2X3X4AMDY, wherein X2 is Y, L, T, H, E, A, or R; X3 is D, V, L, E, H, T or R; and X4 is L or Y (SEQ ID NO: 267); and/or
(b) the VL comprises:
(i) a LC CDR3 comprising the amino acid sequence of QQX3SWLX7WX9X10T, wherein X3 is A or S, X7 is Y or H, X9 is L, K, V, Y or I, and X10 is L, V, or I (SEQ ID NO: 268).
596. The composition of claim 595, wherein the VH comprises: a HC CDR3 comprising the amino acid sequence of FX2X3X4AMDY, wherein X2 is Q, Y, L, T, H, E, A, or R; X3 is W, D, V, L, E, H, T or R; and X4 is L or Y.
597. The composition of claim 595 or 596, wherein the VH comprises a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes.
598. The composition of any one of claims 595-597, wherein the VH comprises a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes.
599. The composition of any one of claims 595-598, wherein the VL comprises a LC CDR1 comprising an amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes.
600. The composition of any one of claims 595-599, wherein the VL comprises a LC CDR2 comprising an amino acid sequence of SASSLYS, or a variant thereof comprising 1-3 amino acid changes.
601. The composition of any one of claims 595-600, wherein the VH comprises:
(a) a HC CDR1 comprising an amino acid sequence of FSSSSIH, or a variant thereof comprising 1-3 amino acid changes; (b) a HC CDR2 comprising an amino acid sequence of SISSYYGSTYYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and/or
(c) a HC CDR3 comprising an amino acid sequence of FX2X3X4AMDY, wherein X2 is Y, L, T, H, E, A; X3 is D, V, L, E, H, T or R; and X4 is L or Y.
602. The composition of any one of claims 595-601, wherein the VL comprises:
(a) a LC CDR1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;
(b) a LC CDR2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-3 amino acid changes, and/or
(c) a LC CDR3 comprising the amino acid sequence of QQX3SWLX7WX9X10T, wherein X3 is A or S, X7 is Y or H, Xs is L, K, V, Y or I, and X9 is L, V, or I.
603. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FYDLAMDY (SEQ ID NO.: 273).
604. The composition of claim 603, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 272).
605. The composition of claim 603 or 604, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 271).
606. The composition of any one of claims 603-605, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWLIT (SEQ ID NO.: 276).
607. The composition of claim 606, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 275).
608. The composition of claim 606 or 607, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 274).
609. The composition of any one of claims 603-608, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 271), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 272), a HC CDR3 sequence of FYDLAMDY (SEQ ID NO: 273), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 274), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 275), and a LC CDR3 sequence of QQASWLYWLIT (SEQ ID NO: 276).
610. The composition of any one of claims 603-609, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMD YWGQGTLVTVSS (SEQ ID NO.: 270).
611. The composition of any one of claims 606-610, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKV EIK (SEQ ID NO.: 269).
612. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FLDLAMDY (SEQ ID NO.: 281).
613. The composition of claim 612, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 280).
614. The composition of claim 612 or 613, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 279).
615. The composition of any one of claims 612-614, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWKIT (SEQ ID NO.: 284).
616. The composition of claim 615, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 283).
617. The composition of claim 615 or 616, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 282).
618. The composition of any one of claims 612-617, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 279), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 280), a HC CDR3 sequence of FLDLAMDY (SEQ ID NO: 281), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 282), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 283), and a LC CDR3 sequence of QQASWLYWKIT (SEQ ID NO: 284).
619. The composition of any one of claims 612-618, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFLDLAMD YWGQGTLVTVSS (SEQ ID NO.: 278).
620. The composition of any one of claims 615-619, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKITFGQGTKV EIK (SEQ ID NO.: 277).
621. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTVLAMDY (SEQ ID NO.: 289).
622. The composition of claim 621, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 288).
623. The composition of claim 621 or 622, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 287).
624. The composition of any one of claims 621-623, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWLIT (SEQ ID NO.: 292).
625. The composition of claim 624, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 291).
626. The composition of claim 624 or 625, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 290).
627. The composition of any one of claims 621-626, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 287), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 288), a HC CDR3 sequence of FTVLAMDY (SEQ ID NO.: 289), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 290), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 291), and a LC CDR3 sequence of QQASWLYWLIT (SEQ ID NO.: 292).
628. The composition of any one of claims 621-627, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMD YWGQGTLVTVSS (SEQ ID NO.: 286).
629. The composition of any one of claims 624-628, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKV EIK (SEQ ID NO.: 285).
630. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTVLAMDY (SEQ ID NO.: 297).
631. The composition of claim 630, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 296).
632. The composition of claim 630 or 631, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 295).
633. The composition of any one of claims 630-632, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWKVT (SEQ ID NO.: 300).
634. The composition of claim 633, wherein the VL comprises a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 299).
635. The composition of claim 633 or 634, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 298).
636. The composition of any one of claims 630-635, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 295), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 296), a HC CDR3 sequence of FTVLAMDY (SEQ ID NO.: 297), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 297), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 298), and a LC CDR3 sequence of QQASWLYWKVT (SEQ ID NO.: 300).
637. The composition of any one of claims 630-636, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMD YWGQGTLVTVSS (SEQ ID NO.: 294).
638. The composition of any one of claims 633-637, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKVTFGQGTKV EIK (SEQ ID NO.: 293).
639. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTVLAMDY (SEQ ID NO.: 305).
640. The composition of claim 639, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 304).
641. The composition of claim 639 or 640, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 303).
642. The composition of any one of claims 639-641, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWVIT (SEQ ID NO.: 308).
643. The composition of claim 642, wherein the VL comprises a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 307).
644. The composition of claim 642 or 643, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 306).
645. The composition of any one of claims 639-644, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 303), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 304), a HC CDR3 sequence of FTVLAMDY (SEQ ID NO.: 305), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 306), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 307), and a LC CDR3 sequence of QQASWLYWVIT (SEQ ID NO.: 308).
646. The composition of any one of claims 639-645, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMD YWGQGTLVTVSS (SEQ ID NO.: 302).
647. The composition of any one of claims 642-646, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWVITFGQGTKV EIK (SEQ ID NO.: 301).
648. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FHDLAMDY (SEQ ID NO.: 313).
649. The composition of claim 648, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 312).
650. The composition of claim 648 or 649, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 311).
651. The composition of any one of claims 648-650, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWKIT (SEQ ID NO.: 316).
652. The composition of claim 651, wherein the VL comprises a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 315).
653. The composition of claim 651 or 652, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 314).
654. The composition of any one of claims 648-653, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 311), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 312), a HC CDR3 sequence of FHDLAMDY (SEQ ID NO.: 313), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 314), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 315), and a LC CDR3 sequence of QQASWLYWKIT (SEQ ID NO.: 316).
655. The composition of any one of claims 648-654, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHDLAMD YWGQGTLVTVSS (SEQ ID NO.: 310).
656. The composition of any one of claims 651-655, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKITFGQGTKV EIK (SEQ ID NO.: 309).
657. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FHELAMDY (SEQ ID NO.: 321).
658. The composition of claim 657, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 320).
659. The composition of claim 657 or 658, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 319).
660. The composition of any one of claims 657-659, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWLIT (SEQ ID NO.: 324).
661. The composition of claim 660, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 323).
662. The composition of claim 660 or 661, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 322).
663. The composition of any one of claims 657-662, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 319), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 320), a HC CDR3 sequence of FHELAMDY (SEQ ID NO.: 321), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 322), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 323), and a LC CDR3 sequence of QQASWLYWLIT (SEQ ID NO.: 324).
664. The composition of any one of claims 657-663, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMD YWGQGTLVTVSS (SEQ ID NO.: 318).
665. The composition of any one of claims 660-664, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKV EIK (SEQ ID NO.: 317).
666. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTVLAMDY (SEQ ID NO.: 329).
667. The composition of claim 666, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 328).
668. The composition of claim 666 or 667, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 327).
669. The composition of any one of claims 666-668, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWKIT (SEQ ID NO.: 332).
670. The composition of claim 669, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 331).
671. The composition of claim 669 or 670, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 330).
672. The composition of any one of claims 666-671, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 327), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 328), a HC CDR3 sequence of FTVLAMDY (SEQ ID NO.: 329), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 330), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 331), and a LC CDR3 sequence of QQSSWLYWKIT (SEQ ID NO.: 332).
673. The composition of any one of claims 666-672, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMD YWGQGTLVTVSS (SEQ ID NO.: 326).
674. The composition of any one of claims 669-673, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKV EIK (SEQ ID NO.: 325).
675. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FELLAMDY (SEQ ID NO.: 337).
676. The composition of claim 675, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 336).
677. The composition of claim 675 or 676, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 335).
678. The composition of any one of claims 675-677, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWKIT (SEQ ID NO.: 340).
679. The composition of claim 678, wherein the VL comprises a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 339).
680. The composition of claim 678 or 679, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 338).
681. The composition of any one of claims 675-680, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 335), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 336), a HC CDR3 sequence of FELLAMDY (SEQ ID NO.: 337), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 338), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 339), and a LC CDR3 sequence of QQSSWLYWKIT (SEQ ID NO.: 340).
682. The composition of any one of claims 675-681, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFELLAMD YWGQGTLVTVSS (SEQ ID NO.: 334).
683. The composition of any one of claims 678-682, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKV EIK (SEQ ID NO.: 333).
684. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTVLAMDY (SEQ ID NO.: 345).
685. The composition of claim 684, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 344).
686. The composition of claim 684 or 685, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 343).
687. The composition of any one of claims 684-686, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWKVT (SEQ ID NO.: 348).
688. The composition of claim 687, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 347).
689. The composition of claim 687 or 688, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 346).
690. The composition of any one of claims 684-689, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 343), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 344), a HC CDR3 sequence of FTVLAMDY (SEQ ID NO.: 345), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 346), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 347), and a LC CDR3 sequence of QQSSWLYWKVT (SEQ ID NO.: 348).
691. The composition of any one of claims 684-690, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMD YWGQGTLVTVSS (SEQ ID NO.: 342).
692. The composition of any one of claims 687-691, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKVTFGQGTKV EIK (SEQ ID NO.: 341).
693. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FYDLAMDY (SEQ ID NO.: 353).
694. The composition of claim 693, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 352).
695. The composition of claim 693 or 694, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 351).
696. The composition of any one of claims 693-695, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWKVT (SEQ ID NO.: 356).
697. The composition of claim 696, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 355).
698. The composition of claim 696 or 697, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 354).
699. The composition of any one of claims 693-698, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 351), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 352), a HC CDR3 sequence of FYDLAMDY (SEQ ID NO.: 353), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 354), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 355), and a LC CDR3 sequence of QQSSWLYWKVT (SEQ ID NO.: 356).
700. The composition of any one of claims 693-699, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMD YWGQGTLVTVSS (SEQ ID NO.: 350).
701. The composition of any one of claims 696-700, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKVTFGQGTKV EIK (SEQ ID NO.: 349).
702. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FHELAMDY (SEQ ID NO.: 361).
703. The composition of claim 702, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 360).
704. The composition of claim 702 or 703, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 359).
705. The composition of any one of claims 702-704, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWLIT (SEQ ID NO.: 364).
706. The composition of claim 705, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 363).
707. The composition of claim 705 or 706, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 362).
708. The composition of any one of claims 702-707, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 359), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 360), a HC CDR3 sequence of FHELAMDY (SEQ ID NO : 361), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 362), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 363), and a LC CDR3 sequence of QQASWLYWLIT (SEQ ID NO.: 364).
709. The composition of any one of claims 702-708, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMD YWGQGTLVTVSS (SEQ ID NO.: 358).
710. The composition of any one of claims 705-709, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKV EIK (SEQ ID NO.: 357).
711. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTVLAMDY (SEQ ID NO.: 369).
712. The composition of claim 711, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 368).
713. The composition of claim 711 or 712, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 367).
714. The composition of any one of claims 711-713, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWKVT (SEQ ID NO.: 372).
715. The composition of claim 714, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 371).
716. The composition of claim 714 or 715, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 370).
717. The composition of any one of claims 711-716, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 367), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 368), a HC CDR3 sequence of FTVLAMDY (SEQ ID NO.: 369), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 370), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 371), and a LC CDR3 sequence of QQASWLYWKVT (SEQ ID NO.: 372).
718. The composition of any one of claims 711-717, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTVLAMD YWGQGTLVTVSS (SEQ ID NO.: 366).
719. The composition of any one of claims 714-718, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWKVTFGQGTKV EIK (SEQ ID NO.: 365).
720. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FYDLAMDY (SEQ ID NO.: 377).
721. The composition of claim 720, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 376).
722. The composition of claim 720 or 721, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 375).
723. The composition of any one of claims 720-722, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWVIT (SEQ ID NO.: 380).
724. The composition of claim 723, wherein the VL comprises a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 379).
725. The composition of claim 723 or 724, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 378).
726. The composition of any one of claims 720-725, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 375), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 376), a HC CDR3 sequence of FYDLAMDY (SEQ ID NO.: 377), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 378), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 379), and a LC CDR3 sequence of QQSSWLYWVIT (SEQ ID NO.: 380).
727. The composition of any one of claims 720-726, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFYDLAMD YWGQGTLVTVSS (SEQ ID NO.: 374).
728. The composition of any one of claims 723-727, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWVITFGQGTKV EIK (SEQ ID NO.: 373).
729. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FHELAMDY (SEQ ID NO.: 385).
730. The composition of claim 729, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 384).
731. The composition of claim 729 or 730, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 383).
732. The composition of any one of claims 729-731, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWLIT (SEQ ID NO.: 388).
733. The composition of claim 732, wherein the VL comprises a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 387).
734. The composition of claim 732 or 733, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 386).
735. The composition of any one of claims 729-734, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 383), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 384), a HC CDR3 sequence of FHELAMDY (SEQ ID NO.: 385), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 386), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 387), and a LC CDR3 sequence of QQASWLYWLIT (SEQ ID NO.: 388).
736. The composition of any one of claims 729-735, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHELAMD YWGQGTLVTVSS (SEQ ID NO.: 382).
737. The composition of any one of claims 732-736, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWLITFGQGTKV EIK (SEQ ID NO.: 381).
738. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FAHLAMDY (SEQ ID NO.: 393).
739. The composition of claim 738, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 392).
740. The composition of claim 738 or 739, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 391).
741. The composition of any one of claims 738-740, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWLIT (SEQ ID NO.: 396).
742. The composition of claim 741, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 395).
743. The composition of claim 741 or 742, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 394).
744. The composition of any one of claims 738-743, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 391), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 392), a HC CDR3 sequence of FAHLAMDY (SEQ ID NO.: 393), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 394), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 395), and a LC CDR3 sequence of QQSSWLYWLIT (SEQ ID NO.: 396).
745. The composition of any one of claims 738-744, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFAHLAMD YWGQGTLVTVSS (SEQ ID NO.: 390).
746. The composition of any one of claims 741-745, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWLITFGQGTKVE IK (SEQ ID NO.: 389).
747. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FEDLAMDY (SEQ ID NO.: 401).
748. The composition of claim 747, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 400).
749. The composition of claim 747 or 748, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 399).
750. The composition of any one of claims 747-749, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLYWYIT (SEQ ID NO.: 404).
751. The composition of claim 750, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 403).
752. The composition of claim 750 or 751, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 402).
753. The composition of any one of claims 747-752, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 399), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 400), a HC CDR3 sequence of FEDLAMDY (SEQ ID NO.: 401), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 402), a LC CDR2 sequence of SASSLYS (SEQ ID NO: 403), and a LC CDR3 sequence of QQASWLYWYIT (SEQ ID NO.: 404).
754. The composition of any one of claims 747-753, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFEDLAMD YWGQGTLVTVSS (SEQ ID NO.: 398).
755. The composition of any one of claims 750-754, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLYWYITFGQGTKV EIK (SEQ ID NO.: 397).
756. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FRTLAMDY (SEQ ID NO.: 409).
757. The composition of claim 756, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 408).
758. The composition of claim 756 or 757, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 407).
759. The composition of any one of claims 756-758, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWIIT (SEQ ID NO.: 412).
760. The composition of claim 759, wherein the VL comprises a LC CDR2 sequence of SASSLYS (SEQ ID NO: 411).
761. The composition of claim 759 or 760, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 410).
762. The composition of any one of claims 756-761, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 407), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 408), a HC CDR3 sequence of FRTLAMDY (SEQ ID NO.: 409), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 410), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 411), and a LC CDR3 sequence of QQSSWLYWIIT (SEQ ID NO.: 412).
763. The composition of any one of claims 756-762, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFRTLAMD YWGQGTLVTVSS (SEQ ID NO.: 406).
764. The composition of any one of claims 759-763, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWIITFGQGTKVEI K (SEQ ID NO.: 405).
765. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FTHYAMDY (SEQ ID NO.: 417).
766. The composition of claim 765, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 416).
767. The composition of claim 765 or 766, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 415).
768. The composition of any one of claims 765-767, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQSSWLYWKIT (SEQ ID NO.: 420).
769. The composition of claim 768, wherein the VL comprises a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 419).
770. The composition of claim 768 or 769, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 418).
771. The composition of any one of claims 765-770, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 415), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 416), a HC CDR3 sequence of FTHYAMDY (SEQ ID NO.: 417), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 418), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 419), and a LC CDR3 sequence of QQSSWLYWKIT (SEQ ID NO.: 420).
772. The composition of any one of claims 765-771, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFTHYAMD YWGQGTLVTVSS (SEQ ID NO.: 414).
773. The composition of any one of claims 768-772, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQSSWLYWKITFGQGTKV EIK (SEQ ID NO.: 413).
774. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FRLYAMDY (SEQ ID NO.: 425).
775. The composition of claim 774, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 424).
776. The composition of claim 774 or 775, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 423).
777. The composition of any one of claims 774-776, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLHWKLT (SEQ ID NO.: 428).
778. The composition of claim 777, wherein the VL comprises a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 427).
779. The composition of claim 777 or 778, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 426).
780. The composition of any one of claims 774-779, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 423), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 424), a HC CDR3 sequence of FRLYAMDY (SEQ ID NO.: 425), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 426), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 427), and a LC CDR3 sequence of QQASWLHWKLT (SEQ ID NO.: 428).
781. The composition of any one of claims 774-780, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFRLYAMD YWGQGTLVTVSS (SEQ ID NO.: 422).
782. The composition of any one of claims 777-781, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLHWKLTFGQGTKV EIK (SEQ ID NO.: 421).
783. A composition comprising a polypeptide comprising an antigen-binding domain, wherein the antigen-binding domain comprises a heavy chain variable region (VH) that comprises a heavy chain complementarity determining region 3 FHRLAMDY (SEQ ID NO.: 433).
784. The composition of claim 774, wherein the VH comprises a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO.: 432).
785. The composition of claim 774 or 775, wherein the VH comprises a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 431).
786. The composition of any one of claims 774-776, wherein the antigen-binding domain comprises a light chain variable region (VL) that comprises a light chain complementarity determining region 3 (LC CDR3) sequence of QQASWLHWKLT (SEQ ID NO.: 436).
787. The composition of claim 777, wherein the VL comprises a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 435).
788. The composition of claim 777 or 778, wherein the VL comprises a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 434).
789. The composition of any one of claims 774-779, wherein the antigen-binding domain comprises: a HC CDR1 sequence of FSSSSIH (SEQ ID NO: 431), a HC CDR2 sequence of SISSSSGSTSYADSVKG (SEQ ID NO: 432), a HC CDR3 sequence of FHRLAMDY (SEQ ID NO.: 433), a LC CDR1 sequence of RASQSVSSAVA (SEQ ID NO: 434), a LC CDR2 sequence of SAS SLYS (SEQ ID NO: 435), and a LC CDR3 sequence of QQASWLHWKLT (SEQ ID NO.: 436).
790. The composition of any one of claims 774-780, wherein the VH comprises a sequence with at least 80% sequence identity to the sequence
EVQLVESGGGL VQPGGSLRLSC AASGFTFS S S SMWVRQAPGKGLEWVASIS S SSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARFHRLAMD YWGQGTLVTVSS (SEQ ID NO.: 430).
791. The composition of any one of claims 777-781, wherein the VL comprises a sequence with at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASS LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASWLHWKLTFGQGTKV EIK (SEQ ID NO.: 429).
792. A pharmaceutical composition comprising a polypeptide of any one of claims 595- 791, and a pharmaceutically acceptable carrier, excipient, adjuvant or diluent.
793. A method of treating cancer in a subject in need thereof wherein the subject has been treated with a free targeted covalent inhibitor, the method comprising administering to the subject the polypeptide of any one of claims 595-791 or the pharmaceutical composition of claim 792.
794. The method of claim 793, wherein the free targeted covalent inhibitor is sotorasib, adagrasib, or divarasib.
795. The method of claim 793 or 794, wherein the subject is refractory to a treatment with the free targeted covalent inhibitor.
796. A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject the polypeptide of any one of claims 595-791 or the pharmaceutical composition of claim 792.
797. The method of claim 796, wherein the method further comprises administering simultaneously a small molecule drug.
798. A method of administering to the subject in need thereof the polypeptide of any one of claims 595-791 or the pharmaceutical composition of claim 792, and a free targeted covalent inhibitor, wherein the free targeted covalent inhibitor is optionally sotorasib, adagrasib, or divarasib.
799. The method of claim 798, wherein the polypeptide of any one of claims 595-791 or the pharmaceutical composition of claim 792 is administered after administration of the free targeted covalent inhibitor or simultaneously with the free targeted covalent inhibitor.
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