Compositions and methods comprising antibodies that bind to covalent peptide conjugates

Abstract

Compositions and methods are provided that include a binding partner that specifically binds to a peptide conjugate / MHC complex comprising a peptide conjugate formed by a covalent reaction of a targeted covalent inhibitor with a peptide. The binding partners are provided as antibodies and antibody derivatives that specifically bind to the peptide conjugate / MHC complex.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 366,819, filed June 22, 2022, U.S. Provisional Application No. 63 / 369,702, filed July 28, 2022, U.S. Provisional Application No. 63 / 402,606, filed August 31, 2022, U.S. Provisional Application No. 63 / 377,466, filed September 28, 2022, and U.S. Provisional Application No. 63 / 485,788, filed February 17, 2023, each of which is incorporated herein by reference in its entirety.

[0003] Statement Regarding Federally Funded Research

[0004] This invention was made with government support under Grant Nos. CA016087 and CA267362 awarded by the National Institutes of Health. The government has certain rights in this invention. Background Art

[0005] There is a continuing and unmet need for agents that can bind to targets, including drugs that covalently bind to proteins or peptides. In particular, there is a need to improve the efficacy of targeted therapies while increasing the immunogenicity of tumors and the efficacy of immunotherapies for cancers caused by the loss of intracellular oncogenes and tumor suppressor genes. The present disclosure is relevant to these needs. Summary of the Invention

[0006] The present disclosure provides compositions and methods comprising binding partners that specifically bind to a target site on a protein or peptide comprising a covalently linked molecule. This is believed to be the first disclosure of a binding partner with such binding functionality. The present disclosure illustrates this method using binding partners in the form of various antibodies and antibody derivatives that specifically bind to proteins and peptides that have been covalently modified by linking molecules, wherein the molecules are exemplified by various drugs. In addition, the present disclosure also demonstrates that binding partners that specifically bind to peptides that have been covalently modified by linking small molecule drugs have specificity for the covalently modified peptide when presented in the context of human leukocyte antigen (HLA), where HLA is a representative example of the major histocompatibility complex (MHC). Thus, a binding partner with specificity for a peptide-drug conjugate in an HLA complex is demonstrated. The present disclosure includes polynucleotides encoding the binding partner and cells modified to express the binding partner. The present disclosure includes diagnostic, prophylactic, and therapeutic methods using the binding partner.

[0007] In one 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 a covalent reaction of a targeted covalent inhibitor or fragment thereof with a peptide; and (b) MHC.

[0008] In some embodiments, the binding affinity of the binding partner to the peptide conjugate / MHC complex is greater than the binding affinity to the peptide or free targeted covalent inhibitor. In some embodiments, the affinity of the binding partner to the peptide conjugate / MHC complex is 100-10,000 times greater than the affinity of the binding partner to the peptide or free targeted covalent inhibitor.

[0009] In some embodiments, the MHC is a human leukocyte antigen (HLA), optionally wherein the HLA is HLA-A, HLA-B, or HLA-C. In some embodiments, the HLA molecule is an HLA-A*02:01, HLA-A*03:01, HLA-A*01:01, HLA-A*11:01, HLA-A*24:02, HLA-A*26:01, HLA-B*07:02, HLA-B*08:01, HLA-B*27:05, HLA-B*39:01, HLA-B*40:01, HLA-B*58:01, and / or HLA-B*15:01 molecule.

[0010] In some embodiments, the peptide comprises a nucleophilic or electrophilic residue, which is optionally one of cysteine, lysine, tyrosine, histidine, serine, arginine, or threonine.

[0011] In some embodiments, the peptide comprises a cysteine ​​residue.

[0012] In some embodiments, the peptide conjugate is formed by a covalent reaction between a targeted covalent inhibitor and a cysteine ​​residue in the peptide.

[0013] In some embodiments, the peptide is a fragment of a protein associated with cancer, optionally where the protein is encoded by a gene that is mutated in the cancer.

[0014] In some embodiments, the peptide is a fragment 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.

[0015] In some embodiments, the peptide is or is derived from RAS (e.g., KRAS, HRAS, or NRAS), Bruton's tyrosine kinase (BTK), any epidermal growth factor receptor (EGFR) family member selected from EGFR (ERBB1), HER2 / NEU (ERBB2), HER3 (ERBB3), and HER4 (ERBB4); MET (HGFR); any fibroblast growth factor receptor (FGFR); any cyclin-dependent kinase (CDK); acetylcholinesterase (ACHE); p90 ribosomal S 6 kinase (RSK); TP53, IDH1, GNAS, FBXW7, CTNNB1, DNMT3A, a cathepsin selected from cathepsins B, C, F, H, K, L, O, S, V, W and X; any caspase; a protein involved in obesity, which is optionally pancreatic lipase or METAP2; any cancer testis antigen (CTA); long interspersed element-1 (LINE-1); short interspersed elements, which are optionally Alu; and any endogenous retroviral protein, and optionally wherein RAS is KRAS, HRAS or NRAS.

[0016] In some embodiments, the peptide comprises KRAS G12C , KRAS G12D 、KRAS G12R , KRAS G12S , HRAS G12C , HRAS G12D , HRAS G12R , HRAS G12S NRAS G12C NRAS G12D NRAS G12R or NRAS G12S fragment.

[0017] In some embodiments, the peptide comprises the amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV.

[0018] In some embodiments, the targeted covalent inhibitor is (i) a ternary complex KRAS G12C Inhibitors or KRAS G12C Degrader, (ii) Ternary complex KRAS G12D Inhibitors or KRAS G12D Degrader, (iii) Ternary complex KRAS G12R Inhibitors or KRAS G12R Degraders, or (iv) ternary complex KRAS G12S Inhibitors or KRAS G12S Degradants.

[0019] In some embodiments, the peptide comprises KRAS G12C mutations, and targeted covalent inhibitors are KRAS G12C In some embodiments, the peptide comprises a KRAS G12D mutations, and targeted covalent inhibitors are KRAS G12D In some embodiments, the peptide comprises a KRAS G12R mutations, and targeted covalent inhibitors are KRAS G12R In some embodiments, the peptide comprises a KRAS G12S mutations, and targeted covalent inhibitors are KRAS G12S Inhibitor.

[0020] In some embodiments, the targeted covalent inhibitor is osimertinib, ibrutinib, neratinib, sotorasib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1, or BI 182391.

[0021] In some embodiments, the peptide comprises a fragment of EGFR.

[0022] In some embodiments, the peptide comprises the amino acid sequence of QLMPFGCLL, LMPFGCLLDY, or MPFGCLLDY.

[0023] In some embodiments, the peptide comprises a fragment of EGFR and the targeted covalent inhibitor is an inhibitor of an EGFR family kinase.

[0024] In some embodiments, the targeted covalent inhibitor is osimertinib or neratinib.

[0025] In some embodiments, the peptide comprises a fragment of BTK.

[0026] In some embodiments, the peptide comprises the amino acid sequence of YMANGCLLNY.

[0027] In some embodiments, the targeted covalent inhibitor is ibrutinib.

[0028] In some embodiments, the peptide is a full-length protein that is processed in the cell following covalent reaction with the targeted covalent inhibitor to produce smaller peptide fragments.

[0029] In some embodiments, the peptide conjugate comprises a compound selected from the group consisting of Compounds 1-8:

[0030]

[0031]

[0032]

[0033] It is covalently bound to a cysteine ​​residue in a peptide comprising the amino acid sequence of VVVGACGVGK, VVGACGVGK or KLVVVGACGV.

[0034] In some embodiments, the MHC is HLA-A*02:01, HLA-A*03:01, and / or HLA-A*11:01.

[0035] 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.

[0036] In some embodiments, the peptide conjugate comprises Compound 9:

[0037]

[0038] It is covalently bound to a cysteine ​​residue in a peptide comprising the amino acid sequence of QLMPFGCLL, LMPFGCLLDY or MPFGCLLDY.

[0039] In some embodiments, the MHC is HLA-A*02, HLA-A*01, HLA-A*03, or HLA-A*26.

[0040] In some embodiments, the peptide conjugate comprises Compound 10:

[0041]

[0042] It is covalently bound to a cysteine ​​residue in a peptide comprising the amino acid sequence of YMANGCLLNY.

[0043] In some embodiments, the MHC is HLA-A*01:01.

[0044] In some embodiments, the binding partner specifically binds to a peptide conjugate / MHC complex, wherein the peptide conjugate is composed of sotolacib and KRAS G12CIn some embodiments, 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 CDR-H1 comprising the amino acid sequence of DYSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a CDR-H2 comprising the amino acid sequence of SISSSSGSTSYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and / or (iii) a CDR-H3 comprising the amino acid sequence of GX1WX2X3AMDY, wherein X1 is G, R, H, S, or K, X2 is Y or I, and X3 is P or A. In some embodiments, VL comprises: (i) a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a CDR-L2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and / or (iii) a CDR-L3 comprising the amino acid sequence of QQX1SYVX2X3X4IT, wherein X1 is I, A, P, V, or S, X2 is K, R, A, or H, X3 is K or R, and X4 is L, T, K, R, V, A, or E.

[0045] In some embodiments, the binding partner specifically binds to a peptide conjugate / MHC complex, wherein the peptide conjugate is composed of sotolacib and KRAS G12C The invention relates to a method for forming an oligonucleotide comprising: covalently reacting a peptide to form an oligonucleotide, wherein the binding partner comprises the CDR-H1, CDR-H2 and CDR-H3 amino acid sequences of the VH amino acid sequence and / or the CDR-L1, CDR-L2 and CDR-L3 amino acid sequences of the VL amino acid sequence of the binding partner selected from the group consisting of: RA_D11, RA_D01-RA_D04, RA_D06-RA_D09, RA_D12-RA_D14, RA_D16, RA_D18-RA_D21, RA_D23 and RA_D24; or a variant thereof comprising 1-5 amino acid changes in one or more CDR amino acid sequences.

[0046] In some embodiments, the binding partner comprises a CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and / or CDR-L3 amino acid sequence of a binding partner selected from the group consisting of RA_D11, RA_D01-RA_D04, RA_D06-RA_D09, RA_D12-RA_D14, RA_D16, RA_D18-RA_D21, RA_D23 and RA_D24 (Table G and Table H), or a variant thereof comprising 1-5 amino acid changes in one or more CDR amino acid sequences.

[0047] In some embodiments, the binding partner comprises a VH and / or VL amino acid sequence that is 90%, 95%, or 100% identical to the following VH and VL sequences:

[0048]

[0049]

[0050]

[0051] 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 osimertinib with an EGFR peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL). In some embodiments, VH comprises: (i) a CDR-H1 comprising the amino acid sequence of SSYIH, or a variant thereof comprising 1-5 amino acid changes; (ii) a CDR-H2 comprising the amino acid sequence of YISPSYGSTSYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and / or (iii) a CDR-H3 comprising the amino acid sequence of EX1X2X3MX4X5DY, wherein X1 is Y, L, S, or E, X2 is V, T, or I, X3 is T or I, X4 is A, T, or S, and X5 is L, A, I, K, P, or T. In some embodiments, VL comprises: (i) a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a CDR-L2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes; and / or (iii) a CDR-L3 comprising the amino acid sequence of QQYX1X2WPX3T, wherein X1 is S or A, or S; X2 is Y, H, A, D, E, K, S or G; and X3 is I or E.

[0052] In some embodiments, the binding partner specifically binds to the peptide conjugate / MHC complex, wherein the peptide conjugate is formed by the covalent reaction of osimertinib and the EGFR peptide, wherein the binding partner comprises the CDR-H1, CDR-H2, and CDR-H3 amino acid sequences of the VH amino acid sequence and / or the CDR-L1, CDR-L2, and CDR-L3 amino acid sequences of the VL amino acid sequence of the binding partner selected from the group consisting of: OEA2-5, EO_Q01-EO_Q18, and EO_Q20-EO_Q24, or variants thereof comprising 1-5 amino acid changes in one or more CDR amino acid sequences.

[0053] In some embodiments, the binding partner comprises a CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and / or CDR-L3 amino acid sequence of a binding partner selected from the group consisting of: OEA2-5, EO_Q01-EO_Q18 and EO_Q20-EO_Q24 (Tables I and J), or a variant thereof comprising 1-5 amino acid changes in one or more CDR amino acid sequences.

[0054] In some embodiments, the binding partner comprises a VH and / or VL amino acid sequence that is 90%, 95%, or 100% identical to the following VH and VL sequences:

[0055]

[0056]

[0057]

[0058] In some embodiments, the binding partner has a higher affinity for a peptide conjugate / MHC complex comprising a first HLA than for a peptide conjugate / MHC complex comprising a second HLA.

[0059] In some embodiments, the first HLA molecule and the second HLA molecule are each selected from the group consisting of HLA-A*02:01, HLA-A*03:01, HLA-A*01:01, HLA-A*11:01, HLA-A*24:02, HLA-A*26:01, HLA-B*07:02, HLA-B*08:01, HLA-B*27:05, HLA-B*39:01, HLA-B*40:01, HLA-B*58:01 and / or HLA-B*15:01.

[0060] In some embodiments, the binding partner is a complete antibody, a bispecific antibody, a multispecific antibody, an antigen-binding (Fab) fragment, a Fab' fragment, a (Fab')2 fragment, a Fd, a Fv, a dAb, a single domain fragment or a single monomeric variable antibody domain, a dual affinity retargeting (DART) molecule, a diabody (Db), a single chain diabody (scDb), a single chain variable fragment (scFv), a bispecific T cell engaging antibody (engager) (BiTE), a bispecific killer cell engaging antibody (BiKE), a CrossMab, a camelid antibody, a trispecific binding partner, a chimeric antigen receptor (CAR), a monomer (Monobody) (also known as Adnectin), a DARPin, an anticarrier protein (anticalin), an affibody (affibody) or an affimer. In some embodiments, the binding partner is bispecific. In some embodiments, the binding partner specifically binds to a peptide conjugate / MHC complex and a T cell antigen. In some embodiments, the binding partner specifically binds to a peptide conjugate / MHC complex and human CD3.

[0061] In some embodiments, the binding partner comprises the VH and / or VL amino acid sequence of UCHT1:

[0062]

[0063] In some embodiments, the binding partner comprises a sequence that is at least 90% similar to any of the following sequences excluding the underlined sequences:

[0064]

[0065]

[0066] In various embodiments, the binding partners described herein do not comprise the amino acid sequence LEGGGGLNDIFEAQKIEWHESRHHHHHH. In various embodiments, the binding partners described herein do not comprise an AviTag or His tag. In various embodiments, the binding partner to be administered to a subject does not comprise the amino acid sequence LEGGGGLNDIFEAQKIEWHESRHHHHHH. In various embodiments, the binding partner to be administered to a subject does not comprise an AviTag or His tag.

[0067] In some embodiments, the binding partner is a single-chain diabody (scDb) and comprises a sequence at least 90% identical to any of the following sequences excluding the underlined sequences:

[0068]

[0069]

[0070] In some embodiments, the binding partner comprises a heavy chain constant region selected from the group consisting of human IgM, IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2.

[0071] In some embodiments, the heavy chain constant region comprises one or more amino acid substitutions in the Fc region.

[0072] In some embodiments, the binding partner comprises a human kappa light chain constant region or a human lambda light chain constant region.

[0073] In some embodiments, the binding partner is in the form of a CrossMab.

[0074] In some embodiments, the binding partner is conjugated to a detectable label, chemotherapeutic agent, radioisotope, or toxin.

[0075] In some embodiments, the binding partner is contained within a chimeric antigen receptor.

[0076] In some embodiments, the binding partner is expressed by T cells, macrophages, neutrophils, or natural killer cells.

[0077] In some embodiments, binding of the binding partner to the peptide conjugate / MHC complex is not inhibited by a free targeted covalent inhibitor.

[0078] In another aspect, the present disclosure provides a complex comprising a binding partner disclosed herein and a peptide conjugate / MHC complex.

[0079] In another aspect, the present disclosure provides polynucleotides encoding the binding partners disclosed herein.

[0080] In another aspect, the disclosure provides polynucleotides encoding the heavy chain variable region and / or light chain variable region of a binding partner disclosed herein.

[0081] In another aspect, the present disclosure provides a vector comprising a polynucleotide disclosed herein.

[0082] In some embodiments, the vector is a viral vector. In some embodiments, the viral vector is an adenoviral vector, a lentiviral vector, a retroviral vector, or an adeno-associated viral vector.

[0083] In another aspect, the present disclosure provides a recombinant host cell comprising (a) a polynucleotide disclosed herein; (b) a vector disclosed herein; (c) a first polynucleotide encoding the VH or heavy chain of a binding partner disclosed herein, and a second polynucleotide encoding the VL or light chain of a binding partner disclosed herein; or (d) a first vector comprising a first polynucleotide encoding the VH or heavy chain of a binding partner disclosed herein, and a second vector comprising a second polynucleotide encoding the VL or light chain of a binding partner disclosed herein.

[0084] In another aspect, the present disclosure provides a pharmaceutical composition comprising a binding partner disclosed herein, a polynucleotide disclosed herein, a vector disclosed herein, or a host cell disclosed herein, and a pharmaceutically acceptable carrier or excipient.

[0085] In another aspect, the present disclosure provides a method of producing a binding partner, the method comprising culturing a host cell disclosed herein under suitable conditions such that the polynucleotide is expressed and the binding partner is produced.

[0086] In another aspect, the present disclosure provides a eukaryotic cell comprising a polynucleotide disclosed herein or a 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 γδ T cell, optionally a natural killer cell, a natural killer T cell, a mucosal-associated invariant T (MAIT) cell, a neutrophil or a macrophage.

[0087] In another aspect, the present disclosure provides a method comprising administering a binding partner disclosed herein, a polynucleotide disclosed herein, a vector disclosed herein, a pharmaceutical composition disclosed herein, or a cell disclosed herein to an individual in need thereof.

[0088] In another aspect, the present disclosure provides a method for producing 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.

[0089] 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) MHC.

[0090] In some embodiments, the peptide comprises a fragment of RAS (e.g., KRAS G12C 、KRAS G12D 、KRASG12R 、KRAS G12S , HRAS G12C , HRAS G12D , HRAS G12R , HRAS G12S NRAS G12C NRAS G12D NRAS G12R or NRAS G12S ), EGFR, BTK, HER2 / NEU (ERBB2), HER3 (ERBB3), HER4 (ERBB4), MET (HGFR); FGFR, CDK, acetylcholinesterase (ACHE), p90 ribosomal S6 kinase (RSK), TP53, IDH1, GNAS, FBXW7, CTNNB1, DNMT3A, cathepsin B, cathepsin C, cathepsin F, cathepsin H, cathepsin K, cathepsin L, cathepsin O, cathepsin S, cathepsin V, cathepsin W, cathepsin X, caspase, pancreatic lipase, METAP2, any cancer testis antigen (CTA), long interspersed element-1 (LINE-1), short interspersed elements that are optionally Alu, or any endogenous retroviral protein.

[0091] In some embodiments, the targeted covalent inhibitor is osimertinib, ibrutinib, neratinib, sotolacib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1, or BI 182391. 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 4-8,

[0092]

[0093]

[0094] It covalently binds to a cysteine ​​residue in a peptide comprising the amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV; and (b) MHC.

[0095] In another aspect, the present disclosure provides a cell-free peptide conjugate / MHC complex comprising (a) Compound 2:

[0096]

[0097] It covalently binds to a cysteine ​​residue in a peptide comprising the amino acid sequence of QLMPFGCLL, LMPFGCLLDY or MFPGCLLDY; and (b) MHC.

[0098] In another aspect, the present disclosure provides a cell-free peptide conjugate / MHC complex comprising (a) Compound 3:

[0099]

[0100] It covalently binds to a cysteine ​​residue in a peptide comprising the amino acid sequence of YMANGCLLNY; and (b) MHC.

[0101] In some embodiments, the MHC is an HLA, optionally wherein the HLA is an HLA-A*02:01, HLA-A*03:01, HLA-A*01:01, HLA-A*11:01, HLA-A*24:02, HLA-A*26:01, HLA-B*07:02, HLA-B*08:01, HLA-B*27:05, HLA-B*39:01, HLA-B*40:01, HLA-B*58:01, or an HLA-B*15:01 molecule.

[0102] In another aspect, the present disclosure provides a recombinant cell or particle comprising on its outer surface a cell-free peptide conjugate / MHC complex disclosed herein.

[0103] In another aspect, the present disclosure provides a method for identifying one or more binding partners that specifically bind to a peptide conjugate presented by HLA, the method comprising contacting a cell-free peptide conjugate / MHC complex disclosed herein with a plurality of binding partners, and selecting one or more binding partners that specifically bind to the cell-free peptide conjugate / MHC complex. In some embodiments, the method further comprises determining the sequence of the selected one or more binding partners. In some embodiments, the method further comprises generating the selected one or more binding partners.

[0104] 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 cell sample from a subject who has been treated with a targeted covalent inhibitor and has different HLA types, or providing a cell-free peptide conjugate / MHC complex disclosed herein presented by two or more HLAs, and screening for binding partners to identify a binding partner that specifically binds to a peptide conjugate or antigen presented by more than one HLA. In some embodiments, the two or more HLA types include HLA-A*02:01 and at least one additional HLA type. In some embodiments, the two or more HLAs are each an HLA-A*02:01, HLA-A*03:01, HLA-A*01:01, HLA-A*11:01, HLA-A*24:02, HLA-A*26:01, HLA-B*07:02, HLA-B*08:01, HLA-B*27:05, HLA-B*39:01, HLA-B*40:01, HLA-B*58:01, or an HLA-B*15:01 molecule.

[0105] In another aspect, the present disclosure provides a method of killing cancer cells in a subject, the method comprising administering to the subject: (a) a targeted covalent inhibitor, and (b) a binding partner disclosed herein, a polynucleotide disclosed herein, a vector disclosed herein, a pharmaceutical composition disclosed herein, or a cell disclosed herein.

[0106] In some embodiments, the targeted covalent inhibitor targets RAS (e.g., KRAS, HRAS, NRAS), Bruton's tyrosine kinase (BTK), EGFR (ERBB1), HER2 / NEU (ERBB2), HER3 (ERBB3), HER4 (ERBB4), fibroblast growth factor receptor (FGFR), MET, BRAF, cyclin-dependent kinase (CDK), acetylcholinesterase (ACHE), TP53, IDH1, GNAS, FBXW7, CTNNB1, DNMT3A, cathepsins, caspases, pancreatic lipase, METAP2, any cancer testis antigen, endogenous retroviral protein, long interspersed element-1 (LINE-1), or short interspersed element (SINE).

[0107] In another aspect, the present disclosure provides a method for targeting an already used EGFR, BTK or RAS (e.g., KRAS G12C 、KRAS G12D 、KRAS G12R 、KRAS G12S , HRAS G12C , HRAS G12D , HRASG12R , HRAS G12S NRAS G12C NRAS G12D NRAS G12R or NRAS G12S ) in subjects who are being treated with targeted covalent inhibitors expressing EGFR, BTK, or RAS (e.g., KRAS G12C 、KRAS G12D 、KRAS G12R 、KRAS G12S , HRAS G12C , HRAS G12D , HRAS G12R , HRAS G12S NRAS G12C NRAS G12D NRAS G12R or NRAS G12S ) comprising administering to a subject a binding partner disclosed herein, a polynucleotide disclosed herein, a vector disclosed herein, a pharmaceutical composition disclosed herein, or a cell disclosed herein.

[0108] In some embodiments, the subject has cancer.

[0109] In another aspect, the present disclosure provides a method of treating cancer in a subject who has been treated with a targeted covalent inhibitor, the method comprising administering to the subject a binding partner disclosed herein, a polynucleotide disclosed herein, a vector disclosed herein, a pharmaceutical composition disclosed herein, or a cell disclosed herein.

[0110] In another aspect, the present disclosure provides a method of treating a disease or disorder in a subject who has been treated with a targeted covalent inhibitor, the method comprising administering to the subject a binding partner disclosed herein, a polynucleotide disclosed herein, a vector disclosed herein, a pharmaceutical composition disclosed herein, or a cell disclosed herein. In some embodiments, the disease or disorder is an autoimmune disease or a fibrotic disease.

[0111] In another aspect, the present disclosure provides a method for enhancing the expression of RAS (e.g., KRAS) in a subject having a cancer exhibiting a RAS or EGFR mutation. G12C 、KRAS G12D 、KRAS G12R 、KRAS G12S , HRAS G12C , HRAS G12D , HRAS G12R , HRAS G12S NRAS G12C NRAS G12D NRASG12R or NRAS G12S ) or EGFR mutations, the method comprising administering to a subject: (a) a RAS (e.g., KRAS G12C 、KRAS G12D 、KRAS G12R 、KRAS G12S , HRAS G12C , HRAS G12D , HRAS G12R , HRAS G12S NRAS G12C NRAS G12D NRAS G12R or NRAS G12S ) or EGFR inhibitors, and binding partners disclosed herein, polynucleotides disclosed herein, vectors disclosed herein, pharmaceutical compositions disclosed herein, or cells disclosed herein. In some embodiments, the subject has previously been treated with RAS (e.g., KRAS G12C 、KRAS G12D 、KRAS G12R 、KRAS G12S , HRAS G12C , HRAS G12D , HRAS G12R , HRAS G12S NRAS G12C NRAS G12D NRAS G12R or NRAS G12S ) or EGFR inhibitor treatment.

[0112] In some embodiments, the inhibitor is osimertinib, ibrutinib, neratinib, sotolacilib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1, or BI182391. In some embodiments, the targeted covalent inhibitor targets RAS (e.g., KRAS, HRAS, or NRAS), Bruton's tyrosine kinase (BTK), EGFR (ERBB1), HER2 / NEU (ERBB2), HER3 (ERBB3), HER4 (ERBB4), fibroblast growth factor receptor (FGFR), MET, BRAF, cyclin-dependent kinase (CDK), acetylcholinesterase (ACHE), TP53, IDH1, GNAS, FBXW7, CTNNB1, DNMT3A, cathepsins, caspases, pancreatic lipase, METAP2, any cancer testis antigen, endogenous retroviral protein, long interspersed element-1 (LINE-1), or short interspersed element (SINE).

[0113] 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 (or colorectal 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.

[0114] In some embodiments, the targeted covalent inhibitor is osimertinib, ibrutinib, neratinib, sotolacilib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1, or BI 182391.

[0115] In some embodiments, the methods disclosed herein further comprise 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-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-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 anchored IL2 or engineered IL2; or an extracellular adenosine (eADO) signaling inhibitor.

[0116] In another aspect, the present disclosure provides a method for 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 a covalent reaction of a targeted covalent inhibitor with a peptide. In some embodiments, the peptide is EGFR, BTK, or RAS (e.g., KRAS G12C , KRAS G12D , KRAS G12R , KRAS G12S , HRAS G12C , HRAS G12D , HRAS G12R , HRAS G12S NRAS G12C NRAS G12D NRAS G12R or NRAS G12S ) peptide. In some embodiments, the targeted covalent inhibitor is EGFR, BTK or RAS (e.g., KRAS G12C , KRAS G12D , KRAS G12R , KRAS G12S , HRAS G12C , HRAS G12D , HRAS G12R , HRAS G12S NRAS G12C NRAS G12D NRAS G12R or NRAS G12S) inhibitor. In some embodiments, the targeted covalent inhibitor is osimertinib, ibrutinib, neratinib, sotolacib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1, or BI 182391. In some embodiments, the biological sample is blood or serum.

[0117] In another aspect, the present disclosure provides a method of identifying a cell comprising a peptide conjugate / MHC complex disclosed herein, the method comprising contacting the cell with a binding partner disclosed herein.

[0118] In another aspect, the present disclosure provides a method for producing one or more binding partners that specifically bind to a peptide conjugate, wherein the peptide conjugate comprises a peptide covalently bound to a non-peptide molecule, the method comprising: (a) exposing the peptide conjugate to a plurality of binding partners, and b) selecting a binding partner that specifically binds to the peptide conjugate to provide the selected one or more binding partners. In some embodiments, the methods disclosed herein further comprise selecting a binding partner that specifically binds to the peptide conjugate but does not detectably bind to or binds to the peptide or the non-peptide molecule with a low affinity when the peptide and the non-peptide molecule are not covalently bound.

[0119] In another aspect, the present disclosure provides a method for producing one or more binding partners that specifically bind to a peptide conjugate presented in the context of an MHC molecule, or a fragment or derivative thereof, wherein the peptide conjugate comprises a peptide covalently bound to a non-peptide molecule, the method comprising: (a) providing a complex of the peptide conjugate and the MHC molecule, or a fragment or derivative thereof; (b) exposing the complex to a plurality of binding partners, and (c) selecting a binding partner that specifically binds to the complex to provide the selected one or more binding partners. In some embodiments, the methods disclosed herein further comprise selecting a binding partner that specifically binds to the complex but does not detectably bind or binds with a lower affinity to a complex of the peptide and the MHC molecule, or a fragment thereof, wherein the peptide is not covalently bound to the non-peptide molecule. In some embodiments, the methods disclosed herein further comprise selecting a binding partner that specifically binds to the peptide conjugate presented in the context of two or more different MHC molecules.

[0120] In some embodiments, the MHC molecule is an MHC class I molecule and the peptide is 7-15 amino acids in length, or wherein the MHC molecule is a non-classical MHC class I molecule and the peptide is 7-15 amino acids in length. In some embodiments, the MHC molecule is an MHC class II molecule and the peptide is 9-30 amino acids in length.

[0121] In some embodiments, the complex is immobilized on a solid support. In some embodiments, the complex is presented on the surface of a cell.

[0122] In some embodiments, the method further comprises determining whether the 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 the cell, or by using the binding partner as an immunotoxin or as an antibody-drug conjugate (ADC) or as a radioconjugate. In some embodiments, the plurality of binding partners is generated from a phage display library or a yeast display library.

[0123] In another aspect, the present disclosure provides a method for killing cancer cells, the method comprising administering to the cells a binding partner that specifically binds to a peptide conjugate, wherein the peptide conjugate comprises (i) a peptide derived from a target protein within the cell, the peptide being covalently bound to (ii) a non-peptide molecule, and wherein the binding partner mediates immune cell-mediated killing of the cells or antibody-drug conjugate (ADC)-mediated, antibody-dependent cellular phagocytosis (ADCP) or complement-dependent cytotoxicity (CDC) immunotoxin killing, or by using the binding partner as an immunotoxin, or by the action of a radioactive conjugate, thereby killing the cancer cells.

[0124] In some embodiments, the peptide conjugate is presented on the cell in the context of an MHC molecule. In some embodiments, the binding partner specifically binds to a complex comprising the peptide conjugate and the MHC molecule. In some embodiments, the binding partner does not detectably bind to a complex of the peptide and the MHC molecule, wherein the peptide is not covalently bound to the non-peptide molecule.

[0125] In some embodiments, the methods disclosed herein further comprise administering the non-peptide molecule to the cell, wherein the non-peptide molecule forms a covalent bond with the target protein in the cell. In some embodiments, the non-peptide molecule is administered to the cell prior to administering the binding partner. In some embodiments, the cell is isolated from a subject. In some embodiments, the cell is in a subject, and the binding partner is administered to the subject. In some embodiments, the cell is in a subject, and the non-peptide molecule and the binding partner are administered to the subject. In some embodiments, the target protein is selectively spliced ​​or overexpressed in cancer cells, but not selectively spliced ​​or overexpressed by cancer cells. In some embodiments, the target protein is encoded by a gene that mutates in cancer cells but does not mutate in non-cancerous cells. In some embodiments, the non-peptide molecule is a covalent inhibitor of the target protein.

[0126] In some embodiments, the binding partner is a complete antibody, a bispecific antibody, a multispecific antibody, an antigen binding (Fab) fragment, a Fab' fragment, a (Fab')2 fragment, a Fd, a Fv, a dAb, a single domain fragment or a single monomer variable antibody domain, a single chain diabody (scDb), a diabody (Db), a dual affinity retargeting (DART) molecule, a single chain variable fragment (scFv), a bispecific T cell engaging antibody (BiTE), a bispecific killer cell engaging antibody (BiKE), a CrossMab, a camelid antibody, a trispecific binding partner, a chimeric antigen receptor (CAR), a monomer (also known as Adnectin), a DARPin, an anticarrier protein, an affibody or an antibody-like protein. In some embodiments, CAR is present on T cells, natural killer (NK) cells, neutrophils or macrophages. In some embodiments, the binding partner is an antibody-drug conjugate (ADC), a radioconjugate or a toxin conjugate.

[0127] In some embodiments, the target protein is selected from RAS (e.g., KRAS, HRAS, or NRAS), Bruton's tyrosine kinase (BTK), epidermal growth factor receptor (EGFR) family members, fibroblast growth factor receptor (FGFR), MET, BRAF, cyclin-dependent kinase (CDK), acetylcholinesterase (ACHE), TP53, IDH1, GNAS, FBXW7, CTNNB1, DNMT3A, cathepsin, caspase, pancreatic lipase, METAP2, cancer testis antigen, viral polymerase, protein required for viral cell entry, protein encoded by a transposable element (e.g., endogenous retrovirus), or a mutant thereof. In some embodiments, the target protein is a KRAS protein comprising a G12C mutation, and the non-peptide molecule is selected from sotolacib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, RMC-6291, DC-6036, D-1553, 2E07, 6H05, SML-8-73-1 or BI 182391 and derivatives thereof. In some embodiments, the target protein is EGFR, and the non-peptide molecule is selected from PD168393, PF00299804 (dacomitinib), EKB569 (pelitinib), afatinib, WZ4002, osimertinib (AZD9291), PF-06459988, nazatinib, naquintinib, omotinib, avitinib, rociterinib, neratinib, pyrotinib, poziotinib and its derivatives. In some embodiments, the target protein is Bruton's tyrosine kinase (BTK), and the non-peptide molecule is selected from ibrutinib, acalabrutinib, zanubrutinib, CHMFL-BTK-11, ONO / GS-405, PRN1008, CC-292 and its derivatives. In some embodiments, the target protein is p90 ribosomal S6 kinase (RSK), and the non-peptide molecule is fluoromethyl ketone (FMK), dimethyl fumarate or its derivatives. In some embodiments, the target protein is FGFR, and the non-peptide molecule is selected from FIIN-1, FIIN-2, FIIN-3, BGJ398, AZD4547, PRN1371, FGF401, and derivatives thereof.

[0128] In another aspect, the present disclosure provides a method of treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of a binding partner that specifically binds to a peptide conjugate, wherein the peptide conjugate comprises (i) a peptide derived from a target protein present in a cancer cell of the subject, the peptide being covalently bound to (ii) a non-peptide molecule, and wherein the binding partner mediates immune cell-mediated killing of the cancer cell or antibody-drug conjugate (ADC)-mediated killing of the cancer cell, antibody-dependent cellular phagocytosis (ADCP) or complement-dependent cytotoxicity (CDC)-mediated killing of the cancer cell, or by using the binding partner as an immunotoxin or radioconjugate in the subject, thereby killing the cancer cell. In some embodiments, the peptide conjugate is presented on the cancer cell of the subject in the context of an MHC molecule. In some embodiments, the binding partner specifically binds to a complex comprising the peptide conjugate and the MHC molecule. In some embodiments, the binding partner does not detectably bind to a complex of the peptide and the MHC molecule, wherein the peptide is not covalently bound to the non-peptide molecule. In some embodiments, the subject has previously received the non-peptide molecule. In some embodiments, the methods disclosed herein further comprise administering to the subject an effective amount of the non-peptide molecule, wherein the non-peptide molecule forms a covalent bond with the target protein in the subject's cancer cells. In some embodiments, the non-peptide molecule is administered to the subject prior to administering the binding partner.

[0129] In another aspect, the present disclosure provides a method for improving the efficacy of an anti-cancer therapy in a subject in need thereof, wherein the anti-cancer therapy comprises administering to the subject a non-peptide molecule that forms a covalent bond with a target protein in a cancer cell of the subject, the method further comprising administering to the subject an effective amount of a binding partner that specifically binds to a peptide conjugate, wherein the peptide conjugate comprises a peptide derived from the target protein covalently bound to the non-peptide molecule, and wherein the binding partner mediates immune cell-mediated killing of the cancer cell or antibody-drug conjugate (ADC)-mediated killing, antibody-dependent cellular phagocytosis (ADCP) or complement-dependent cytotoxicity (CDC)-mediated killing of the cancer cell, or by using the binding partner as an immunotoxin or by using the binding partner as a radioconjugate in the subject. In some embodiments, the peptide conjugate is presented on the cancer cell of the subject in the context of an MHC molecule. In some embodiments, the binding partner specifically binds to a complex comprising the peptide conjugate and the MHC molecule. In some embodiments, the binding partner does not detectably bind to a complex of the peptide and the MHC molecule, wherein the peptide is not covalently bound to the non-peptide molecule. In some embodiments, the subject has previously received the non-peptide molecule prior to administration of the binding partner.

[0130] In another aspect, the present disclosure provides a kit comprising: (i) a non-peptide molecule, wherein the 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 the peptide conjugate comprises a peptide derived from the target protein covalently bound to the non-peptide molecule, and optionally (iii) instructions for use.

[0131] In another aspect, the present disclosure provides an isolated peptide conjugate comprising a peptide 7-30 amino acids in length comprising an amino acid sequence at least 80% identical to the amino acid sequence VVGACGVGK, wherein the peptide is conjugated to sotolacib or a derivative thereof.

[0132] In another aspect, the present disclosure provides an isolated peptide conjugate comprising a peptide having a length of 7-30 amino acids, said peptide comprising an amino acid sequence at least 80% identical to the amino acid sequence QLMPFGCLL, wherein said peptide is conjugated to osimertinib or a derivative thereof.

[0133] In another aspect, the present disclosure provides an isolated peptide conjugate comprising a peptide 7-30 amino acids in length comprising an amino acid sequence at least 80% identical to the amino acid sequence YMANGCLLNY, wherein the peptide is conjugated to ibrutinib or a derivative thereof.

[0134] In another aspect, the present disclosure provides an isolated molecular complex comprising a peptide conjugate disclosed herein and an MHC molecule, or a fragment or derivative thereof.

[0135] In another aspect, the present disclosure provides a host cell comprising the molecular complex disclosed herein.

[0136] In another aspect, the present disclosure provides a solid surface support comprising a molecular complex disclosed herein.

[0137] In another aspect, the present disclosure provides a kit comprising: (i) a peptide conjugate disclosed herein, a molecular complex disclosed herein, a cell disclosed herein, a solid surface support disclosed herein, or any combination thereof, and optionally (ii) instructions for use.

[0138] In another aspect, the present disclosure provides a kind of separated binding partner, and this binding partner specifically binds to the peptide conjugate disclosed herein or the molecular complex disclosed herein.In some embodiments, the binding partner is an antibody or its antigen-binding fragment.In some embodiments, the binding partner is a component of a chimeric antigen receptor (CAR).

[0139] In another aspect, the present disclosure provides a fusion protein comprising a binding partner disclosed herein, said fusion protein comprising (i) at least one cytokine, and / or (ii) an additional antibody or fragment thereof that is an immune checkpoint inhibitor. 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, and combinations thereof, and / or (ii) the additional antibody or fragment thereof that is an immune checkpoint inhibitor is an 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-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, or an agonist anti-CD28 antibody. In some embodiments, the binding partner comprises a component that binds to a T cell or natural killer (NK) cell protein, wherein the T cell or NK cell protein is selected from the group consisting of T cell receptor protein, CD4, CD8, CD28, CD16A, NKG2D, NKp30, NKp46, and combinations thereof.

[0140] 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, VH comprises a heavy chain complementary determining region 3 (CDR-H3), wherein the heavy chain complementary determining region 3 (CDR-H3) comprises an amino acid sequence of GX1WX2X3AMDY, wherein X1 is G, R, H, S or K, X2 is Y or I, and X3 is P or A. In some embodiments, VL comprises a light chain complementary determining region 3 (CDR-L3), wherein the light chain complementary determining region 3 CDR-L3 comprises an amino acid sequence of QQX1SYVX2X3X4IT, wherein X1 is I, A, P, V or S, X2 is K, R, A or H, X3 is K or R, and X4 is L, T, K, R, V, A or E.

[0141] 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 a fragment thereof; and wherein the polypeptide binds to an epitope of the MHC.

[0142] In another aspect, the 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 a fragment thereof; and wherein the antigen binding domain binds to the peptide conjugate / MHC complex with a dissociation constant (K D ) is at most about 50 nM.

[0143] In another aspect, the 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 a fragment thereof; and wherein the dissociation constant (K) of the antigen binding domain for binding to a free targeted covalent inhibitor is D ) is at least 200 nM.

[0144] 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 a fragment thereof; and wherein: (a) the polypeptide binds to the peptide conjugate / MHC complex at an angle of about 10° to 60° between the axis of the MHC and the axis of the polypeptide; (b) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is about 10° to 60°; (c) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is different from that when a TCR is bound to the peptide conjugate / MHC complex. (d) the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex when the polypeptide is bound to the peptide conjugate / MHC complex is about 10° to 60° or about 10° to 60° less than the angle between the axis of the TCR and the axis of the peptide conjugate / MHC complex when the TCR is bound to the peptide conjugate / MHC complex; (e) the polypeptide binds to the peptide conjugate / MHC complex at an angle that is different from the angle at which the T cell receptor binds to the peptide conjugate / MHC complex; and / or (f) the polypeptide binds to the peptide conjugate / MHC complex at an angle that is about 10° to 60° or about 10° to 60° less than the angle at which the T cell receptor binds to the peptide conjugate / MHC complex.

[0145] 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 a fragment thereof; and wherein the polypeptide contacts one or more residues of the α1 domain or region and one or more residues of the α2 domain or region of the heavy chain of the MHC of the peptide conjugate / MHC complex.

[0146] 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 a fragment thereof; and wherein the polypeptide binds to residues 62-66, 106-109 and / or 150-170 of MHC or one or more residues thereof.

[0147] In another aspect, the 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 a fragment thereof; and wherein the interfacial area of ​​the polypeptide with the peptide conjugate / MHC complex is at least about In some embodiments, the polypeptides disclosed herein comprise a heavy chain variable region (VH) and / or a light chain variable region (VL). In some embodiments, VH comprises a CDR-H3 comprising an amino acid sequence of GX1WX2X3AMDY, wherein X1 is G, R, H, S, or K, X2 is Y or I, and X3 is P or A. In some embodiments, VL comprises a CDR-L3 comprising an amino acid sequence of QQX1SYVX2X3X4IT, wherein X1 is I, A, P, V, or S, X2 is K, R, A, or H, X3 is K or R, and X4 is L, T, K, R, V, A, or E. In some embodiments, VH comprises: (i) a CDR-H1 comprising the amino acid sequence of DYSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) a CDR-H2 comprising the amino acid sequence of SISSSSGSTSYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and / or (iii) a CDR-H3 comprising the amino acid sequence of GX1WX2X3AMDY, wherein X1 is G, R, H, S, or K, X2 is Y or I, and X3 is P or A. In some embodiments, VL comprises: (i) a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) a CDR-L2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and / or (iii) a CDR-L3 comprising the amino acid sequence of QQX1SYVX2X3X4IT, wherein X1 is I, A, P, V, or S, X2 is K, R, A, or H, X3 is K or R, and X4 is L, T, K, R, V, A, or E.

[0148] In some embodiments, VH comprises a CDR-H1 having a sequence of DYSIH, a CDR-H2 having a sequence of SISSSSGSTSYADSVKG, and a CDR-H3 having a sequence of GGWIAAMDY. In some embodiments, VL comprises a CDR-L1 having a sequence of RASQSVSSAVA, a CDR-L2 having a sequence of SASSLYS, and a CDR-L3 having a sequence of QQASYVRKTIT. In some embodiments, VH comprises an amino acid sequence having at least about 90%, 95%, or 100% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYSIHWVRQAPGKGLEWVASISSSSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGWIAAMDYWGQGTLVTVSS. In some embodiments, VL comprises an amino acid sequence having at least about 90%, 95%, or 100% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0149] In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GSWIHAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SISSSWGVTSYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FHWYSIH. In some embodiments, the antigen binding domain further comprises a light chain complementary determining region 3 (VL), and wherein the VL comprises CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises: a CDR-H3 sequence of GSWIHAMDY, a CDR-H2 sequence of SISSSWGVTSYADSVKG, a CDR-H1 sequence of FHWYSIH, a CDR-L3 sequence of QQASYVRKTIT, a CDR-L2 sequence of SASSLYS, and a CDR-L1 sequence of RASQSVSSAVA. In some embodiments, the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFHWYSIHWVRQAPGKGLEWVASISSSWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIHAMDYWGQGTLVTVSS. In some embodiments, VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV. In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GHWIAAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SIASSSGSTGYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FSWYSIH.In some embodiments, the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises a light chain complementary determining region 3 (CDR-L3), and the light chain complementary determining region 3 (CDR-L3) comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises: the CDR-H3 sequence of GHWIAAMDY, the CDR-H2 sequence of SIASSSGSTGYADSVKG, the CDR-H1 sequence of FSWYSIH, the CDR-L3 sequence of QQASYVRKTIT, the CDR-L2 sequence of SASSLYS, and the CDR-L1 sequence of RASQSVSSAVA. In some embodiments, VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSWYSIHWVRQAPGKGLEWVASIASSSGSTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGHWIAAMDYWGQGTLVTVSS. In some embodiments, VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0150] In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GGVIHAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SILSRWGVTSYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FSPYSIH. In some embodiments, the antigen binding domain further comprises a light chain complementary determining region 3 (VL), and wherein the VL comprises CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises: a CDR-H3 sequence of GGVIHAMDY, a CDR-H2 sequence of SILSRWGVTSYADSVKG, a CDR-H1 sequence of FSPYSIH, a CDR-L3 sequence of QQASYVRKTIT, a CDR-L2 sequence of SASSLYS, and a CDR-L1 sequence of RASQSVSSAVA. In some embodiments, VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYSIHWVRQAPGKGLEWVASILSRWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGVIHAMDYWGQGTLVTVSS. In some embodiments, VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0151] In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GSWIAAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SISSWHGETGYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FSPYSIH. In some embodiments, the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises: a CDR-H3 sequence of GSWIAAMDY, a CDR-H2 sequence of SISSWHGETGYADSVKG, a CDR-H1 sequence of FSPYSIH, a CDR-L3 sequence of QQASYVRKTIT, a CDR-L2 sequence of SASSLYS, and a CDR-L1 sequence of RASQSVSSAVA. In some embodiments, the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYSIHWVRQAPGKGLEWVASISSWHGETGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS. In some embodiments, VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0152] In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GGWIAAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SISSLQGDTGYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FSWYSIH. In some embodiments, the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises a CDR-H3 sequence of GGWIAAMDY, a CDR-H2 sequence of SISSLQGDTGYADSVKG, a CDR-H1 sequence of FSWYSIH, a CDR-L3 sequence of QQASYVRKTIT, a CDR-L2 sequence of SASSLYS, and a CDR-L1 sequence of RASQSVSSAVA. In some embodiments, the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSWYSIHWVRQAPGKGLEWVASISSLQGDTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGWIAAMDYWGQGTLVTVSS. In some embodiments, VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0153] In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GSWIAAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SIASWYGDTGYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FHYYSIH. In some embodiments, the antigen-binding domain further comprises a light chain variable region (VL), and wherein the VL comprises CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises: a CDR-H3 sequence of GSWIAAMDY, a CDR-H2 sequence of SIASWYGDTGYADSVKG, a CDR-H1 sequence of FHYYSIH, a CDR-L3 sequence of QQASYVRKTIT, a CDR-L2 sequence of SASSLYS, and a CDR-L1 sequence of RASQSVSSAVA. In some embodiments, the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFHYYSIHWVRQAPGKGLEWVASIASWYGDTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS. In some embodiments, VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0154] In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GGRIEAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SISSWYGKTGYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FGYYSIH. In some embodiments, the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises a CDR-H3 sequence of GGRIEAMDY, a CDR-H2 sequence of SISSWYGKTGYADSVKG, a CDR-H1 sequence of FGYYSIH, a CDR-L3 sequence of QQASYVRKTIT, a CDR-L2 sequence of SASSLYS, and a CDR-L1 sequence of RASQSVSSAVA. In some embodiments, the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFGYYSIHWVRQAPGKGLEWVASISSWYGKTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGRIEAMDYWGQGTLVTVSS. In some embodiments, VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0155] In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GYWIEAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SIASSYGSTGYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FSKYSIH. In some embodiments, the antigen-binding domain further comprises a light chain variable region (VL), and wherein the VL comprises CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises a CDR-H3 sequence of GYWIEAMDY, a CDR-H2 sequence of SIASSYGSTGYADSVKG, a CDR-H1 sequence of FSKYSIH, a CDR-L3 sequence of QQASYVRKTIT, a CDR-L2 sequence of SASSLYS, and a CDR-L1 sequence of RASQSVSSAVA. In some embodiments, the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSKYSIHWVRQAPGKGLEWVASIAS SYGSTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYWIEAMDYWGQGTLVTVSS. In some embodiments, VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0156] In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GSWIAAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SIHSSIGTTGYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FGLYSIH. In some embodiments, the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises: a CDR-H3 sequence of GSWIAAMDY, a CDR-H2 sequence of SIHSSIGTTGYADSVKG, a CDR-H1 sequence of FGLYSIH, a CDR-L3 sequence of QQASYVRKTIT, a CDR-L2 sequence of SASSLYS, and a CDR-L1 sequence of RASQSVSSAVA. In some embodiments, VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFGLYSIHWVRQAPGKGLEWVASIHSSIGTTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS. In some embodiments, VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0157] In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GSVIHAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SILSWIGKTSYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FSPYSIH. In some embodiments, the antigen binding domain further comprises a light chain variable region (VL), and wherein VL comprises CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises: the CDR-H3 sequence of GSVIHAMDY, the CDR-H2 sequence of SILSWIGKTSYADSVKG, the CDR-H1 sequence of FSPYSIH, the CDR-L3 sequence of QQASYVRKTIT, the CDR-L2 sequence of SASSLYS, and the CDR-L1 sequence of RASQSVSSAVA. In some embodiments, the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYSIHWVRQAPGKGLEWVASILSWIGKTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSVIHAMDYWGQGTLVTVSS. In some embodiments, VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0158] In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GGWIAAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SIASRWGHTGYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FSPYHIH. In some embodiments, the antigen-binding domain further comprises a light chain variable region (VL), and wherein the VL comprises CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises: a CDR-H3 sequence of GGWIAAMDY, a CDR-H2 sequence of SIASRWGHTGYADSVKG, a CDR-H1 sequence of FSPYHIH, a CDR-L3 sequence of QQASYVRKTIT, a CDR-L2 sequence of SASSLYS, and a CDR-L1 sequence of RASQSVSSAVA. In some embodiments, VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYHIHWVRQAPGKGLEWVASIASRWGHTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGWIAAMDYWGQGTLVTVSS. In some embodiments, VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0159] In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GSWIAAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SIASLQGITGYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FHEYSIH. In some embodiments, the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises: a CDR-H3 sequence of GSWIAAMDY, a CDR-H2 sequence of SIASLQGITGYADSVKG, a CDR-H1 sequence of FHEYSIH, a CDR-L3 sequence of QQASYVRKTIT, a CDR-L2 sequence of SASSLYS, and a CDR-L1 sequence of RASQSVSSAVA. In some embodiments, VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFHEYSIHWVRQAPGKGLEWVASIASLQGITGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS. In some embodiments, VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0160] In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GGVIHAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SILSRWGVTSYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FSDYSIH. In some embodiments, the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises: a CDR-H3 sequence of GGVIHAMDY, a CDR-H2 sequence of SILSRWGVTSYADSVKG, a CDR-H1 sequence of FSDYSIH, a CDR-L3 sequence of QQASYVRKTIT, a CDR-L2 sequence of SASSLYS, and a CDR-L1 sequence of RASQSVSSAVA. In some embodiments, the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYSIHWVRQAPGKGLEWVASILSRWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGVIHAMDYWGQGTLVTVSS. In some embodiments, VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0161] In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GGVIHAMDY. In some embodiments, VH comprises CDR-H2, and the CDR-H2 comprises the amino acid sequence of SISSRWGVTSYADSVKG. In some embodiments, VH comprises CDR-H1, and the CDR-H1 comprises the amino acid sequence of FSDYSIH. In some embodiments, the antigen-binding domain further comprises a light chain variable region (VL), and wherein the VL comprises CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. In some embodiments, VL comprises CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS. In some embodiments, VL comprises CDR-L1, and the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. In some embodiments, the antigen binding domain comprises: a CDR-H3 sequence of GGVIHAMDY, a CDR-H2 sequence of SISSRWGVTSYADSVKG, a CDR-H1 sequence of FSDYSIH, a CDR-L3 sequence of QQASYVRKTIT, a CDR-L2 sequence of SASSLYS, and a CDR-L1 sequence of RASQSVSSAVA. In some embodiments, the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYSIHWVRQAPGKGLEWVASISSRWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGVIHAMDYWGQGTLVTVSS. In some embodiments, VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYSIHWVRQAPGKGLEWVASISSRWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGVIHAMDYWGQGTLVTVSS.

[0162] In some embodiments, the antibody or antigen-binding fragment specifically binds to the peptide conjugate / MHC complex, and wherein the antibody or 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 fragment thereof; and (b) an MHC. In some embodiments, the MHC is a human leukocyte antigen (HLA). In some embodiments, the HLA is an HLA-A*03:01, HLA-A*11:01, and / or HLA-A*02:01 molecule.

[0163] 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 (K D ) is 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 dissociation constant (K) of the antigen binding domain for binding to the peptide conjugate / MHC complex is D ) is from about 0.01 nM to about 20 nM. In some embodiments, the dissociation constant (K) for binding of the antigen binding domain to the free targeted covalent inhibitor is D ) is at least about 100 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 μM, at least about 10 μM, at least about 20 μM, at least about 30 μM, at least about 40 μM, at least about 50 μM, or at least 100 μM. In some embodiments, the dissociation constant (K) for binding of the antigen binding domain to a free targeted covalent inhibitor is D ) is the K of the antibody or antigen-binding fragment binding to the peptide conjugate / MHC complex D In some embodiments, the antigen binding domain binds undetectably to the free targeted covalent inhibitor. In some embodiments, the antigen binding domain binds undetectably to the free targeted covalent inhibitor with an IC of at least about 50 nM. 50 In some embodiments, the dissociation constant (K) of the antigen binding domain binding to the free peptide conjugate is D) is greater than about 100 nM, greater than about 200 nM, greater than about 300 nM, greater than about 400 nM, greater than about 500 nM, greater than 1 μM, greater than 10 μM, greater than 20 μM, greater than 30 μM, greater than 40 μM, greater than 50 μM, or greater than 100 μM. In some embodiments, the dissociation constant (K) of the antigen binding domain for binding to the free peptide conjugate is greater than about 100 nM, greater than about 200 nM, greater than about 300 nM, greater than about 400 nM, greater than about 500 nM, greater than 1 μM, greater than 10 μM, greater than 20 μM, greater than 30 μM, greater than 40 μM, greater than 50 μM, or greater than 100 μM. D ) is the K of the antibody or antigen-binding fragment and peptide conjugate / MHC complex D In some embodiments, the dissociation constant (K) of the antigen binding domain binding to the free peptide conjugate is at least about 10, 100, 10,000, 100,000 times greater than that of the antigen binding domain. D ) is the K of the antibody or antigen-binding fragment and peptide conjugate / MHC complex D In some embodiments, the dissociation constant (K) of the antigen binding domain binding to the free peptide conjugate is at least 2.5 times greater than that of the free peptide conjugate. D ) is greater than the K of the antibody or antigen-binding fragment and peptide conjugate / MHC complex D , wherein the MHC is HLA-A02:01. In some embodiments, the dissociation constant (K) of the antigen binding domain binding to the free peptide conjugate is D ) is the K of the antibody or antigen-binding fragment and peptide conjugate / MHC complex D In some embodiments, the antigen binding domain binds to the peptide conjugate / MHC complex with an affinity that is at least 100-fold, at least 200-fold, at least 300-fold, at least 400-fold, at least 500-fold, at least 600-fold, at least 700-fold, at least 800-fold, at least 900-fold, at least 1,000-fold, at least 2,500-fold, at least 5,000-fold, or at least 10,000-fold greater than the affinity of the antibody or antigen binding fragment for the free targeted covalent inhibitor or free peptide conjugate.

[0164] In some embodiments, the peptide conjugate consists of a free targeted covalent inhibitor and a KRAS G12C Peptide, KRAS G12D Peptide, KRAS G12R Peptide or KRAS G12SIn some embodiments, the free targeted covalent inhibitor is osimertinib, ibrutinib, neratinib, sotolacib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1, or BI 182391. In some embodiments, the peptide conjugate is formed by sotolacib and KRAS G12C In some embodiments, the peptide comprises or consists of the amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV.

[0165] In some embodiments, the antigen binding domain (i) is specific for a peptide conjugate / MHC complex comprising VVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*11:01 and / or a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*11:01, or (ii) is specific for a peptide conjugate / MHC complex comprising VVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:01 and / or a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:01. In some embodiments, the antigen binding domain is specific for a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:01 and / or a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*11:01. In some embodiments, the antigen binding domain is specific for a peptide conjugate / MHC complex comprising VVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*11:01, a peptide conjugate / MHC complex comprising VVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:01, a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:01, a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*11:01, and / or a peptide conjugate / MHC complex comprising KLVVVGACGV conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*02:01.

[0166] In some embodiments, the polypeptide binds to: (i) a peptide conjugate comprising a peptide consisting of the amino acid sequence VVGACGVGK / HLA-A*11:01 MHC complex and a peptide conjugate comprising a peptide consisting of the amino acid sequence VVVGACGVGK / HLA-A*11:01 MHC complex; (ii) a peptide conjugate comprising a peptide consisting of the amino acid sequence VVGACGVGK / HLA-A*03:01 MHC complex and a peptide conjugate comprising a peptide consisting of the amino acid sequence VVVGACGVGK / HLA-A*03:01 MHC complex; (iii) a peptide conjugate comprising a peptide consisting of the amino acid sequence VVGACGVGK / HLA-A*03:01 MHC complex (iv) a peptide conjugate / HLA-A*03:01MHC complex comprising a peptide consisting of the amino acid sequence VVGACGVGK and a peptide conjugate / HLA-A*11:01MHC complex comprising a peptide consisting of the amino acid sequence VVVGACGVGK and a peptide conjugate / HLA-A*11:01MHC complex comprising a peptide consisting of the amino acid sequence VVVGACGVGK; and / or (v) a peptide conjugate / HLA-A*02:01MHC complex comprising a peptide consisting of the amino acid sequence KLVVVGACGV.

[0167] In some embodiments, the polypeptide binds to: a peptide conjugate comprising a peptide consisting of the amino acid sequence VVGACGVGK / HLA-A*03:01 MHC complex, and a peptide conjugate comprising a peptide consisting of the amino acid sequence VVVGACGVGK / HLA-A*03:01 MHC complex, and a peptide conjugate comprising a peptide consisting of the amino acid sequence VVGACGVGK / HLA-A*11:01 MHC complex, and a peptide conjugate comprising a peptide consisting of the amino acid sequence VVVGACGVGK / HLA-A*11:01 MHC complex, and a peptide conjugate comprising a peptide consisting of the amino acid sequence KLVVVGACGV / HLA-A*02:01 MHC complex.

[0168] In some embodiments, the targeted covalent inhibitor has a chemical structure comprising C-R1, wherein C is a chemical moiety attached to R1 of any compound selected from the group consisting of

[0169]

[0170] and wherein R1 is selected from

[0171]

[0172] In some embodiments, the targeted covalent inhibitor forms a covalent bond with: (i) a cysteine ​​residue in a peptide comprising the amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV, (ii) an aspartic acid residue in a peptide comprising the amino acid sequence of VVVGADGVGK, VVGADGVGK, or KLVVVGADGV, (iii) a serine residue in a peptide comprising the amino acid sequence of VVVGASGVGK, VVGASGVGK, or KLVVVGASGV, or (iv) an arginine residue in a peptide comprising the amino acid sequence of VVVGARGVGK, VVGARGVGK, or KLVVVGARGV.

[0173] In some embodiments, the antigen binding domain of the polypeptide recognizes portion C of the targeted covalent inhibitor of the peptide conjugate / MHC complex.

[0174] In some embodiments, the antigen binding domain of the polypeptide recognizes the C portion of the targeted covalent inhibitor of the peptide conjugate / MHC complex, but does not recognize the R1 portion of the targeted covalent inhibitor of the peptide conjugate / MHC complex.

[0175] In some embodiments, the interfacial area of ​​the polypeptide and the peptide conjugate / MHC complex is at least about or In some embodiments, the interfacial area of ​​the polypeptide with the MHC of the peptide conjugate / MHC complex is greater than the interfacial area of ​​the polypeptide with the peptide or targeted covalent inhibitor.

[0176] In some embodiments, the polypeptide forms a binding pocket at the interface between the VH and VL domains to accommodate a targeted covalent inhibitor of the peptide conjugate / MHC complex.

[0177] In some embodiments, the polypeptide does not bind to the peptide conjugate / MHC complex via head-to-head coaxial interactions.

[0178] In some embodiments, the polypeptide binds to the peptide conjugate / MHC complex at an angle of about 10° to 60° between the axis of the MHC and the axis of the polypeptide. In some embodiments, when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is about 10° to 60°. In some embodiments, when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is different from the angle between the axis of the TCR and the axis of the peptide conjugate / MHC complex when the TCR is bound to the peptide conjugate / MHC complex. In some embodiments, when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is about 10° to 60° or approximately 10° to 60° less than the angle between the axis of the TCR and the axis of the peptide conjugate / MHC complex when the TCR is bound to the peptide conjugate / MHC complex. In some embodiments, the polypeptide binds to the peptide conjugate / MHC complex at an angle that is different from the angle at which a T cell receptor binds to the peptide conjugate / MHC complex. In some embodiments, the polypeptide binds to the peptide conjugate / MHC complex at an angle that is about 10° to 60° or approximately 10° to 60° less than the angle at which a T cell receptor binds to the peptide conjugate / MHC complex.

[0179] In some embodiments, the polypeptide binds to the peptide conjugate / MHC complex at an angle of about 40° between the axis of the MHC and the axis of the polypeptide. In some embodiments, when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is about 40°. In some embodiments, when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is about 40° or approximately 40° less than the angle between the axis of the TCR and the axis of the peptide conjugate / MHC complex when the TCR is bound to the peptide conjugate / MHC complex. In some embodiments, the polypeptide binds to the peptide conjugate / MHC complex at an angle of about 40° or approximately 40° less than the angle at which a T cell receptor binds to the peptide conjugate / MHC complex.

[0180] In some embodiments, the polypeptide contacts the α1 domain and α2 domain of the MHC heavy chain of the peptide conjugate / MHC complex.

[0181] In some embodiments, the polypeptide binds to an epitope of MHC, and wherein the epitope comprises one or more residues from a region comprising residues 62-66, 106-109, and / or 150-170 of MHC.

[0182] In some embodiments, the polypeptide binds to an epitope of MHC, and wherein the epitope comprises one or more residues from a region comprising residues 62-66, 106-109, and / or 150-170 of an HLA-A*03:01 molecule or an HLA-A*11:01 molecule.

[0183] In some embodiments, the polypeptide binds to an epitope of MHC, and wherein the epitope comprises 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 molecule.

[0184] In some embodiments, the VL domain of the antigen binding domain binds to an epitope of MHC, and wherein the epitope comprises 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 HLA-A*03:01.

[0185] In some embodiments, the polypeptide binds to an epitope of MHC, and wherein the epitope comprises one or more residues selected from the group consisting of residues 62, 65, 66, 150, 151, 154, 155, 157, and 158 of HLA-A*03:01.

[0186] In some embodiments, the VH domain of the antigen binding domain binds to an epitope of MHC, and wherein the epitope comprises one or more residues selected from the group consisting of residues 62, 65, 66, 150, 151, 154, 155, 157, and 158 of HLA-A*03:01.

[0187] In some embodiments, the polypeptide binds to an epitope of MHC, and wherein the epitope comprises 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 HLA-A*11:01.

[0188] In some embodiments, the VL domain of the antigen binding domain binds to an epitope of MHC, and wherein the epitope comprises 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 HLA-A*11:01.

[0189] In some embodiments, the polypeptide binds to an epitope of MHC, and wherein the epitope comprises one or more residues selected from the group consisting of residues 62, 65, 66, 151, 154, 155, and 158 of HLA-A*11:01.

[0190] In some embodiments, the VH domain of the antigen binding domain binds to an epitope of MHC, and wherein the epitope comprises one or more residues selected from the group consisting of residues 62, 65, 66, 151, 154, 155, and 158 of HLA-A*11:01. In some embodiments, the VH is connected to the VL by a linker. In some embodiments, the linker comprises (G4S) n or (S4G) n , wherein n is any integer between 1 and 10. In some embodiments, the linker comprises a glycine-serine-alanine linker G4SA3 or a glycine-serine linker (G4S)4.

[0191] In some embodiments, the polypeptide is a whole antibody, a bispecific antibody, a multispecific antibody, an antigen-binding (Fab) fragment, a Fab' fragment, a (Fab')2 fragment, a Fd, a Fv, a dAb, a single domain fragment or a single monomeric variable antibody domain, a dual affinity retargeting (DART) molecule, a diabody (Db), a single chain diabody (scDb), a single chain variable fragment (scFv), a bispecific T cell engaging antibody (BiTE), a bispecific killer cell engaging antibody (BiKE), a CrossMab, a camelid antibody, a trispecific binding partner, a chimeric antigen receptor (CAR), a monomer (also known as an Adnectin), a DARPin, an anticarrier protein, an affibody, or an antibody mimic protein. In some embodiments, the polypeptide is a bispecific antibody. In some embodiments, the bispecific antibody is a bispecific T cell engaging antibody (BiTE).

[0192] In some embodiments, the polypeptide further comprises a second antigen binding domain that binds to a T cell surface marker. In some embodiments, the T cell surface marker is CD3ε, CD3γ, CD3δ, CD3η, TCRɑ or TCRβ of TCR. In some embodiments, the antigen binding domain and the second antigen binding domain are connected by a linker. In some embodiments, the linker comprises (G4S) n or (S4G) n , where n is any integer between 1 and 10.

[0193] In some embodiments, the linker is configured such that: (a) the polypeptide binds to the peptide conjugate / MHC complex at an angle of about 10° to 60° between the axis of the MHC and the axis of the polypeptide; (b) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is about 10° to 60°; (c) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is different from the angle between the axis of the TCR and the axis of the peptide conjugate / MHC complex when the TCR is bound to the peptide conjugate / MHC complex; (d) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is different from the angle between the axis of the TCR and the axis of the peptide conjugate / MHC complex when the TCR is bound to the peptide conjugate / MHC complex; (e) the polypeptide binds to the peptide conjugate / MHC complex at an angle that is different from the angle at which the T cell receptor binds to the peptide conjugate / MHC complex; and / or (f) the polypeptide binds to the peptide conjugate / MHC complex at an angle that is less than about 10° to 60° or about 10° to 60° than the angle at which the T cell receptor binds to the peptide conjugate / MHC complex.

[0194] In some embodiments, polypeptide includes the first polypeptide chain comprising an antigen binding domain and the second polypeptide chain comprising a second antigen binding domain. In some embodiments, the first polypeptide chain or the second polypeptide chain is further fused with a cytokine or its fragment. In some embodiments, the cytokine includes IL-2, IL-7, IL-15, IL-12, IL-18 or IL-21, or interferon (IFN). In some embodiments, peptide conjugates / MHC complexes are presented on the cell surface. In some embodiments, cells express the peptide conjugates / MHC complexes of low copy number, and wherein the low copy number is each single cell 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.

[0195] In some embodiments, the peptide of the peptide conjugate / MHC complex is derived from an intracellular protein.

[0196] In some embodiments, the polypeptide is a binding partner disclosed herein.

[0197] In another aspect, the present disclosure provides a polypeptide comprising an antigen binding domain that specifically binds to a peptide conjugate / MHC complex, wherein the peptide conjugate / MHC complex comprises: (c) a peptide conjugate comprising a peptide covalently linked to a targeted covalent inhibitor or a fragment thereof; and (b) MHC.

[0198] In another aspect, the present disclosure provides a pharmaceutical composition comprising a polypeptide disclosed herein and a pharmaceutically acceptable carrier.

[0199] In another aspect, the present disclosure provides a method of treating cancer in a subject who has been treated with a free targeted covalent inhibitor, the method comprising administering to the subject a polypeptide disclosed herein or a pharmaceutical composition disclosed herein. In some embodiments, the subject is refractory to treatment with the free targeted covalent inhibitor.

[0200] In another aspect, the present disclosure provides a method of treating cancer in a subject, the method comprising administering to the subject a polypeptide disclosed herein or a pharmaceutical composition disclosed herein after or simultaneously with the administration of a small molecule drug.

[0201] In another aspect, the present disclosure provides a method of treating cancer in a subject, the method comprising administering to the subject a targeted covalent inhibitor, and administering to the subject a polypeptide disclosed herein or a pharmaceutical composition disclosed herein. In some embodiments, the polypeptide disclosed herein or the pharmaceutical composition disclosed herein is administered after or simultaneously with the administration of the targeted covalent inhibitor.

[0202] In another aspect, the present disclosure provides a computer-assisted method for identifying or designing potential polypeptides comprising an antigen-binding domain that 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 a fragment thereof, the method comprising: (a) providing Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and Figure 61 (i) coordinates of at least two atoms of the peptide conjugate / MHC complex, (b) providing a structure of a candidate polypeptide comprising an antigen binding domain for binding to the peptide conjugate / MHC complex, (c) fitting the structure of the candidate polypeptide to at least two atoms of the peptide conjugate / MHC complex, wherein the fitting comprises determining interactions between one or more atoms of the antigen binding domain of the candidate polypeptide and atoms of the peptide conjugate / MHC complex, and (d) selecting the candidate polypeptide if the candidate polypeptide is predicted to bind to the peptide conjugate / MHC complex.

[0203] In another aspect, the present disclosure provides a computer-assisted method for designing a potential polypeptide comprising an antigen-binding domain that 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 a fragment thereof, the method comprising: (a) providing a structure of a candidate polypeptide comprising an antigen-binding domain for binding to a peptide conjugate / MHC complex, (b) providing Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and Figure 61 The method further comprises: (a) selecting a candidate polypeptide comprising: (i) the coordinates of at least two atoms of the peptide conjugate / MHC complex and the coordinates of at least two atoms of the antigen-binding domain of the polypeptide bound to the peptide conjugate / MHC complex; (c) fitting the structure of the candidate polypeptide to the at least two atoms of the peptide conjugate / MHC complex and the at least two atoms of the antigen-binding domain of the polypeptide bound to the peptide conjugate / MHC complex, wherein the fitting comprises determining interactions between one or more atoms of the antigen-binding domain of the candidate polypeptide and atoms of the peptide conjugate / MHC complex, and (d) selecting the candidate polypeptide if the candidate polypeptide is predicted to bind to the peptide conjugate / MHC complex.

[0204] In another aspect, the present disclosure provides a computer-assisted method for designing potential polypeptides comprising an antigen-binding domain that 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 a fragment thereof, the method comprising: (a) providing a structure of a polypeptide comprising an antigen-binding domain that binds to the 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 the peptide conjugate / MHC complex. (d) selecting a polypeptide, wherein the antigen binding domain of the candidate polypeptide comprises one or more amino acid substitutions relative to a polypeptide comprising the antigen binding domain, wherein the one or more amino acid substitutions are residues in the antigen binding domain of the polypeptide comprising one or more atoms that interact with or modulate the interaction with 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 greater affinity than the affinity of the polypeptide comprising the antigen binding domain to the peptide conjugate / MHC complex.

[0205] In another aspect, the present disclosure provides a computer-assisted method for identifying or designing potential polypeptides comprising an antigen-binding domain that 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 a fragment thereof, the method comprising: using a computer system, for example, a programmed computer comprising a processor, a data storage system, an input device, and an output device, performing the following steps: (a) inputting a peptide comprising a peptide from or belonging to a target antigen; Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and 61 into a programmed computer, thereby generating a data set, (b) using the processor to compare the data set to a computer database of structures of polypeptides comprising an antigen binding domain stored in the computer data storage system, said polypeptides binding or putatively binding or expected to bind to a peptide conjugate / MHC complex, (c) using computer methods to select structures from the database that are likely to bind to certain peptide conjugate / MHC complexes, (d) using computer methods to construct models of the selected structures, and (e) outputting the selected structures to the output device; and optionally synthesizing one or more selected structures; and further optionally testing whether the synthesized selected structures bind to the peptide conjugate / MHC complex.

[0206] In another aspect, the present disclosure provides a computer-readable medium comprising: Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and Figure 61 Atomic coordinate data for the structure of any of the peptide conjugates, wherein the data define the three-dimensional structure of a polypeptide comprising an antigen binding domain bound to a peptide conjugate / MHC complex or provide the structure thereof, wherein the peptide conjugate of the peptide conjugate / MHC complex is a peptide covalently linked to a targeted covalent inhibitor or a fragment thereof or at least one subdomain thereof, or structure factor data for a polypeptide comprising an antigen binding domain bound to a peptide conjugate / MHC complex, wherein the structure factor data can be obtained from Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and Figure 61 The atomic coordinate data of any one of them is obtained.

[0207] In another aspect, the present disclosure provides a computer-readable medium comprising: Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and Figure 61Atomic coordinate data for the structure of any of the peptide conjugates / MHC complexes, said data defining the three-dimensional structure of or providing the structure of 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 a fragment thereof or at least one subdomain thereof, or structure factor data for the peptide conjugate / MHC complex, said structure factor data being available from Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and Figure 61 The atomic coordinate data of any one of the structures is obtained.

[0208] In another aspect, the present disclosure provides a method of identifying a T cell receptor (TCR) that recognizes a peptide conjugate / MHC complex disclosed herein, the method comprising: (a) contacting a plurality of candidate TCRs with the peptide conjugate / MHC complex, and (b) identifying at least one TCR that binds to the peptide conjugate / MHC complex.

[0209] In another aspect, the present disclosure provides a method for identifying a T cell receptor (TCR) that recognizes a peptide conjugate / MHC complex disclosed herein, the method comprising: (a) culturing T cells and antigen presenting cells (APCs) treated with a targeted covalent inhibitor to generate peptide conjugate / MHC complex-specific T cells having a T cell receptor (TCR) that binds to the peptide conjugate / MHC complex; and (b) identifying at least one TCR from the peptide conjugate / MHC complex-specific T cells that bind to the peptide conjugate / MHC complex. In some embodiments, the methods disclosed herein further comprise selecting or isolating at least one TCR.

[0210] In some embodiments, multiple candidate TCRs are multiple soluble TCRs or multiple TCRs expressed on the cell surface of multiple cells. In some embodiments, multiple candidate TCRs are multiple TCRs expressed on the cell surface of multiple cells, and the identification in (b) includes separating or selecting cells comprising at least one TCR based on cell activation markers. 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, CD137 or KLRG1.

[0211] In another aspect, the present disclosure provides a T cell receptor (TCR) comprising at least one TCR disclosed herein. In some embodiments, the TCR is a soluble TCR. In some embodiments, the TCR is a bispecific TCR.

[0212] In some aspects, disclosed herein is a method for treating cancer. In some embodiments, the method comprises prophylactically administering a peptide conjugate to a subject in need thereof before the subject develops cancer, before the subject is administered a drug, or both, wherein the peptide conjugate comprises a peptide covalently linked to a targeted covalent inhibitor or a fragment thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0213] Figure 1A-Figure 1C .Graph depicting phage enzyme-linked immunosorbent assay (ELISA) results of phage-displayed antibody clones. G12C -GDP and KRAS G12C -GDP-ARS-1620 conjugate binding. For each clone, the columns in the figure from left to right are buffer, G12C-GDP-ARS and G12C-GDP. The results of clones 1 to 32 are shown in Figure 1A As shown, the results of clone 33-clone 64 are as follows Figure 1B As shown, and the results of clone 65-clone 96 are as follows Figure 1C shown.

[0214] Figure 2 . Characterization of the binding of 12C-ARS Fab59 to the GTPγS- or GDP-bound nucleotide states of KRAS in the presence or absence of ARS-1620 using a bead binding assay G12C Combined diagram.

[0215] Figure 3 The figure shows that 12C-ARS-Fab59 specifically binds to KRAS conjugated with ARS-1620 G12C- GDP.

[0216] Figures 4A-4D . Figure 4A ARS-1620 / KRAS was measured using 12C-ARS-Fab59 in a pull-down assay using lysates prepared from cell lines. G12C Data obtained for the adduct. Figure 4B Shown are the results of experiments with the indicated KRAS mutants ( Figure 4A ) and KCP(Kras G12C ;Tp53 R172H ; Pdx-Cre) mouse pancreatic cancer cells ( Figure 4B ) Immunoblotting of whole-cell lysates and 12C-ARS Fab pull-downs (PD) of reconstituted RAS-null MEFs treated in the presence or absence of ARS-1620. Figure 4C Shown are whole cell lysates and 12C-ARS Fab pull-downs (PD) from H358 and MIAPaCa-2 cells, processed as indicated. Figure 4D The quantitative analysis by LC / MS-MS was shown. Figure 4C ARS adduct formation in samples. The concentration of ARS-1620 and SHP099 in all figures is 10 μM.

[0217] Figure 5A and Figure 5B The data show that 12C-ARS-Fab59 can be used to measure the engagement of ARS-1620 with mutant KRAS by a pull-down assay using lysates prepared from animal tissues. Figure 5A and Figure 5B showed that after oral gavage for 3 days of ARS-1620 (200 mg / kg / d) alone or together with the SHP2 inhibitor SHP099 (75 mg / kg / d), the LSL-KRAS G12C -Tp53 R270H ( Figure 5A ) and LSL-KRAS G12C ( Figure 5B ) Tumor lysates and 12C-ARS Fab pull-downs (PD) were immunoblotted with anti-pan-RAS and anti-ERK2 (loading control).

[0218] Figure 6 Binding of antibody clones to AMG-510 conjugated to KRAS (G12C) peptide and polyserine (irrelevant) peptide. For each antibody clone, the columns from left to right are no target, KRAS (G12C), KRAS (G12C)-AMG, and irrelevant peptide-AMG. The signals of the two negative controls (no target and KRAS (G12C) peptide without conjugated drug) were too low to be seen in the figure. The antibody clones were displayed on the surface of yeast cells, and flow cytometry was used to detect binding to targets conjugated to fluorescently labeled streptavidin.

[0219] Figure 7 Binding of the human IgG1 version of the P2AMR-1 clone to AMG-510 conjugated to the KRAS(G12C) peptide (circles) and the same peptide without drug conjugation (squares). The peptide was immobilized on streptavidin-coated beads, and the antibody bound to the beads was detected with a fluorescently labeled secondary antibody. The apparent K D Values. Data shown here are from three replicates. Error bars are within the symbol size.

[0220] Figure 8 Recognition of AMG-510 presented on MHC class I molecules. AMG510-RAS(G12C) conjugate (circles) and unbound peptide (squares) were loaded onto HLA-A*03:01 and immobilized on streptavidin-coated beads. Figure 2The same method was used to detect antibody binding. The apparent K D Values. Data shown here are from three replicates. Error bars are within the symbol size.

[0221] Figure 9 Called HapImmune TM A cartoon representation of the disclosed concept.

[0222] Figures 10A-10C Data representing the development of antibodies that bind to MHC / peptide-drug conjugate complexes. Figure 10A Shown is a multiplex bead binding assay (MBBA) of phage displaying different antibody clones. Figure 10B Shown are MBBA assays of phage displaying different antibody clones: HLA-A*01:01 complexed with BTK peptide conjugated to ibrutinib. Figure 10C Shown are MBBA assays of phage displaying different antibody clones: HLA-A*02:01 complexed with EGFR peptide conjugated to osimertinib.

[0223] Figures 11A-11C Figure 3. Binding titration of purified antibodies targeting the KRAS(G12C)-AMG510 conjugate using a multiplex bead binding assay (MBBA). The clone names are shown on each graph. Figure 11A Diagrams of clones AMRA3-7hIgG1, AMRA3-18hIgG1, and AMRA3-22hIgG1 are shown; Figure 11B Diagrams of clones AMRA11-2hIgG1 and AMRA11-15hIgG1 are shown; Figure 11C Maps of clones AMRA311-16hIgG1, AMRA311-17hIgG1, and AMRA311-18hIgG1 are shown. Antigen nomenclature is as shown in FIG10 .

[0224] Figure 12 The figure shows that the binding of the antibody to the AMG510-peptide conjugate complexed with HLA is not affected by the presence of free drug AMG-510.

[0225] Figure 13 Figure 10. Binding titration of purified antibodies targeting BTK-ibrutinib conjugate using a multiplex bead binding assay (MBBA). Clone names are shown on each graph. Antigen nomenclature is as in Figure 10.

[0226] Figure 14A and Figure 14BFigure 2. Binding titration of purified antibody to KRAS(G12C)-AMG510 conjugate presented by endogenous HLA molecules on the cell surface. Raji cells were first incubated with KRAS(G12C)-AMG510 conjugate or KRAS (wild-type) peptide, and then excess conjugate and peptide were washed away. Antibody levels detected using fluorescently labeled secondary antibodies are shown as a function of the IgG concentration used for staining. The apparent dissociation constant (K) was determined using a nonlinear least squares fit of a 1:1 binding function. D ) values. The data shown here are from three replicate measurements. Figure 14A Diagrams showing antibodies AMRA3-7 hIgG1, AMRA3-22 hIgG1, AMRA3-18 hIgG1, and AMRA11-2 hIgG1; Figure 14B Diagrams are shown for antibodies AMRA3-7hIgG1, AMRA3-22hIgG1, AMRA3-18hIgG1, and AMRA11-2hIgG1.

[0227] Figures 15A-15C The antibody binds to KRAS(G12C)-expressing cell lines pretreated with AMG-510. Figure 15A Flow cytometry histograms are shown. Figure 15B Shown are quantifications of median fluorescence intensity of H358 cells treated or not with AMG-510. Figure 15C Shown are quantifications of median fluorescence intensity of H358 cells and HEK293T cells treated or not with AMG-510 and stained with AMRA3-7 antibody (negative control).

[0228] Figure 16 Graph showing binding of P2AMR-1 IgG to cells pre-incubated with KRAS(G12C) peptide-AMG510 conjugate, KRAS (wild-type) peptide, or no peptide.

[0229] Figure 17A and Figure 17B Graph showing binding of purified antibodies in IgG format to the indicated drug-peptide / HLA complexes measured using a multiplex bead binding assay (MBBA). Figure 17A Shown are antibody clones identified with AMG-510 conjugated with KRAS(G12C) peptide complexed with HLA-A*03:01 as antigen. Figure 17B Shown are antibody clones identified with AMG-510 conjugated with KRAS(G12C) peptide complexed with HLA-*11:01 as antigen.

[0230] Figure 18A and Figure 18BThe figure shows the cytotoxic effect of single-chain diabody (scDb) on cells pulsed with exogenous peptide-drug conjugates. Figure 18A Shown are Raji cells that were first pulsed with AMG-510 conjugated to a peptide corresponding to a KRAS (G12C) fragment or a control peptide corresponding to KRAS (wild type). Pulsed cells were co-cultured with human T cells (effector: target = 3:1) in the presence of the indicated concentrations of single-chain diabodies (scDb). After incubation, dead cells were stained and detected by flow cytometry. The data shown here are from three replicates. Error bars represent standard deviations. When error bars are not visible, the error is less than the symbol. Figure 18B An equivalent experiment using T2 cells and osimertinib conjugated to an EGFR peptide is shown. As a negative control, a peptide conjugated to β-mercaptoethanol was used.

[0231] Figure 19A and Figure 19B The figure shows the specific cytotoxic effect of AMRA3-7_UCHT1 scDb on drug-treated lung cancer cell lines. Figure 19A Shown are lung cancer cell lines treated with 100 nM AMG-510 for 24 hours and then co-cultured with human T cells (E:T = 5:1) in the presence of AMRA3-7_UCHT1 scDb. After incubation, cell viability was measured. The scDb antibody exhibited a dose-dependent cytotoxic effect only against AMG510-treated cells harboring the cognate KRAS mutation (G12C) and HLA (HLA-A3). Figure 19B The cytotoxic effect of scDb at a concentration of 0.1 nM is shown. The data shown here are from four replicate measurements.

[0232] Figure 20 Figure 1 shows the binding titration curves of AMR-A3-7 and AMR-A3-7D displayed on the surface of yeast cells and HLA-A*03:01 presenting 9-mer or 10-mer RAS(G12C) peptides (VVGACGVGK and VVVGACGVGK, respectively) with AMG-510 conjugated to their Cys residues.

[0233] Figure 21A and Figure 21B . A graph showing the cell-killing effect of AMRA3-7D scDb. Figure 21A Shown is the dose-dependent cell killing effect of AMRA3-7D scDb tested using Raji cells, which were first pulsed with AMG-510 conjugated to a peptide corresponding to the KRAS(G12C) fragment (RAS-AMG510) or a control peptide corresponding to KRAS (wild-type, WTRAS). Figure 21BShown is the dose-dependent cell killing effect of AMRA3-7D scDb tested on the H2122 non-small cell lung cancer cell line treated with AMG-510 or DMSO alone (negative control).

[0234] Figures 22A-22C .Cell binding and killing effects of AMRA3-7D CrossMab. Figure 22A Binding of AMRA3-7D to CD3-expressing Jurkat cells and CD3-non-expressing Raji cells is shown. Figure 22B Shown is the dose-dependent cell killing effect of AMRA3-7D CrossMab tested using Raji cells, which were first pulsed with AMG-510 conjugated to a peptide corresponding to the KRAS(G12C) fragment (RAS-AMG510) or a control peptide corresponding to KRAS (wild-type, WTRAS). Figure 22C Shown are the cell killing effects of AMRA3-7DCrossMab tested on the H2122 non-small cell lung cancer cell line treated with AMG-510 or DMSO only (negative control).

[0235] Figure 23A and Figure 23B .Deep mutation scan of AMR-A3-7D. Figure 23A Shown are representative flow cytometry profiles of yeast cells displaying AMRA3-7D and its deep mutational scanning library derivatives. Figure 23B The incidence of mutations at each position in a sorted subset of the deep mutation scanning library is shown as a heat map.

[0236] Figure 24A and Figure 24B Deep mutational scanning of OEA2-5. Figure 24A Shown are representative flow cytometry profiles of yeast cells displaying OEA2-5 and its deep mutational scanning library derivatives as single-chain Fvs. Figure 24B The mutation incidence at each position in a sorted subset of the deep mutation scanning library is shown as a heat map.

[0237] Figure 25 Binding of single-chain Fv antibody clones to the AMG510-KRAS(G12C) peptide conjugate complexed with HLA-A*11 was measured in a yeast display format. Binding intensity is expressed as median fluorescence intensity (MEI) in arbitrary units.

[0238] Figures 26A-26F Selected clones tested in yeast display format with HLA-A*03 (top row; Figures 26A-26C ) and HLA-A*11 (bottom row; Figure 26D-26FBinding titration of the complexed AMG510-KRAS(G12C) peptide conjugate. Data for the 9-mer peptide VVGAC*GVGK (where C* represents the Cys residue conjugated to AMG-510) are shown as solid circles, and data for the 10-mer peptide VVVGAC*GVGK are shown as open circles. Data for the wild-type 9-mer VVGAGGVGK are shown as solid triangles. These curves are best fits to a 1:1 binding model.

[0239] Figures 27A-27D . Figures 27A-27C Shown are comparisons of the binding of antibody clones to the AMG510-KRAS(G12C) peptide (9-mer) presented on HLA-A*03 (filled circles) and to the isolated AMG-510-KRAS(G12C) peptide (i.e., in the absence of HLA-A*03) (open boxes). Figure 27D Binding of the antibodies to the AMG510-KRAS(G12C) peptide conjugate presented on HLA in the presence and absence of 10 μM free AMG-510 is shown, indicating that these antibodies are not inhibited by the presence of excess concentrations of free AMG-510.

[0240] Figure 28 .Cytotoxic effect of scDb antibodies on lung cancer cells treated with AMG510. Luciferase-expressing NCI-H2122 cells were incubated with 1 μM AMG-510 or DMSO (vehicle) and then co-cultured with human T cells (E:T = 10:1) in the presence of scDb. After incubation, target cell death was assessed by measuring luciferase activity released from dead cells into the culture medium. The affinity-matured antibodies showed potent cytotoxic effects in a manner that was dependent on AMG-510 treatment. The concentration of scDb was 10 nM, but 5 nM was used for anti-HLA-A3 scDb (positive control scDb targeting HLA-A*03). The data shown here are from four replicate measurements.

[0241] Figure 29 Cytotoxic effects of scDb antibodies on lung cancer cells treated with AMG510. Top, dose titration of three different scDb antibodies. Bottom, cytotoxic effects of 10 nM scDb compared to 5 nM control scDb. Figure 28 Proceed in the same way.

[0242] Figure 30 . Table E is shown, representing combinations of mutations that showed improved binding.

[0243] Figure 31 . Table F is shown, representing combinations of mutations that showed improved binding.

[0244] Figure 32 Binding titration of clones showing reduced affinity for the AMG510-KRAS(G12C) peptide conjugate complexed with HLA-A*03 relative to the affinity of the parental AMR3-7-D clone tested in a yeast display format. The experiment was performed in the same manner as Figure 26.

[0245] Figure 33 Shows the representative Figure 32 Table of mutation combinations showing reduced binding shown.

[0246] Figure 34 Shows the representative Figure 32 Table of mutation combinations showing reduced binding shown.

[0247] Figure 35 Biolayer interferometry (BLI) measurements of RA_D11 with AMG510-KRAS(G12C) peptide conjugates in complex with HLA-A*03 (left panel) and HLA-A*11 (right panel). Sensorgrams of HLA complexes with the indicated concentrations of the 9-mer peptide VVGAC*GVGK (where C* represents the Cys residue conjugated to AMG-510) are shown in black in the upper panel. Data for the 10-mer peptide VVVGAC*GVGK are shown in the lower panel. Data for HLA complexes with 64 nM of the wild-type 9-mer VVGAGGVGK and the 10-mer VVVGAGGVGK are shown in gray. RA_D11 in the form of a biotinylated Fab was immobilized on a BLI tip and reacted with HLA complexes in solution. D Values ​​were estimated by global curve fitting of a 1:1 binding model.

[0248] Figure 36 BLI sensorgrams showing binding of RA_D11 Fab to AMG510-KRAS(G12C) peptide conjugate in complex with HLA-A*02. Sensorgrams of HLA complexes with the indicated concentrations of the 10-mer peptide KLVVVGAC*GV (where C* represents the Cys residue conjugated to AMG-510). D Values ​​were estimated by global curve fitting of a 1:1 binding model.

[0249] Figure 37 Specific cytotoxicity of the RA_D11 scDb antibody against KRAS(G12C)-expressing H2122 (left panel) and SW1573 (right panel) lung cancer cell lines pretreated with 1 μM AMG-510. An anti-pan-HLA-A*03 scDb (A3-2) was used as a positive control. Note that it also binds to HLA-A02, although less tightly than HLA-A*03. Figure 28 Proceed in the same way.

[0250] Figure 38 As expected, the AMRA3-7 and RA_D11 scDb antibodies had no cytotoxic effect on wild-type KRAS-expressing Raji cells treated with AMG510. Raji cells were pretreated with 1 μM AMG-510 and stained with carboxyfluorescein succinimidyl ester (CSFE). Cells were then cocultured with human T cells (E:T = 5:1) in the presence of scDb for 18 hours, harvested, stained with the fixable viability dye eFluor 660, and analyzed by flow cytometry.

[0251] Figure 39 Binding titration of EO_Q16 and EO_Q17 to osimertinib-EGFR / HLA-A*02 (filled circles) and to EGFR / HLA-A*02 (no drug conjugation; open circles) showed that these antibodies have high affinity for their intended antigens but no affinity for the equivalent antibody without drug conjugation.

[0252] Figure 40 Binding of EO_Q16 and EO_Q17 to osimertinib-EGFR / HLA-A*02 (10 nM) in the absence and presence of free osimertinib (1 μM), indicating that antigen binding of these antibodies is not inhibited by the presence of free drug.

[0253] Figure 41 Cytotoxic effects of scDb antibodies on cells treated with drug-peptide conjugates. EO_Q16 and EO_Q17 scDb antibodies exhibited potent cytotoxic effects against cells treated with osimertinib-EGFRb peptide conjugates, but not against cells treated with the same peptide without drug conjugation. BB7.2 scDb, a pan-HLA-A2 clone, served as a positive control. Data shown are from triplicate measurements.

[0254] Figure 42 Cytotoxic effects of scDb antibodies on OCI-AML3 cells treated with osimertinib. EO_Q16 and EO_Q17 scDb antibodies showed no cytotoxic effects, as expected, as OCI-AML3 cells are not targeted by osimertinib. BB7.2 scDb, a pan-HLA-A2 clone, served as a positive control. Data shown are from triplicate measurements.

[0255] Figures 43A-43H .Cytotoxic effect of RA_D11 scDb on tumor cells treated with sotolacib. Figure 43A Shown are dose-response curves for the viability of H358 and H2122 cells after 72 hours of exposure to sotolacilib. Figure 43BWestern blot analysis of sotolacilib conjugation to KRAS(G12C) in H2122 cells is shown. H2122 cells were incubated with 100 nM sotolacilib for 24 hours. Arrows indicate KRAS(G12C) conjugated to sotolacilib. Note that the anti-pan RAS antibody detects KRAS, HRAS, and NRAS, so a complete shift of the original band is not expected. Figure 43C The cytotoxic effects of the indicated scDb on H2122-Nluc cells treated with 1 μM sotolacib are shown. The fold change in luminescent signal intensity is shown, normalized to the no scDb control (i.e., H2122 and T cells only). Except for A3-2 scDb (1 nM), the scDb concentration was 10 nM. DMSO is the solvent for sotolacib. Figure 43D Shown is the cell killing titration curve of RA_D11 scDb against H2122-Nluc cells treated with 1 μM sotolacib. Figure 43E The dependence of cell killing on sotolacib concentration at the indicated 1 nM scDb is shown. Figure 43F The HLA-dependence of cell killing by RA_D11scDb is shown. Normalized luminescence intensity is shown for cell lines treated with 0.3 μM sotolacib and co-cultured with T cells in the presence of 1 nM scDb and 0.3 μM sotolacib. The KRAS mutation status and HLA alleles of the cell lines are shown. Because each cell line expressed different Nluc levels, each assay was performed separately, and the relative luciferase level was determined for each cell line. Figure 43G and Figure 43H The cytotoxic effect of RA_D11 scDb (1 nM) on H2030-Nluc (G) and SW1573-Nluc (H) cells treated with sotolacib is shown. The data shown are from four replicate measurements.

[0256] Figure 44A and Figure 44B .HLA expression of cell lines. Figure 44A Analysis of HLA-A*03 expression on Raji cells is shown. Figure 44B Analysis of HLA-A*03 expression on H2122 cells and H2122(HLA-A*03KO) cells is shown. Note that the latter lacks HLA-A*03, indicating successful deletion.

[0257] Figures 45A-45D .The cytotoxic effect of RA_D11 on tumor cells treated with sotolacib. Figure 45AThe cytotoxic effects of the indicated scDb on H2122-Nluc cells treated with 1 mM sotolacib are shown. Except for the positive control A3-2 scDb (1 nM), the scDb concentration was 10 nM. Figure 45B Shown are the cytotoxic effects of the indicated scDb on various NLuc-expressing tumor cell lines treated with 0.3 μM sotolacib. The scDb concentration was 1 nM. Figure 45C and Figure 45D The results show the effect of RA_D11 scDb on H2030-Nluc cells treated with Sotolacib ( Figure 45C ) and SW1573-Nluc cells ( Figure 45D ) cytotoxicity. The scDb concentration was 1 nM. Anti-pan-HLA-A*03 / A*02 antibody (A3-2 scDb), anti-pan-HLA-A*02 antibody (BB7.2 scDb), and anti-pan-HLA-A*11 antibody (A11-1) served as positive controls. Data shown are from four replicate measurements.

[0258] Figure 46 Binding analysis of antibody phage clones to peptide / MHC complexes or sotolacib / peptide / MHC complexes by multiplex bead binding assay (MBBA). Phage MBBA signals demonstrated binding specificity for the antibody clones (A3-2 and A11-1) without being specific for the identity of the bound peptide.

[0259] Figures 47A-47C Structure of the .soto-pMHC-RA_D11 complex (side and top views). Figure 47A The side view structure (top panel) and top view structure (bottom panel) of soto-p8 / A03-RA_D11 are shown. Figure 47B The side view (top panel) and top view (bottom panel) structures of soto-p7 / A03-RA_D11 are shown. Figure 47C The side view (top figure) and top view (bottom figure) structures of soto-p7 / A11-RA_D11 are shown. Proteins and peptides are represented by tube diagrams, and soto-plasmid is represented by stick diagrams. 4701 represents the relative region of RA_D11 VH, 4702 represents the relative region of RA_D11 VL, 4703 represents the relative region of β2m, 4704 represents the relative region of HLA, and 4705 represents the relative position of the drug-peptide conjugate soto-p. The angle between the axis of FabRA_D11 and the axis of soto-pMHC is shown in the soto-p8 / A03 structure ( Figure 47A ).

[0260] Figure 48Figure 3. Surface area of ​​residues buried after interaction of Fab RA_D11 with soto-pMHC, calculated by PDBePISA. The gray shading represents the percentage of solvent-accessible residue surface buried after interaction of the indicated molecules, as indicated by the scale bar on the right (the darker the gray, the higher the percentage of buried surface area).

[0261] Figures 49A-49E Details of the interaction between Fab RA_D11 and SOTO-P8 / A03. Figure 49A Shown are soto-p8 (sticks; e.g., V8V9G 10 A 11 C 12 G 13 V 14 G 15 K 16 HLA-A*03:01 residues contacting Fab RA_D11 or sotolacib are represented as sticks. Figure 49B The contact areas between the HLA molecules and Fab RA_D11 are shown. The contact areas of Fab RA_D11 described on the surface of HLA-A*03:01 (4901 shows the approximate area of ​​contact with VL, 4902 shows the approximate area of ​​contact with VH), sotolacizumab (4903 shows the approximate area of ​​contact with sotolacizumab), or both (4904) are marked. Figure 49C A tubular representation of Fab RA_D11 in complex with soto-p8 / A03 (only soto-p8 / A03 is shown; sticks) is shown. Residues of Fab RA_D11 that contact HLA-A*03:01 or soto-p8 / A03 are shown as sticks. Figure 49D Shown are the contact areas of HLA-A*03:01 (4905), sotolacib (4906), or both (4907, area surrounding sotolacib) mapped on the surface of Fab RA_D11. Figure 49E HLA-A*03:01 residues (e.g. R 108 、D 161 、E 166 、W 167 、R 168 ) and Fab RA_D11 (eg Q 27 、S 29 、S 30 、S 31 、Y 93 ) Details of H-bond formation. Residues involved in H-bonding are shown as sticks, and H-bonds are shown as dashed lines.

[0262] Figures 50A-50E Interaction details of Fab RA_D11 binding to SOTO-p7 / A03. Figure 50A Shown are soto-p7 (sticks; e.g., V7V8V9G 10 A 11 C 12 G 13 V 14 G 15 K 16 HLA-A*03:01 residues contacting Fab RA_D11 or sotolacib are represented as sticks. Figure 50B Contact areas of Fab RA_D11 depicted on the surface of HLA-A*03:01 are shown (5001 shows the approximate area of ​​contact with VH; 5002: VL; 5003: VH and VL), sotolacib (5004), or both (5005). Figure 50C A tubular representation of Fab RA_D11 in complex with soto-p7 / A03 (only soto-p7 / A03 is shown; sticks) is shown. Residues of Fab RA_D11 that contact HLA-A*03:01 or soto-p7 / A03 are shown as sticks. Figure 50D Shown are the contact areas of HLA-A*03:01 (5006), sotolacib (5007), or both (5008) mapped on the surface of Fab RA_D11. Figure 50E The results show that the Fab RA_D11 VH (eg S33) and the Fab RA_D11 VL (eg Y 93 、S 92 、S 28 、S 30 )HLA-A*03:01 (e.g. E 166 、W 167 、R 108 ) residue details. Residues involved in hydrogen bonds are shown as sticks, and hydrogen bonds are shown as black dashed lines.

[0263] Figures 51A-51F Interaction details of Fab RA_D11 binding to SOTO-P7 / A11. Figure 51A Shown are soto-p7 (sticks; e.g., V8V9G 10 A 11 C 12 G 13 V 14 G 15 K 16HLA-A*11:01 residues contacting Fab RA_D11 or sotolacib are represented as sticks. Figure 51B Contact areas of Fab RA_D11 (5101 shows the approximate area of ​​contact with VH; 5102: VL; 5103: VH and VL), sotolacib (5104), or both (5105) depicted on the surface of HLA-A*11:01 are shown. Figure 51C A tubular representation of Fab RA_D11 in complex with soto-p7 / A11 is shown (soto-p7 / A11 only; sticks). Residues of Fab RA_D11 that contact HLA-A*11:01 or soto-p7 / A11 are shown as sticks. Figure 51D Shown are the contact areas of HLA-A*11:01 (5106), sotolacib (5107), or both (5108) mapped on the surface of Fab RA_D11. Figure 51E and Figure 51F Shown with Fab RA_D11 VH (eg G 100 、A 103 ) and RA_D11 VL (e.g. K 96 、S 92 、R 95 , Q 27 、S 28 , G 68 , R 66 ) forming H bonds between Sotolacizumab and HLA-A*11:01 (e.g. E 166 、R 169 、R 108 、D 106 ). Residues involved in H-bonding are shown as sticks, and hydrogen bonds are shown as black dashed lines.

[0264] Figures 52A-52F .Comparison of different Fab RA_D11-soto-pMHC structures. Figure 52A and Figure 52B A superposition of the SOTO-P8 / A03 and SOTO-P7 / A03 structures is shown. HLA-A*03:01 is represented by a tube. Side chains of HLA residues involved in the RA_D11 interaction are represented by sticks. Figure 52B Shown are comparative details between soto-p8 (dark) and soto-p7 (light) when loaded by HLA-A*03:01 (not shown) and bound to FabRA_D11 (not shown). Figure 52C-52FA superposition of the soto-p7 / A03 and soto-p7 / A11 structures is shown. Soto-p7 is represented by sticks (bright), while HLA-A*03:01 (bright) and HLA-A*11:01 (dark) are represented by tubes. The side chains of HLA residues that differ between HLA-A*03 and HLA-A*11 are represented by sticks. Figure 52D Shown are comparative details between soto-p7 peptide presented by HLA-A*03:01 (light bars, HLA not shown) or HLA-A*11:01 (dark bars, HLA not shown) and bound to Fab RA_D11 (not shown). Figure 52E Details of the differences in soto-p7 presentation caused by the presence of T163 / R163 in HLA-A*03 / A*11 are shown. Figure 52F Details are shown of the different conformations of Fab RA_D11 CDRH3 when bound to soto-p7 / A03 or soto-p7 / A11.

[0265] Figure 53 Flow cytometric profiles of RA_D11 (left column) bound to drug-peptide / MHC complexes and a sorted deep mutational scanning library. Condition 1: 10 nM soto-p7 / A03; Condition 2: 10 nM soto-p7 / A11; Condition 3: 10 nM soto-p5 / A02 tetramer (on streptavidin-DyLight650).

[0266] Figure 54 Heatmap representation of single-point mutants in the VL of RA_D11 that retained antigen binding (left column) or lost binding (right column). Numbers represent the number of reads for the mutation of interest divided by the total number of reads at that position multiplied by 100. Condition 1: 10 nM soto-p7 / A03; Condition 2: 10 nM soto-p7 / A11; Condition 3: 10 nM soto-p5 / A02 tetramer (on Streptavidin-DyLight650).

[0267] Figure 55 Heatmap representation of single-point mutants in RA_D11 VH that retained antigen binding (left column) or lost binding (right column). Numbers represent the number of reads for the mutation of interest divided by the total number of reads at that position multiplied by 100. Condition 1: 10 nM soto-p7 / A03; Condition 2: 10 nM soto-p7 / A11; Condition 3: 10 nM soto-p5 / A02 tetramer (on Streptavidin-DyLight650).

[0268] Figure 56The Fab RA_D11 binds specifically and with high affinity to all four soto-pMHC targets, and this binding is minimally inhibited by the free inhibitor. Minimal inhibition of RA_D11 binding by free soto-pMHC may be an essential prerequisite for co-administration with an inhibitor.

[0269] Figures 57A-57C CryoEM structures of RA_D11 bound to multiple drug-peptide / MHC complexes elucidated the molecular basis of target recognition. RA_D11 recognized the soto-p / A03-A11 target in a similar manner, primarily interacting with the HLA surface and soto-peptide (with a pocket formed by the interface of the Fab variable chain), while making minimal contacts with the rest of the peptide (p7 or p8).

[0270] Figure 58A and Figure 58B RA_D11 binds to soto-pMHC in a non-canonical manner, distinct from canonical TCR-pMHC engagement.

[0271] Figures 59A-59C Due to the different peptide adaptations of SOTO-P8 (stretched) and SOTO-P7 (bent) into the HLA-A*03 pocket, RA_D11 can recognize SOTO-P7 / A03 and SOTO-P8 / A03 in an almost identical manner. When the SOTO-P7 peptide is loaded by HLA-A*03 or HLA-A*11, the different residues between the two HLAs lead to slightly different conformations of the Fab CDRs (especially CDR-H3) upon target binding.

[0272] Figure 60A and Figure 60B RA_D11 can also bind to soto-p5 / A02, so specific T cell-engaging antibodies based on RA_D11 can also selectively kill KRAS with the most diffuse HLA-A*02. (G12C) RA_D11 binds soto-p5 / A02 in a non-uniform manner, displaying multiple possible orientations, and has reduced surface binding to HLA relative to soto-p / A03-A11 target binding.

[0273] Figure 61 .Soto-p5 is the same as soto-p 7-8 Very different peptides, and must be presented differently.

[0274] Figure 62Create a grid on the MHC backbone. Set the grid angles using the following α-carbons from the amino acids in the MHC complex: the origin is set to Met98; the X axis is set using Val28; the Y axis is set using Glu128; and the Y axis is orthogonal. The incident angle is measured along the XY axes, and the back angle is measured along the YZ axes.

[0275] Figure 63 Binding angles of soto-p7 / A03 and RA_D11 antibody. These angles are measured from the axis to the center of the RA_D11 antibody (Met105 sulfur). The back angle measured from the YZ axis was 64.7°. The incident angle measured from the XY axis was 77.3°.

[0276] Figure 64 Binding angles of soto-p7 / All and RA_D11 antibody. These angles are measured from the axis to the center of the RA_D11 antibody (Met105 sulfur). The back angle measured from the YZ axis was 67.2°. The incident angle measured from the XY axis was 78.4°.

[0277] Figure 65 Binding angles of soto-p8 / A03 and RA_D11 antibody. These angles are measured from the axis to the center of the RA_D11 antibody (Met105 sulfur). The back angle measured from the YZ axis was 65.1°. The incident angle measured from the XY axis was 77.2°.

[0278] Figure 66 Binding angles of the TCR-like antibody from PDB 3CHV. These angles are measured from the axis to the center of the TCR-like antibody (Phe107 C4). The back angle measured from the YZ axis is 92.5°. The incident angle measured from the XY axis is 111.1°. DETAILED DESCRIPTION

[0279] 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.

[0280] Every numerical range given throughout this specification will include its upper and lower numerical limits, and every narrower numerical range therein, as if such narrower numerical ranges were all expressly written herein.

[0281] As used in the specification and the appended claims, the singular forms "a," "an," 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 from particular value and / or to the other particular value. Similarly, when a value is expressed as an approximation by use of the preposition "about," it will be understood that the particular value forms another embodiment. The term "about" in relation to a numerical value includes variations of + / - 10%, + / - 5%, or + / - 1%.

[0282] The present disclosure includes each amino acid sequence described herein and all nucleotide sequences encoding amino acid sequences. Includes each antibody sequence and its antigen-binding fragment. The present invention includes polynucleotides and amino acid sequences with 80-99% similarity (including the end values ​​and including all values ​​and ranges therebetween) to the sequences provided herein. All amino acid sequences described herein may include amino acid substitutions, such as conservative substitutions, which do not adversely affect the function of the protein comprising the amino acid sequence. In this regard, the present disclosure provides alternative residues at certain positions in the binding partners, as described below. In certain instances, alternative residues are identified by deep mutation scanning, which demonstrates the binding function of each binding partner comprising the amino acid changes. The present disclosure includes each binding partner, wherein each alternative residue replaces the original residue alone and in any combination with the alternative residues. Therefore, any binding partner described herein may have any single residue change or combination of the changes. Representative changes for specific antibodies are described in the table. Changes may occur in CDR1, CDR2, CDR3 and combinations thereof. Changes may also include amino acid insertions. The present disclosure includes each amino acid sequence described by reference to the specific sequence identifier and the alternative amino acid described in the above table.

[0283] As described above, the present disclosure provides antibodies and antigen-binding fragments thereof (collectively referred to as "binding partners", and each individually referred to as a "binding partner"). The term "antibody" includes each binding partner form herein. The binding partner specifically binds to a protein or fragment thereof comprising a covalently linked molecule or a peptide provided in peptide form. The covalently linked molecules form a peptide conjugate. As used herein, "peptide conjugate" refers to any protein or peptide that has been modified so that it is covalently conjugated to another molecule. Peptide conjugates are considered to be a new type of antigen, i.e., a neoantigen. There are no particular restrictions on other molecules that are covalently conjugated to proteins or peptides to form peptide conjugates, provided that the other molecules are not additional amino acids added to the peptide conjugate. In some embodiments, the molecule covalently conjugated to a protein or peptide has or has had biological activity before conjugation, or may be biologically inert before conjugation. In some embodiments, the molecule is a drug, including but not necessarily limited to a small molecule drug. As used herein, molecules that are covalently linked to peptides to form peptide conjugates are referred to as "targeted covalent inhibitors (TCIs)" or "covalent drugs". In some cases, the targeted covalent inhibitor can be a molecule that inhibits the activity of the target protein, and can be covalently linked to a peptide from the target protein. In some cases, the targeted covalent inhibitor may not inhibit the activity of the target protein, but can be covalently linked to a peptide from the target protein. The representative and non-limiting examples of the medicine covalently linked to form a peptide conjugate with a peptide or protein 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, which include covalent modifications and, for example, produced by intracellular processing. In certain embodiments, the full-length protein can be covalently modified and subsequently processed in the cell, thereby producing a peptide conjugate as a fragment of the full-length protein. In one embodiment, the peptide conjugates include fragments of the full-length protein. As further described below, the peptide conjugates produced can be displayed on the cell surface. The cell surface display of the peptide conjugates can be any form of cell surface display, including but not limited to any receptor with an extracellular fragment, or can be displayed by any type of major histocompatibility complex (MHC) or human leukocyte antigen (HLA). Non-limiting examples of HLA types to which the peptide conjugates are displayed and to which the binding partner specifically binds are described further below.

[0284] 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 that is presented by the major histocompatibility complex (MHC). For example, the peptide conjugate can be formed by covalent reaction of the targeted covalent inhibitor with a residue (e.g., a cysteine ​​residue) in the peptide. In some embodiments, the peptide conjugate is composed of AMG-510 and KRAS G12CIn some embodiments, the peptide is introduced from the outside as a vaccine. In some embodiments, the peptide comprises a nucleophilic or electrophilic residue. In some embodiments, the residue comprises cysteine, lysine, tyrosine, histidine, serine, arginine or threonine. In one embodiment, the MHC is a human leukocyte antigen (HLA). In one embodiment, HLA is HLA-A*01:01, HLA-A*02:01, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*26:01, HLA-B*07:02, HLA-B*08:01, HLA-B*27:05, HLA-B*39:01, HLA-B*40:01, HLA-B*58:01 or HLA-B*15:01.

[0285] As used herein, the term "CDR" or "complementarity determining region" refers to the non-contiguous antigen binding sites found within the variable regions of heavy and light chain polypeptides. These specific regions have been described, for example, by 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 incorporated herein by reference in their entirety, wherein the definitions include overlapping or subsets of amino acid residues when compared to each other. In certain embodiments, the term "CDR" is a CDR as defined in MacCallum et al., J. Mol. Biol. 262: 732-745 (1996) and Martin A. "Protein Sequence and Structure Analysis of Antibody Variable Domains," Antibody Engineering, Kontermann and Dübel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001). In certain embodiments, the term "CDR" is a CDR as defined in Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest (1991). In certain embodiments, the heavy chain CDR and light chain CDR of an antibody are defined using different conventions. In certain embodiments, the heavy chain CDRs and / or light chain CDRs are defined by structural analysis of the antibody and identification of residues in the variable region predicted to contact an epitope region of a target molecule (e.g., a peptide conjugate). CDR-H1, CDR-H2, and CDR-H3 represent heavy chain CDRs, while CDR-L1, CDR-L2, and CDR-L3 represent light chain CDRs.

[0286] 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 for comparing two sequences is the algorithm of Karlin S & Altschul SF (1990) PNAS 87:2264-2268, as modified in Karlin S & Altschul SF (1993) PNAS 90:5873-5877, each of which is incorporated herein by reference in its entirety. This algorithm is incorporated into the NBLAST and XBLAST programs of Altschul SF et al., (1990) J Mol Biol 215:403, which is incorporated herein by reference in its entirety. BLAST nucleotide searches can be performed using the NBLAST nucleotide program parameter set, e.g., score = 100, wordlength = 12, to obtain nucleotide sequences homologous to the nucleic acid molecules described herein. BLAST protein searches can be performed using the XBLAST program parameter set, e.g., score 50, word length = 3, to obtain amino acid sequences homologous to the protein molecules described herein. To obtain gapped alignments for comparison purposes, Gapped BLAST can be used as described in Altschul SF et al., (1997) Nuc Acids Res 25: 3389-3402, which is incorporated herein by reference in its entirety. Alternatively, PSIBLAST can be used to perform an iterated search to detect distant relationships between molecules (Id.). When using BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used (see, e.g., the National Center for Biotechnology Information (NCBI) on the World Wide Web, ncbi.nlm.nih.gov). Another specific, non-limiting example of a mathematical algorithm used for sequence comparison is the algorithm of Myers and Miller, 1988, CABIOS 4: 11-17, which is incorporated herein by reference in its entirety. This algorithm is incorporated into the ALIGN program (version 2.0), which is a part of the GCG sequence alignment software package. When using the ALIGN program to compare 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 the two sequences can be determined using techniques similar to those described above, with or without gaps. When calculating percent identity, only exact matches are typically calculated.

[0287] As used herein, the term "free targeted covalent inhibitor" or "free drug" refers to a targeted covalent inhibitor that is not covalently attached to a protein or peptide. Once the targeted covalent inhibitor is covalently attached to a protein or peptide, the targeted covalent inhibitor may be referred to as a portion or fragment of the free targeted covalent inhibitor or drug. For example, a protein or fragment thereof may be a chemical fragment that binds to a cysteine ​​residue of a peptide after a covalent reaction between the free drug and the cysteine ​​residue of the peptide. As used herein, the term "KRAS G12C " refers to a KRAS protein with a G12C mutation (UniProt accession number P01116), i.e., a cysteine ​​residue at amino acid position 12. As used herein, the term "KRAS G12D " refers to a KRAS protein with a G12D mutation (UniProt accession number P01116), i.e., aspartic acid at amino acid position 12. As used herein, the term "KRAS G12R " refers to a KRAS protein with a G12R mutation (UniProt accession number P01116), i.e., an arginine at amino acid position 12. As used herein, the term "KRAS G12S ” refers to the KRAS protein with the G12S mutation (UniProt accession number P01116), i.e., serine at amino acid position 12.

[0288] In some embodiments, when covalently bound to a peptide conjugate, the binding partner preferentially binds to a protein or peptide or a complex comprising a protein or peptide relative to the same protein or peptide not bound to the drug. Thus, in the absence of a covalently linked molecule, the binding partner described herein does not detectably bind or binds to the same protein or fragment thereof with lower affinity. In some embodiments, the affinity of the binding partner to a protein or peptide comprising a covalently linked drug is 10-10,000 times, including all numbers and ranges between 10-10,000, of the affinity for a protein or peptide that does not comprise a covalently linked molecule. In this regard, and without intending to be bound by any particular theory, it is believed that the presence of a covalently linked molecule contributes to the epitope to which the binding partner specifically binds. Similarly, relative to binding to free drug, the binding partner of the present disclosure preferentially binds to the peptide conjugate. In some embodiments, the binding partner binds to a peptide comprising a covalently linked drug (e.g., a peptide conjugate / MHC complex) with an affinity that is 10-10,000 times greater than the affinity for the free drug, including all numbers and ranges therebetween. 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 100x, 1,000x, 10,000x, 100,000x, or more excesses of the free drug.

[0289] In some embodiments, the molecule covalently bound to form a peptide conjugate is a drug and can be any targeted covalent inhibitor (TCI), but a covalent drug does not necessarily inhibit the target peptide. In some embodiments, the molecule reacts with a specific residue within the target protein. In some embodiments, the molecule reacts at least partially with a protein or peptide fragment comprising a nucleophilic residue or an electrophilic residue. In some embodiments, the fragment of the protein or peptide to which the molecule reacts comprises any one 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, the disclosure of which is incorporated herein by reference. In some embodiments, the protein or peptide comprises selenocysteine. In some embodiments, the targeted covalent inhibitor reacts with selenocysteine. In some embodiments, the molecule reacts at least partially with a fragment of a protein or peptide comprising a wild-type Cys or a residue mutated to Cys and can thus be covalently linked via a so-called sulfur tether. In embodiments, the drug is any of the drugs 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 DeCesco, et al., European Journal of Medicinal Chemistry 138(2017)96e114, or Bauer, RA, Drug Discovery Today, Volume 20, Number 9, September 2015, wherein the disclosure of compounds that covalently modify protein targets is incorporated herein by reference.

[0290] In non-limiting embodiments, any of the Cys, Lys, Tyr, Ser, Thr, Arg, and His amino acids are present in the protein or peptide to which the molecule binds because the gene encoding the wild-type protein has mutated to encode a protein comprising one or a combination of said residues. In non-limiting embodiments, the molecule binds to a protein or peptide associated with a disease or condition (e.g., cancer, autoimmune disease, or other disease or disorder treated with a targeted covalent inhibitor). In some 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 fragment. In some embodiments, the drug binds to an enzyme that is not necessarily a receptor, including but not limited to any kinase. In some embodiments, the protein target includes a receptor with one or more activating mutations that promotes ligand-independent enzymatic activity.

[0291] In some embodiments, the molecule targets and thus covalently binds to an amino acid sequence present in any of the following proteins and / or variants thereof, which may or may not contain a mutation, such as a mutation associated with a particular disorder (including but not limited to any type of cancer). In some 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, the description of which is incorporated herein by reference. Visscher et al. also teach methods for identifying disease-associated mutant genes that introduce Cys residues suitable for covalent modification. In some embodiments, the protein is KRAS, Bruton's tyrosine kinase (BTK), any member of the epidermal growth factor receptor (EGFR) family (also known as the ERBB family), including but not limited to EGFR (ERBB1), HER2 / NEU (ERBB2), HER3 (ERBB3), and HER4 (ERBB4); fibroblast growth factor receptor (FGFR); receptor kinase known in the art as MET, BRAF, cyclin-dependent kinase (CDK); acetylcholinesterase (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 associated with obesity, such as pancreatic lipase and METAP2, or any cancer testis antigen. In some embodiments, the drug targets and thus covalently binds to any viral protein, including but not limited to polymerases, including any viral DNA polymerase, RNA polymerase, reverse transcriptase or RNA-dependent RNA polymerase, or a desired viral protein, such as a viral cell entry, or a protein encoded by any transposable element. In some embodiments, the drug targets EGFR and can be selected from PD168393, PF00299804 (dacomitinib), EKB569 (pelitinib), afatinib, WZ4002, osimertinib (formerly known as AZD9291), PF-06459988, nazartinib, naquintinib, omotinib, avitinib and rociterinib, neratinib, pyrotinib, poziotinib and derivatives thereof. In some embodiments, the drug targets Bruton's tyrosine kinase (BTK) and can be selected from ibrutinib, acalabrutinib, zanubrutinib, CHMFL-BTK-11, ONO / GS-405, PRN1008, and CC-292. In some embodiments, the drug targets any p90 ribosomal S6 kinase (RSK) and can be selected from fluoromethyl ketone (FMK) and dimethyl fumarate.In some embodiments, the drug targets any FGFR and can be selected from FIIN-1, FIIN-2, FIIN-3, BGJ398, AZD4547, PRN1371, FGF401. In some embodiments, the targeted covalent inhibitor targets E3 ligases such as RNF4, HOIP, RSP5, SMURF1, E6AP, HUWE1, and NEDD4-1. In one embodiment, the targeted covalent inhibitor targets DDB1 and CUL4-associated factors (DCAFs), such as DCAF1 or DCAF15. In one embodiment, the targeted covalent inhibitor targets any cancer testis antigen, any endogenous retroviral protein, long interspersed element-1 (LINE-1), or short interspersed element (SINE). In one embodiment, the targeted covalent inhibitor targets a short interspersed element that is optionally Alu. In one embodiment, the targeted covalent inhibitor is iniparib, abiraterone, carfilzomib, afatinib, or neratinib.

[0292] In some embodiments, the molecule 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 the residue numbering is the same for the relevant portions of all RAS isoforms discussed in this disclosure, and their amino acid sequences can be obtained, for example, from 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 G12 is the twelfth amino acid in the expressed RAS peptide sequence disclosed herein.

[0293] In one embodiment, the molecule covalently binds to a KRAS protein or peptide comprising a mutation. In some embodiments, the mutation is at least one of KRAS residues 12, 13, or 61. Reference herein to any drug includes both capitalized and non-capitalized forms of the drug.

[0294] In some embodiments, the drug targets a gene comprising KRAS G12C In some embodiments, the drug targets a KRAS protein. G12D In some embodiments, the drug targets a KRAS protein. G12R In some embodiments, the drug targets a KRAS protein. G12SMutated KRAS protein. In non-limiting embodiments, the drug targeting KRAS protein is selected from AMG-510 (sotolacib), ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1, or BI 182391, or derivatives thereof. In other embodiments, the drug targeting KRAS protein is selected from β-lactone G12Si-1, G12Si-2, G12Si-3, G12Si-4, and G12Si-5. In one embodiment, the drug comprises a proteolysis targeting chimera (PROTAC) derivative of a covalent drug, a non-limiting description of which can be found in doi: 10.1021 / acscentsci.0c00411, wherein the description of PROTAC is incorporated herein by reference. In some embodiments, PROTAC is LC-1 or LC-2. In some embodiments, the disclosure relates to autophagy-mediated degraders, referred to as AUTACs, as described in doi.org / 10.1080 / 15548627.2020.1718362, wherein the description of AUTACs is incorporated herein by reference.

[0295] The peptide conjugates described herein can be formed by covalent reaction of a free targeted covalent inhibitor with a KRAS peptide. G12C Peptide, KRAS G12D Peptide, KRAS G12R Peptide or KRAS G12S The free targeted covalent inhibitor can be any free targeted covalent inhibitor described herein. For example, the free targeted covalent inhibitor can be osimertinib, ibrutinib, neratinib, AMG-510, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1, BI 182391, or a derivative thereof. In other embodiments, the free targeted covalent inhibitor can be the β-lactone G12Si-1, G12Si-2, G12Si-3, G12Si-4, or G12Si-5. (See Table R). In some cases, the peptide conjugate can be conjugated to KRAS via AMG-510.G12C The peptide may comprise or consist of the amino acid sequence of VVVGACGVGK, VVGACGVGK or KLVVVGACGV.

[0296] The antigen binding domain of a polypeptide described herein or a 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 be specific for: (i) a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or fragment thereof presented by HLA-A*11:01, (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 be specific for: (i) a peptide conjugate / MHC complex comprising VVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:01, (ii) a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:01, or (iii) both. In other embodiments, the antigen binding domain can be specific for: (i) a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*11:01, (ii) a peptide conjugate / MHC complex comprising VVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*11:01, (iii) a peptide conjugate / MHC complex comprising VVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:01, Peptide conjugate / MHC complex, (iv) a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:01, (v) a peptide conjugate / MHC complex comprising KLVVVGACGV conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*02:01, or any combination of (i)-(v) (e.g., (i) and (ii); or (i), (ii) and (iv)), or all of (vii)-(i)-(v).In certain embodiments, the polypeptide binds to: (i) a peptide conjugate comprising a peptide consisting of the amino acid sequence VVGACGVGK / HLA-A*11:01 MHC complex, (ii) a peptide conjugate comprising a peptide consisting of the amino acid sequence VVVGACGVGK / HLA-A*11:01 MHC complex, (iii) a peptide conjugate comprising a peptide consisting of the amino acid sequence VVGACGVGK / HLA-A*03:01 MHC complex, (iv) a peptide conjugate comprising a peptide consisting of the amino acid sequence VVVGACGVGK / HLA-A*03:01 MHC complex, and / or (v) a peptide conjugate comprising a peptide consisting of the amino acid sequence KLVVVGACGV / HLA-A*02:01 MHC complex. In other embodiments, the polypeptide binds to: (i) a peptide conjugate comprising a peptide consisting of the amino acid sequence VVVGACGVGK / HLA-A*03:01 MHC complex and a peptide conjugate comprising a peptide consisting of the amino acid sequence VVVGACGVGK / HLA-A*11:01 MHC complex; (ii) a peptide conjugate comprising a peptide consisting of the amino acid sequence VVGACGVGK / HLA-A*03:01 MHC complex and a peptide conjugate comprising a peptide consisting of the amino acid sequence VVVGACGVGK / HLA-A*03:01 MHC complex; (iii) a peptide conjugate comprising a peptide consisting of the amino acid sequence KLVVVGACGV / HLA-A*02:01 MHC complex; or any combination of (i)-(iii) (e.g., (i) and (ii); or (i) and (iii)); or all of (i)-(iii). Covalent inhibitors targeting peptide conjugate / MHC complexes may have a chemical structure comprising C-R1, where C is a chemical moiety attached to R1 of any of the compounds shown below.

[0297]

[0298] The R1 of the C-R1 chemical structure of the covalent inhibitor can be any of the structures shown below.

[0299]

[0300] Targeted covalent inhibitors with a structure comprising C-RI can form covalent bonds with multiple different amino acid residues on a peptide. For example, a targeted covalent inhibitor can form a covalent bond with a 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 with an aspartic acid residue, a serine residue or an 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 a structure comprising C-R1 of a peptide conjugate / MHC complex. In certain other embodiments, the antigen-binding domain of the polypeptide recognizes the C portion of the targeted covalent inhibitor of the peptide conjugate / MHC complex but does not recognize the R1 portion.

[0301] Small molecules with electrophilic warhead groups (e.g., targeted covalent inhibitors) can covalently react with cysteine ​​residues of peptides to form peptide-small molecule conjugates. This type of reaction is illustrated in the following scheme for AMG-510 (sotolacibin).

[0302]

[0303] The following table lists the chemical structures of targeted covalent inhibitors and fragments that bind to cysteine ​​residues of peptides after a covalent reaction of the drug with the cysteine ​​("chemical fragments"). In one embodiment, the binding partners disclosed herein bind to a peptide conjugate / MHC complex comprising a peptide conjugated to a chemical fragment listed in Table K below and MHC.

[0304] Table K. Structures of targeted covalent inhibitors and chemical fragments after covalent reaction with cysteine ​​residues.

[0305]

[0306]

[0307]

[0308] Table R. Structures of exemplary targeted covalent inhibitors targeting G12S

[0309]

[0310]

[0311] In one non-limiting embodiment, the binding partner specifically binds to a site comprising a neoantigen comprising a covalently linked small molecule drug or other covalently linked molecule as a component of the antigen in a specific MHC context. In one 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 one embodiment, the binding affinity of the binding partner to the peptide conjugate / MHC complex is greater than the binding affinity to the peptide or free targeted covalent inhibitor.

[0312] In one 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 peptide or free targeted covalent inhibitor. In one 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, or at least 10,000 times greater than the affinity of the binding partner for the free targeted covalent inhibitor or free peptide conjugate. In some embodiments, the affinity of the binding partner for the free drug is reported as an IC 50 In some embodiments, the affinity of the binding partner for the free drug is reported as an IC greater than 1 μM. 50 In some embodiments, the affinity of the binding partner for the free drug is reported as an IC greater than 5 μM. 50 In some embodiments, the affinity of the binding partner for the free drug is reported as an IC greater than 10 μM. 50 In some embodiments, the affinity of the binding partner for the free drug is reported as an IC greater than 15 μM. 50 In some embodiments, the affinity of the binding partner for the free drug is reported as an IC greater than 20 μM. 50 In some embodiments, the affinity of the binding partner for the free drug is reported as an IC greater than 30 μM. 50 In some embodiments, the affinity of the binding partner for the free drug is reported as an IC greater than 40 μM. 50 In some embodiments, the affinity of the binding partner for the free drug is reported as an IC greater than 50 μM. 50 In some embodiments, the affinity of the binding partner for the free drug is reported as an IC greater than 75 μM. 50 In some embodiments, the affinity of the binding partner for the free drug is reported as an IC greater than 100 μM. 50 In some embodiments, the affinity of the binding partner for the free drug is reported as an IC greater than 200 μM.50 .

[0313] In some embodiments, the antigen binding domain does not detectably bind to a free targeted covalent inhibitor. In some embodiments, the antigen binding domain binds to a free targeted covalent inhibitor with an IC of at least about 50 nM. 50 In some embodiments, the antigen binding domain does not bind to the free targeted covalent inhibitor with an IC of less than about 50 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 5 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 25 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 40 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 60 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 75 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 100 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 250 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 500 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 750 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 1000 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 2 μM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 5 μM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 10 μM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 15 μM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 20 μM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 25 μM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of at least about 50 μM. 50 Binds to free targeted covalent inhibitors.

[0314] In some embodiments, the antigen binding domain can be expressed with an IC of less than about 50 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of less than about 5 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of less than about 25 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of less than about 40 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of less than about 60 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of less than about 75 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of less than about 100 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of less than about 250 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of less than about 500 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of less than about 750 nM. 50 The antigen binding domain can bind to a free targeted covalent inhibitor with an IC of less than about 1000 nM. 50 The antigen binding domain can bind to a free target covalent inhibitor with an IC of less than about 2 μM. 50 The antigen binding domain can bind to a free target covalent inhibitor with an IC of less than about 5 μM. 50 The antigen binding domain can bind to a free target covalent inhibitor with an IC of less than about 10 μM. 50 The antigen binding domain can bind to a free target covalent inhibitor with an IC of less than about 15 μM. 50 The antigen binding domain can bind to a free target covalent inhibitor with an IC of less than about 20 μM. 50 The antigen binding domain can bind to a free target covalent inhibitor with an IC of less than about 25 μM. 50 The antigen binding domain can bind to a free target covalent inhibitor with an IC of less than about 50 μM. 50 Binds to free targeted covalent inhibitors.

[0315] The binding partners described herein may not bind to the peptide / MHC complex without a conjugate (e.g., a targeted covalent inhibitor). In some cases, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as a dissociation constant (K) of 10 pM to 50 nM. DIn some cases, the affinity of the binding partner to the peptide conjugate / MHC complex is reported as a dissociation constant (K) of 1 pM to 10 pM. D In some cases, the affinity of the binding partner to the peptide conjugate / MHC complex is reported as a dissociation constant (K) of 0.01 pM to 10 pM. D In some cases, the affinity of the binding partner to the peptide conjugate / MHC complex is reported as a dissociation constant (K) of 0.1 nM to 1 nM. D In some cases, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as a dissociation constant (K) ranging from 1 nM to 2 nM. D In some cases, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as a dissociation constant (K) ranging from 2 nM to 5 nM. D In some cases, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as a dissociation constant (K) equal to or less than about 5 nM. D In some cases, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as a dissociation constant (K) equal to or less than about 4 nM. D In some cases, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as a dissociation constant (K) equal to or less than about 3 nM. D In some cases, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as a dissociation constant (K) equal to or less than about 2 nM. D In some cases, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as a dissociation constant (K) equal to or less than about 1 nM. D In some cases, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as a dissociation constant (K) equal to or less than about 0.1 nM. D In some cases, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as a dissociation constant (K) of approximately equal to or less than 0.01 nM. D In some cases, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as a dissociation constant (K) equal to or less than about 1 pM. D In some cases, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as a dissociation constant (K) equal to or less than about 0.1 pM. D ).

[0316] In some embodiments, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as IC 50 or EC 50In some embodiments, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as an IC of less than about 1 pM. 50 In some embodiments, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as an IC of less than about 10 pM. 50 In some embodiments, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as an IC of less than about 100 pm 50 In some embodiments, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as an IC of less than about 1000 pm 50 In some embodiments, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as an IC50 of less than about 1 nM. In some embodiments, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as an IC50 of less than about 10 nM. 50 In some embodiments, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as an IC of less than about 100 nM. 50 In some embodiments, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as an IC of less than about 1000 nM. 50 In some embodiments, the affinity of the binding partner for the peptide conjugate / MHC complex is reported as an IC of less than about 1 μM. 50 .

[0317] In some embodiments, the binding partner does not detectably bind to a complex of a peptide and an MHC molecule, wherein the peptide is not covalently bound to non-peptide molecules. In some embodiments, the binding partner binds to a complex of a peptide and an MHC molecule with an affinity of less than 10x, wherein the peptide is not covalently bound to non-peptide molecules. In some embodiments, the binding partner binds to a complex of a peptide and an MHC molecule with an affinity of less than 100x, wherein the peptide is not covalently bound to non-peptide molecules. In some embodiments, the binding partner binds to a complex of a peptide and an MHC molecule with an affinity of less than 1,000x, wherein the peptide is not covalently bound to non-peptide molecules.

[0318] In some embodiments, the binding partner can be a polypeptide. In some embodiments, the polypeptide can have an antigen binding domain. The dissociation constant (K) of the antigen binding domain binding to the peptide conjugate / MHC complex D ) can be 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, or at most about 1 pM. The dissociation constant (K) for binding of the antigen binding domain to the peptide conjugate / MHC complex is D ) can be from about 0.01 nM to about 20 nM.

[0319] The antigen binding domain can bind to the free target covalent inhibitor with a dissociation constant (K D ) is higher than the K of the antibody or antigen-binding fragment and peptide conjugate / MHC complex D In some cases, the antigen binding domain can bind to a free targeted covalent inhibitor with a dissociation constant (K D ) is at least about 100 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 μM, at least about 10 μM, at least about 20 μM, at least about 30 μM, at least about 40 μM, at least about 50 μM, or at least 100 μM. The antigen binding domain can bind to a free targeted covalent inhibitor with a dissociation constant (K D ) is the K of the antigen binding domain binding to the peptide conjugate / MHC complex D The antigen binding domain can bind to the free peptide conjugate with a dissociation constant (K D ) is higher than the K of the antibody or antigen-binding fragment and peptide conjugate / MHC complex D The antigen binding domain can bind to the free peptide conjugate with a dissociation constant (K D ) is greater than about 100 nM, greater than about 200 nM, greater than about 300 nM, greater than about 400 nM, greater than about 500 nM, greater than 1 μM, greater than 10 μM, greater than 20 μM, greater than 30 μM, greater than 40 μM, greater than 50 μM or greater than 100 μM. The antigen binding domain can bind to the free peptide conjugate with a dissociation constant (K D ) is the K of the antigen binding domain binding to the peptide conjugate / MHC complex D The antigen binding domain can bind to the free peptide conjugate with a dissociation constant (K D ) is the K of the antibody or antigen-binding fragment and peptide conjugate / MHC complex Dof at least 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5. In some embodiments, the dissociation constant (K) of the antigen binding domain for binding to the free peptide conjugate is 5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, or 10. D ) is the K of the antibody or antigen-binding fragment and peptide conjugate / MHC complex D The antigen binding domain can bind to the peptide conjugate / MHC complex with an affinity that is at least 100-fold, at least 200-fold, at least 300-fold, at least 400-fold, at least 500-fold, at least 600-fold, at least 700-fold, at least 800-fold, at least 900-fold, at least 1,000-fold, at least 2,500-fold, at least 5,000-fold, or at least 10,000-fold greater than the affinity of the antigen binding domain to the free targeted covalent inhibitor or the free peptide conjugate. The MHC can be HLA-A02:01. The MHC can be HLA-A03:01.

[0320] In one embodiment, the MHC is a human leukocyte antigen (HLA). In one embodiment, the HLA is any HLA found in the allele frequency network database (allelefrequencies.net). In one embodiment, the HLA is HLA-A*02:01, HLA-A*03:01, HLA-A*01:01, HLA-A*02:01, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*26:01, HLA-B*07:02, HLA-B*08:01, HLA-B*27:05, HLA-B*39:01, HLA-B*40:01, HLA-B*58:01, HLA-B*15:01 and / or HLA-A*11:01.

[0321] In one aspect, a cell-free peptide conjugate / MHC complex is provided herein, comprising: (a) a peptide conjugate formed by a covalent reaction of a targeted covalent inhibitor with a peptide; and (b) MHC. In some embodiments, the peptide comprises a fragment of RAS (e.g., KRAS G12C 、KRAS G12D 、KRAS G12R 、KRAS G12S , HRAS G12C , HRAS G12D , HRAS G12R , HRAS G12S NRAS G12C NRAS G12D NRAS G12R or NRAS G12S ), EGFR, BTK, HER2 / NEU (ERBB2), HER3 (ERBB3), HER4 (ERBB4), MET (HGFR); FGFR, CDK, acetylcholinesterase (ACHE), p90 ribosomal S6 kinase (RSK), TP53, IDH1, GNAS, FBXW7, CTNNB1, DNMT3A, cathepsin B, cathepsin C, cathepsin F, cathepsin H, cathepsin K, cathepsin L, cathepsin O, cathepsin S, cathepsin V, cathepsin W, cathepsin X, caspase, pancreatic lipase, METAP2, any cancer testis antigen, any endogenous retroviral protein, long interspersed element-1 (LINE-1) or short interspersed element (SINE). In one embodiment, the short interspersed element is Alu. In one embodiment, the targeted covalent inhibitor is osimertinib, ibrutinib, neratinib, AMG-510, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1, or BI 182391. Examples of additional compounds that can covalently target KRAS peptides comprising the G12C mutation can be found in International Application No. PCT / IB2019 / 050993, International 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 other embodiments, the targeted covalent inhibitor is selected from the group consisting of β-lactone G12Si-1, G12Si-2, G12Si-3, G12Si-4, and G12Si-5.

[0322] In one aspect, provided herein is a cell-free peptide conjugate / MHC complex comprising: (a) a compound selected from the group consisting of compounds 1 and 4-8,

[0323]

[0324]

[0325] It covalently binds to a cysteine ​​residue in a peptide comprising the amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV; and (b) MHC.

[0326] In one aspect, provided herein is a cell-free peptide conjugate / MHC complex comprising: (a) Compound 2:

[0327]

[0328] It covalently binds to a cysteine ​​residue in a peptide comprising the amino acid sequence of QLMPFGCLL, LMPFGCLLDY or MPFGCLLDY; and (b) MHC.

[0329] In one aspect, provided herein is a cell-free peptide conjugate / MHC complex comprising: (a) Compound 3:

[0330]

[0331] It covalently binds to a cysteine ​​residue in a peptide comprising the amino acid sequence of YMANGCLLNY; and (b) MHC.

[0332] In one embodiment, the MHC is HLA, optionally wherein the HLA is HLA-A*02:01, HLA-A*03:01, HLA-A*01:01, HLA-A*02:01, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*26:01, HLA-B*07:02, HLA-B*08:01, HLA-B*27:05, HLA-B*39:01, HLA-B*40:01, HLA-B*58:01, HLA-B*15:01 or HLA-A*11:01.

[0333] In one aspect, provided herein is a binding partner that specifically binds to KRAS presented by AMG-510 and HLA. G12C The conjugate is formed by the covalent reaction of a peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein

[0334] (a) VH comprises:

[0335] (i) CDR-H1 comprising the amino acid sequence of DYSIH, or a variant thereof comprising 1-5 amino acid changes;

[0336] (ii) CDR-H2 comprising the amino acid sequence of SISSSSGSTSYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and / or

[0337] (iii) CDR-H3 comprising an amino acid sequence of GX1WX2X3AMDY, wherein X1 is G, R, H, S or K, X2 is Y or I, and X3 is P or A; and / or

[0338] (b) VL contains:

[0339] (i) CDR-L1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;

[0340] (ii) CDR-L2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes; and / or

[0341] (iii) CDR-L3 comprising an amino acid sequence of QQX1SYVX2X3X4IT, wherein X1 is I, A, P, V or S, X2 is K, R, A or H, X3 is K or R, and X4 is L, T, K, R, V, A or E.

[0342] The CDR sequences of exemplary antibodies that bind to conjugates formed by the covalent reaction of AMG-510 with KRAS(G12C) peptide presented on HLA-A*03:01 and HLA-A*11:01 are provided in Tables G and H below.

[0343] Table G: Heavy chain CDR sequences of exemplary antibodies that bind to conjugates formed by covalent reaction of AMG-510 with KRAS(G12C) peptide presented on HLA-A*03:01 and HLA-A*11:01.

[0344]

[0345]

[0346] Table H: Light chain CDR sequences of exemplary antibodies that bind to conjugates formed by covalent reaction of AMG-510 with KRAS(G12C) peptide presented on HLA-A*03:01 and HLA-A*11:01.

[0347]

[0348] In one embodiment, the binding partner comprises CDR-H1, CDR-H2 and CDR-H3 of the heavy chain variable region and / or CDR-L1, CDR-L2 and CDR-L3 of the light chain variable region of a binding partner selected from the group consisting of: RA_D11, RA_D01-RA_D04, RA_D06-RA_D09, RA_D12-RA_D14, RA_D16, RA_D18-RA_D21, RA_D23 and RA_D24, or a variant thereof comprising 1-5 amino acid changes in one or more CDR amino acid sequences.

[0349] In one embodiment, the binding partner comprises a CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and / or CDR-L3 amino acid sequence of a binding partner selected from the group consisting of RA_D11, RA_D01-RA_D04, RA_D06-RA_D09, RA_D12-RA_D14, RA_D16, RA_D18-RA_D21, RA_D23 and RA_D24 (Table G and Table H), or a variant thereof comprising 1-5 amino acid changes in one or more CDR amino acid sequences.

[0350] In one embodiment, the binding partner may comprise a VH and / or 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.

[0351] In one embodiment, the binding partner may comprise a VH and / or 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 VH and / or VL of RA_D11, RA_D01-RA_D04, RA_D06-RA_D09, RA_D12-RA_D14, RA_D16, RA_D18-RA_D21, RA_D23 and RA_D24 (Table G and Table H).

[0352] AMG-510 binds to KRAS G12C A non-limiting example of a reference binding partner for a conjugate formed by covalent reaction of peptides and presented by an HLA molecule is referred to herein as RA_D11.

[0353] The light chain variable region (V L ) and heavy chain variable region (VH ) is (CDRs are shown in bold; underlining corresponds to residues shown in Tables E and F, which are given in the figures):

[0354]

[0355] A non-limiting example of a reference binding partner that binds to KRAS(G12C)-AMG-510 presented by an HLA molecule is referred to herein as AMRA3-7D. The light chain variable region (V L ) and heavy chain variable region (V H ) has an amino acid sequence of:

[0356]

[0357] Figure 25 Shown are data including a comparison of binding of AMRA3-7D and RA-D11 (and other binding partners) to the AMG510-G12C 9mer in an HLA-A*11 background.

[0358] In another aspect, the present disclosure provides a binding partner (or antigen binding domain) with improved binding affinity for a drug-peptide / MHC complex. The peptide can be a KRAS peptide. The drug can be sotolacib (e.g., AMG-510). The drug-peptide conjugate can be a KRAS (G12C) peptide conjugated to sotolacib. The binding partner can bind to KRAS (G12C)-sotolacib presented by an MHC molecule. In certain aspects, the MHC can be HLA-A*02:01, HIA-A*03:01, or HLA-A*11:01. The binding partners described herein may comprise various antigen binding domains disclosed herein.

[0359] For example, in some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), wherein VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GSWIHAMDY. VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SISSSWGVTSYADSVKG. VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FHWYSIH. In some embodiments, the antigen-binding domain may further comprise a light chain variable region (VL), wherein VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. VL may further comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. VL may further comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise a CDR-H3 sequence of GSWIHAMDY; a CDR-H2 sequence of SISSSWGVTSYADSVKG; a CDR-H1 sequence of FHWYSIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFHWYSIHWVRQAPGKGLEWVASISSSWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIHAMDYWCQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to: EVQLVESGGGLVQPGGSLRLSCAASGFTFHWYSIHWVRQAPGKGLEWVASISSSWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIHAMDYWGQGTLVTVSS.The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV. The VL may comprise a sequence having at least 80% sequence identity to DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0360] In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), wherein VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GHWIAAMDY. VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SIASSSGSTGYADSVKG. VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FSWYSIH. In some embodiments, the antigen binding domain may further comprise a light chain variable region (VL), wherein VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. VL may further comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. VL may further comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise a CDR-H3 sequence of GHWIAAMDY; a CDR-H2 sequence of SIASSSGSTGYADSVKG; a CDR-H1 sequence of FSWYSIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSWYSIHWVRQAPGKGLEWVASIASSSGSTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGHWIAAMDYWGQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSWYSIHWVRQAPGKGLEWVASIASSSGSTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGHWIAAMDYWGQGTLVTVSS. The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the following sequence: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.The VL may comprise a sequence having at least 80% sequence identity to: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0361] In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), wherein VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GGVIHAMDY. VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SILSRWGVTSYADSVKG. VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FSPYSIH. In some embodiments, the antigen binding domain may further comprise a light chain variable region (VL), wherein VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. VL may further comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. VL may further comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise a CDR-H3 sequence of GGVIHAMDY; a CDR-H2 sequence of SILSRWGVTSYADSVKG; a CDR-H1 sequence of FSPYSIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYSIHWVRQAPGKGLEWVASILSRWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGVIHAMDYWGQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYSIHWVRQAPGKGLEWVASILSRWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGVIHAMDYWGQGTLVTVSS. The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the following sequence: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.The VL may comprise a sequence having at least 80% sequence identity to: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0362] In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), wherein the VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GSWIAAMDY. The VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SISSWHGETGYADSVKG. The VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FSPYSIH. In some embodiments, the antigen-binding domain may further comprise a light chain variable region (VL), wherein the VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. The VL may comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. The VL may comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise: a CDR-H3 sequence of GSWIAAMDY; a CDR-H2 sequence of SISSWHGETGYADSVKG; a CDR-H1 sequence of FSPYSIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYSIHWVRQAPGKGLEWVASISSWHGETGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYSIHWVRQAPGKGLEWVASISSWHGETGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS. The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the following sequence: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.The VL may comprise a sequence having at least 80% sequence identity to: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0363] In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), wherein the VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GGWIAAMDY. The VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SISSLQGDTGYADSVKG. The VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FSWYSIH. In some embodiments, the antigen-binding domain may further comprise a light chain variable region (VL), wherein the VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. The VL may further comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. The VL may further comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise a CDR-H3 sequence of GGWIAAMDY; a CDR-H2 sequence of SISSLQGDTGYADSVKG; a CDR-H1 sequence of FSWYSIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSWYSIHWVRQAPGKGLEWVASISSLQGDTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGWIAAMDYWGQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSWYSIHWVRQAPGKGLEWVASISSLQGDTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGWIAAMDYWGQGTLVTVSS. The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the following sequence: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.The VL may comprise a sequence having at least 80% sequence identity to: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0364] In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), wherein VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GSWIAAMDY. VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SIASWYGDTGYADSVKG. VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FHYYSIH. In some embodiments, the antigen-binding domain may also comprise a light chain variable region (YL), wherein VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. VL may comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. VL may comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise: a CDR-H3 sequence of GSWIAAMDY; a CDR-H2 sequence of SIASWYGDTGYADSVKG; a CDR-H1 sequence of FHYYSIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFHYYSIHWVRQAPGKGLEWVASIASWYGDTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFHYYSIHWVRQAPGKGLEWVASIASWYGDTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS. The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the following sequence: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.The VL may comprise a sequence having at least 80% sequence identity to: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0365] In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), wherein VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GGRIEAMDY. VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SISSWYGKTGYADSVKG. VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FGYYSIH. In some embodiments, the antigen-binding domain may further comprise a light chain variable region (VL), wherein VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. VL may further comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. VL may further comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise a CDR-H3 sequence of GGRIEAMDY; a CDR-H2 sequence of SISSWYGKTGYADSVKG; a CDR-H1 sequence of FGYYSIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFGYYSIHWVRQAPGKGLEWVASISSWYGKTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGRIEAMDYWGQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFGYYSIHWVRQAPGKGLEWVASISSWYGKTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGRIEAMDYWGQGTLVTVSS. The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the following sequence: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.The VL may comprise a sequence having at least 80% sequence identity to: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0366] In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), wherein VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GYWIEAMDY. VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SIASSYGSTGYADSVKG. VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FSKYSIH. In some embodiments, the antigen-binding domain may also comprise a light chain variable region (VI), wherein VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. VL may comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. VL may comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise: a CDR-H3 sequence of GYWIEAMDY; a CDR-H2 sequence of SIASSYGSTGYADSVKG; a CDR-H1 sequence of FSKYSIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSKYSIHWVRQAPGKGLEWVASIASSYGSTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYWIEAMDYWGQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSKYSIHWVRQAPGKGLEWVASIASSYGSTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYWIEAMDYWGQGTLVTVSS. The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the following sequence: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.The VL may comprise a sequence having at least 80% sequence identity to: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0367] In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), wherein VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GSWIAAMDY. VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SIHSSIGTTGYADSVKG. VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FGLYSIH. In some embodiments, the antigen-binding domain may also comprise a light chain variable region (VL), wherein VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. VL may comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. VL may comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise: a CDR-H3 sequence of GSWIAAMDY; a CDR-H2 sequence of SIHSSIGTTGYADSVKG; a CDR-H1 sequence of FGLYSIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFGLYSIHWVRQAPGKGLEWVASIHSSIGTTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFGLYSIHWVRQAPGKGLEWVASIHSSIGTTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS. The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the following sequence: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.The VL may comprise a sequence having at least 80% sequence identity to: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0368] In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), wherein VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GSVIHAMDY. VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SILSWIGKTSYADSVKG. VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FSPYSIH. In some embodiments, the antigen binding domain may further comprise a light chain variable region (VL), wherein VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. VL may further comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. VL may further comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise: a CDR-H3 sequence of GSVIHAMDY; a CDR-H2 sequence of SILSWIGKTSYADSVKG; a CDR-H1 sequence of FSPYSIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYSIHWVRQAPGKGLEWVASILSWIGKTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSVIHAMDYWGQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYSIHWVRQAPGKGLEWVASILSWIGKTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSVIHAMDYWGQGTLVTVSS. The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the following sequence: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.The VL may comprise a sequence having at least 80% sequence identity to: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0369] In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), wherein VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GGWIAAMDY. VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SIASRWGHTGYADSVKG. VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FSPYHIH. In some embodiments, the antigen-binding domain may also comprise a light chain variable region (VL), wherein VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. VL may comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. VL may comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise a CDR-H3 sequence of GGWIAAMDY; a CDR-H2 sequence of SIASRWGHTGYADSVKG; a CDR-H1 sequence of FSPYHIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYHIHWVRQAPGKGLEWVASIASRWGHTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGWIAAMDYWGQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYHIHWVRQAPGKGLEWVASIASRWGHTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGWIAAMDYWGQGTLVTVSS. The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the following sequence: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.The VL may comprise a sequence having at least 80% sequence identity to: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0370] In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), wherein VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GSWIAAMDY. VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SIASLQGITGYADSVKG. VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FHEYSIH. In some embodiments, the antigen-binding domain may also comprise a light chain variable region (VL), wherein VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. VL may comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. VL may comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise: a CDR-H3 sequence of GSWIAAMDY; a CDR-H2 sequence of SIASLQGITGYADSVKG; a CDR-H1 sequence of FHEYSIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFHEYSIHWVRQAPGKGLEWVASIASLQGITGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFHEYSIHWVRQAPGKGLEWVASIASLQGITGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS. The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the following sequence: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.The VL may comprise a sequence having at least 80% sequence identity to: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0371] In some embodiments, the antigen binding domain comprises a heavy chain variable region (VH), wherein VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GGVIHAMDY. VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SILSRWGVTSYADSVKG. VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FSDYSIH. In some embodiments, the antigen binding domain may further comprise a light chain variable region (VL), wherein VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. VL may further comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. VL may further comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise a CDR-H3 sequence of GGVIHAMDY; a CDR-H2 sequence of SILSRWGVTSYADSVKG; a CDR-H1 sequence of FSDYSIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYSIHWVRQAPGKGLEWVASILSRWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGVIHAMDYWGQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYSIHWVRQAPGKGLEWVASILSRWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGVIHAMDYWGQGTLVTVSS. The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the following sequence: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.The VL may comprise a sequence having at least 80% sequence identity to: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0372] In some embodiments, the antigen-binding domain comprises a heavy chain variable region (VH), wherein VH may comprise a heavy chain complementary determining region 3 (CDR-H3), wherein the CDR-H3 comprises the amino acid sequence of GGVIHAMDY. VH may further comprise a CDR-H2, wherein the CDR-H2 comprises the amino acid sequence of SISSRWGVTSYADSVKG. VH may further comprise a CDR-H1, wherein the CDR-H1 comprises the amino acid sequence of FSDYSIH. In some embodiments, the antigen-binding domain may further comprise a light chain variable region (VL), wherein VL may comprise a CDR-L3, wherein the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT. VL may further comprise a CDR-L2, wherein the CDR-L2 comprises the amino acid sequence of SASSLYS. VL may further comprise a CDR-L1, wherein the CDR-L1 comprises the amino acid sequence of RASQSVSSAVA. The antigen binding domain may comprise a CDR-H3 sequence of GGVIHAMDY; a CDR-H2 sequence of SISSRWGVTSYADSVKG; a CDR-H1 sequence of FSDYSIH; a CDR-L3 sequence of QQASYVRKTIT; a CDR-L2 sequence of SASSLYS; and a CDR-L1 sequence of RASQSVSSAVA. The VH may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYSIHWVRQAPGKGLEWVASISSRWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGVIHAMDYWGQGTLVTVSS. The VH may comprise a sequence having at least 80% sequence identity to the following sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYSIHWVRQAPGKGLEWVASISSRWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGVIHAMDYWGQGTLVTVSS. The VL may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the following sequence: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.The VL may comprise a sequence having at least 80% sequence identity to: DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

[0373] The CDRs of any binding partner or antigen-binding domain described herein can be designated by the Kabat numbering scheme. In some cases, light chain (LC) CDRs can be designated by the Kabat numbering scheme. In some cases, LC CDRs can be designated by a modified Kabat numbering scheme. In some cases, heavy chain (HC) CDRs can be designated by the Kabat numbering scheme. In some cases, HC CDRs can be designated by a modified Kabat numbering scheme. For example, a CDR can contain one or more additional amino acids than a CDR designated by the Kabat numbering scheme.

[0374] In one aspect, provided herein is a binding partner that specifically binds to a conjugate formed by the covalent reaction of osimertinib and an HLA-presented EGFR peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein

[0375] (a) VH comprises:

[0376] (i) CDR-H1 comprising the amino acid sequence of SSYIH, or a variant thereof comprising 1-5 amino acid changes;

[0377] (ii) CDR-H2 comprising the amino acid sequence of YISPSYGSTSYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and / or

[0378] (iii) CDR-H3 comprising an amino acid sequence of EX1X2X3MX4X5DY, wherein X1 is Y, L, S or E, X2 is V, T or I, X3 is T or I, X4 is A, T or S, and X5 is L, A, I, K, P or T; and / or

[0379] (b) VL contains:

[0380] (i) CDR-L1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes;

[0381] (ii) CDR-L2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes; and / or

[0382] (iii) CDR-L3, comprising an amino acid sequence of QQYX1X2WPX3T, wherein X1 is S or A, or S; X2 is Y, H, A, D, E, K, S or G; and X3 is I or E.

[0383] Tables I and J below provide CDR sequences of exemplary antibodies that bind to conjugates formed by the covalent reaction of osimertinib with an EGFR peptide presented on HLA.

[0384] Table I: Heavy chain CDR sequences of exemplary antibodies that bind to conjugates formed by the covalent reaction of osimertinib with EGFR peptide presented on HLA.

[0385] Antibody CDRH1 CDRH2 CDRH3 OEA2-5 SSYIH YISPSYGSTSYADSVKG EYVTMALDY EO_Q01 SSYIH YISPSYGSTSYADSVKG ELVTMTADY EO_Q02 SSYIH YISPSYGSTSYADSVKG EYTTMSIDY EO_Q03 SSYIH YISPSYGSTSYADSVKG ESVTMSADY EO_Q04 SSYIH YISPSYGSTSYADSVKG ESVTMTKDY EO_Q05 SSYIH YISPSYGSTSYADSVKG EEVTMSIDY EO_Q06 SSYIH YISPSYGSTSYADSVKG ESITMTKDY EO_Q07 SSYIH YISPSYGSTSYADSVKG ELTTMSIDY EO_Q08 SSYIH YISPSYGSTSYADSVKG ELTEMTTDY EO_Q09 SSYIH YISPSYGSTSYADSVKG EEVTMTADY EO_Q10 SSYIH YISPSYGSTSYADSVKG ELIEMTPDY EO_Q11 SSYIH YISPSYGSTSYADSVKG ELVTMTIDY EO_Q12 SSYIH YISPSYGSTSYADSVKG ELTTMSIDY EO_Q13 SSYIH YISPSYGSTSYADSVKG ELTTMTADY EO_Q14 SSYIH YISPSYGSTSYADSVKG ESITMSPDY EO_Q15 SSYIH YISPSYGSTSYADSVKG ELIEMTTDY EO_Q16 SSYIH YISPSYGSTSYADSVKG EYVTMSIDY EO_Q17 SSYIH YISPSYGSTSYADSVKG ELVEMTPDY EO_Q18 SSYIH YISPSYGSTSYADSVKG EYVTMALDY EO_Q20 SSYIH YISPSYGSTSYADSVKG EEVTMTPDY EO_Q22 SSYIH YISPSYGSTSYADSVKG EYVTMALDY EO_Q23 SSYIH YISPSYGSTSYADSVKG EYVTMALDY EO_Q24 SSYIH YISPSYGSTSYADSVKG EYVTMALDY

[0386] Table J: Light chain CDR sequences of exemplary antibodies that bind to the conjugate formed by the covalent reaction of osimertinib with the EGFR peptide presented on HLA.

[0387] Antibody CDR-L1 CDR-L2 CDR-L3 OEA2-5 RASQSVSSAVA SASSLYS QQYSYWPIT EO_Q01 RASQSVSSAVA SASSLYS QQYSHWPIT EO_Q02 RASQSVSSAVA SASSLYS QQYSAWPET EO_Q03 RASQSVSSAVA SASSLYS QQYSAWPIT EO_Q04 RASQSVSSAVA SASSLYS QQYSDWPIT EO_Q05 RASQSVSSAVA SASSLYS QQYAEWPIT EO_Q06 RASQSVSSAVA SASSLYS QQYSHWPIT EO_Q07 RASQSVSSAVA SASSLYS QQYAKWPIT EO_Q08 RASQSVSSAVA SASSLYS QQYSSWPIT EO_Q09 RASQSVSSAVA SASSLYS QQYSHWPIT EO_Q10 RASQSVSSAVA SASSLYS QQYSGWPIT EO_Q11 RASQSVSSAVA SASSLYS QQYSGWPIT EO_Q12 RASQSVSSAVA SASSLYS QQYASWPIT EO_Q13 RASQSVSSAVA SASSLYS QQYSHWPIT EO_Q14 RASQSVSSAVA SASSLYS QQYSSWPIT EO_Q15 RASQSVSSAVA SASSLYS QQYSHWPIT EO_Q16 RASQSVSSAVA SASSLYS QQYSDWPIT EO_Q17 RASQSVSSAVA SASSLYS QQYSSWPIT EO_Q18 RASQSVSSAVA SASSLYS QQYSAWPET EO_Q20 RASQSVSSAVA SASSLYS QQYSDWPIT EO_Q22 RASQSVSSAVA SASSLYS QQYSHWPIT EO_Q23 RASQSVSSAVA SASSLYS QQYSHWPET EO_Q24 RASQSVSSAVA SASSLYS QQYSEWPET

[0388] In one embodiment, the binding partner comprises CDR-H1, CDR-H2 and CDR-H3 of the heavy chain variable region amino acid sequence and / or CDR-L1, CDR-L2 and CDR-L3 of the light chain variable region of the binding partner selected from the group consisting of: OEA2-5, EO_Q01-EO_Q18 and EO_Q20-EO_Q24, or a variant thereof comprising 1-5 amino acid changes in one or more CDR amino acid sequences.

[0389] In one embodiment, the binding partner comprises a CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and / or CDR-L3 amino acid sequence of a binding partner selected from the group consisting of: OEA2-5, EO_Q01-EO_Q18 and EO_Q20-EO_Q24 (Tables I and J), or a variant thereof comprising 1-5 amino acid changes in one or more CDR amino acid sequences.

[0390] In one embodiment, the binding partner may comprise a VH and / or 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 VH and / or VL amino acid sequence of an antibody disclosed in Table I and Table J.

[0391] A non-limiting example of a binding partner that binds to the epidermal growth factor receptor (EGFR)-osimertinib conjugate is referred to herein as OEA2-5. The present disclosure includes all derivatives of OEA2-5 described herein, including the substituted residues described below in scDb format by deep mutational analysis, and provides their representative amino acid sequences.

[0392] The light chain variable region (V L ) and heavy chain variable region (V H ) has an amino acid sequence of:

[0393]

[0394] In some embodiments, the binding partner binds to the native form of the protein (except for the drug or other molecule covalently linked thereto). "Native form" refers to a complete protein that retains its biological function before the drug or other molecule is covalently linked. In some embodiments, the native form of a protein is its form before being fragmented, for example, by intracellular processing. In some embodiments, the binding protein is therefore bound to a full-length polypeptide covalently linked to a drug or other molecule, wherein the covalently bound drug or other molecule at least partially allows preferential binding of the binding partner. Generally speaking, a polypeptide (which is used interchangeably with the term "protein" in this article) comprises more than 50 consecutive amino acids. In some embodiments, the binding partner specifically binds to the complete protein covalently linked to a drug or other molecule. In other embodiments, the binding partner specifically binds to a peptide comprising a covalently bound molecule. In some embodiments, the binding partner specifically binds to a peptide having a specific amino acid sequence and is covalently conjugated to another molecule (e.g., a drug). In some embodiments, the binding partner preferentially binds to a peptide covalently bound to a molecule (e.g., a drug), wherein the sequence of the peptide is unrelated. This preferential binding is relative to the binding of the same peptide not conjugated to the drug. In some embodiments, the binding partner preferentially binds to a peptide comprising a KRAS(G12) mutation or 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 variant thereof that is not covalently conjugated to a drug or other molecule.

[0395] In some embodiments, the binding partner specifically binds to a peptide conjugate of appropriate length for presentation in the major histocompatibility complex (MHC), which in humans is known as human leukocyte antigen (HLA), or in non-human animals (including non-human mammals) as MHC or its equivalent complex.

[0396] In general, peptide conjugates contain less than 50 consecutive amino acids. Thus, in some embodiments, the length of the peptide conjugate comprising the epitope may be 2-49 amino acids. In some embodiments, the peptide covalently linked to the drug or other molecule and the one or more residues of the linked drug and peptide may be composed of an epitope comprising 4-12 consecutive amino acids, which may or may not come from a longer protein during protein processing, such as processing for antigens presented by MHC molecules. In some embodiments, the drug is conjugated to a peptide comprising or consisting of 7-30 amino acids. In some embodiments, the drug or other molecule is conjugated to a peptide comprising or consisting of 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids and can be presented in the context of MHC class I. In some embodiments, the drug or other molecule is conjugated to a peptide having 9-30 amino acids (including the end values ​​and all values ​​and ranges therebetween) and can be presented in the context of MHC class II. In some embodiments, the drug or other molecule is conjugated to a peptide comprising at least 7 amino acids. Non-classical MHC class I molecules are used to mediate inhibitory or activating stimulation in natural killer (NK) cells. Non-classical MHC class I molecules can be expressed by immune cells and tumor cells. For example, the expression of non-classical MHC complexes on malignant cells can hinder the 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, drugs or other molecules are conjugated to peptides with 9-30 amino acids (including end values, and including all numerical values ​​and numerical ranges therebetween) and that can be presented in the context of non-classical MHC class I. In some embodiments, peptide conjugates form complexes with non-classical MHC class I molecules. In some embodiments, non-classical MHC class I molecules are selected from the group consisting of HLA-E, HLA-F and HLA-G. In some embodiments, non-classical MHC class I molecules are selected from HLA-E, HLA-F, HLA-G or a certain combination thereof.

[0397] In some embodiments, the binding partner specifically binds to a peptide conjugate covalently conjugated to a drug or other molecule independent of MHC presentation. In some embodiments, non-limiting examples of which are described in Example 3 below, the binding partner specifically binds to the peptide conjugate only when the peptide conjugate is presented by an MHC molecule. In some embodiments, the binding partner can specifically bind to the peptide conjugate in both MHC-independent and MHC-presentation contexts. In some embodiments, the MHC-peptide conjugate complex comprises the antigen to which the binding partner specifically binds.

[0398] In some embodiments, the binding partner exhibits at least one improved property relative to the same property of a reference binding partner. In some embodiments, the reference binding partner is AMRA3-7D. In one embodiment, the binding partner exhibits greater affinity for its target relative to the reference binding partner.

[0399] In some embodiments, the binding partner can thus bind to cells via any MHC capable of presenting the peptide conjugate. In some embodiments, the HLA is expressed by cells capable of class I, class II, or class III MHC presentation. In some embodiments, the binding partner can bind to cells expressing class I MHC that present the peptide conjugate. Those skilled in the art will recognize that class I MHC includes components such as polymorphic α chains and β2 microglobulin, wherein the peptide conjugate binds to the polymorphic chain.

[0400] In some embodiments, the cell is an antigen presenting cell (APC). In some embodiments, the cell is a so-called professional antigen presenting cell, and thus may include but is not limited to macrophages and dendritic cells, which display class II MHC. Those skilled in the art will recognize that class II MHC includes components such as MHC polymorphic α and β chains, and that the displayed peptide conjugate is bound to both chains. In other embodiments, class II MHC may be displayed together with the peptide conjugate by other cell types (e.g., cancer / tumor cells), and thus the present disclosure provides for the use of the binding partner to directly identify such cells without the need for professional APC.

[0401] In some embodiments, the peptide conjugate is displayed by a non-classical MHC complex, which may include CDld, MRl, MHC-E, MHC-F, MHC-G, and / or other emerging family members that will be recognized by those skilled in the art.

[0402] The present disclosure includes binding partners that specifically bind to peptide conjugates displayed only by a particular MHC class, thereby providing binding partners that discriminate between MHC classes. Representative examples of such binding partners are described herein at least by FIG. 17 .

[0403] In some embodiments, the binding partners of the present disclosure can specifically bind to peptide conjugates comprising a covalently conjugated drug or other molecule that are displayed by more than one specific MHC type. In some embodiments, a single binding partner of the present disclosure is applicable to multiple HLA types. In addition, a single binding partner can specifically bind to multiple combinations of (a) peptide conjugates comprising a covalently conjugated drug or other molecule and (b) HLA types, for example, by Figure 35 and Figure 36 In particular, Figure 35 and Figure 36This demonstrates that a single antibody can bind to different peptides conjugated to AMG-510 presented by different HLAs with different peptide binding preferences. For example, comparing the HLA-A*03 / 11 peptides VVVGAC*GVGK and KLVVVGAC*GV, the results demonstrate that antibody RA_D11 still selectively binds to the hapten-peptide-HLA complex relative to binding to the free drug. In some embodiments, the binding partners of the present disclosure can specifically bind to a peptide conjugate comprising a covalently conjugated drug only in a specific MHC context. In some embodiments, the peptide conjugate is displayed by an MHC class I type selected from HLA-A, HLA-B, HLA-C, and combinations thereof. In certain aspects, the peptide conjugate is displayed in any MHC class I context of A*02 / B*35 / C*04. In some embodiments, the peptide conjugate is displayed by any MHC class II of DR*01 / DR*04 / DR*07 / DP*04. In some embodiments, HLA comprises A*01:01, A*02:01, A*03:01, A*11:01, A*24:02, A*26:01, B*07:02, B*08:01, B*27:05, B*39:01, B*40:01, B*58:01, or B*15:01. Specific examples of antibodies include antibodies that bind to KRAS(G12C)-AMG510 conjugates presented on HLA-A*02:01, HLA-A*03:01, and HLA-A*11:01, BTK-ibrutinib conjugates presented on HLA-A*01:01, and EGFR-osimertinib conjugates presented on HLA-A*02:01. In non-limiting embodiments, the present disclosure provides scDbs specific for a particular drug covalently bound to a peptide presented on a particular HLA, or the same drug covalently bound to a peptide presented on two different HLAs. Representative scDbs are described in Example 4. Data obtained using scDbs are presented in Figures 18 and 19. Data obtained using the CrossMa format are provided in Example 5 and its accompanying figures.

[0404] In some embodiments, the present disclosure includes selecting an individual based on the individual's HLA type and selecting an antibody described herein for use in treating the individual. In some embodiments, the selection of a binding partner is based at least in part on the degree of HLA restriction exhibited by the selected binding partner relative to the individual's HLA type.

[0405] In certain embodiments, such as KRAS(G12C) binding partners, representative examples of which bind two different KRAS(G12C)-derived peptides conjugated to a drug (AMG-510, also known as sotolacilib) are provided below.

[0406] In some embodiments, the peptide conjugates are displayed by cells participating in a cell-mediated immune response or by cells that can become targets of a cell-mediated immune response. In some embodiments, the peptide conjugates displayed in any suitable MHC background are presented by cells recognized by leukocytes, including but not necessarily limited to T cells or natural killer (NK) cells. In some embodiments, T cells are CD4+T cells, CD8+T cells, double positive CD4+ / CD8+T cells, CD4+ / CD8+ double negative T cells or γδT cells. Therefore, and as further described below, the present disclosure provides binding partners, which are configured to interact with both the peptide conjugates presented and the cells participating in a cell-mediated immune response. In some embodiments, some of the binding partners are capable of binding to a complex of 1) specific MHC and 2) specific peptide conjugates. In some embodiments, some of the binding partners are capable of binding to a specific peptide conjugate presented by at least two different MHCs.

[0407] In some embodiments, any binding partner of the present disclosure comprises at least one chain, and this chain comprises complementary determining region (CDR), and this complementary determining region (CDR) is CDR1, CDR2 or CDR3 from any heavy chain or light chain amino acid sequence described herein.In certain examples of this specification, CDR is shown in bold.The present disclosure covers the amino acid sequence of CDR sequence respectively by its position in the heavy chain and light chain amino acid sequence.The present disclosure includes the binding partner comprising the heavy chain CDR1, CDR2 and CDR3.The present disclosure also includes the binding partner comprising the light chain CDR1, CDR2 and CDR3.The present disclosure also includes the binding partner comprising the heavy chain CDR1, CDR2 and CDR3 and the light chain CDR1, CDR2 and CDR3.For the amino acid sequence of the present disclosure, it includes the amino acid comprising purification tag or protein production tag (such as HIS tag and / or AVI tag), and the present disclosure includes the following conditions: the sequence of the tag can be excluded from the amino acid sequence. The amino acid between the tags can also be excluded.

[0408] The binding partners of the present disclosure can be provided as intact immunoglobulins or fragments of immunoglobulins, including but not necessarily limited to antigen-binding (Fab) fragments, Fab' fragments, (Fab')2 fragments, Fd (N-terminal portion of the heavy chain) fragments, Fv fragments (two variable domains), diabodies (Db), dAb fragments, single domain fragments or single monomeric variable antibody domains, single-chain diabodies (scDb), isolated complementarity determining regions (CDRs), single-chain variable fragments (scFv) and other antibody fragments that retain antigen binding function. In some embodiments, one or more binding partners are provided as components of a bispecific T cell engaging antibody (BiTE), a bispecific killer cell engaging antibody (BiKE), a CrossMab (e.g., a binding partner comprising four different chains; immunoglobulin crossover (also known as Fab domain exchange or CrossMab format) technology (see, e.g., WO 2009 / 080253; Schaefer et al., Proc. Natl. Acad. Sci. USA, 108: 11187-11192 (2011)), or a chimeric antigen receptor (CAR), e.g., for the generation of chimeric antigen receptor T cells (e.g., CAR T cells) and CAR natural killer (NK) cells, neutrophils, and macrophages. The present disclosure includes binding partners comprising the heavy and light chain variable regions.

[0409] The binding partner of the present disclosure can be a scFv. In the present disclosure, the VH of the polypeptide can be connected to the VL by a linker. The linker can be a peptide linker composed of amino acids such as glycine and / or serine residues used alone or in combination to connect the variable heavy chain region and the variable light chain region 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=1, 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 linker includes but is not limited to (Gly4Ser)4 or (Gly4Ser)3. In another embodiment, the linker includes multiple repeats of (Gly2Ser), (GlySer) or (Gly3Ser). In some cases, the linker sequence includes (G4S) n , wherein n=2 to 4. In some cases, the linker sequence comprises (G4S) n , wherein n=1 to 3. In some cases, the linker may include (G4S) n or (S4G) n , wherein n is any integer between 1 and 10. In some cases, the linker can include a glycine-serine-alanine linker G4SA3 or a glycine-serine linker (G4S)4.

[0410] The binding partner of the present disclosure can be a bispecific T cell engaging antibody (BiTE). BiTE therapy can be used to connect the endogenous T cells of a subject to cancer cells. BiTE molecules can include two Fv fragments of a monoclonal antibody connected by a peptide linker. BiTE molecules can include a first antigen binding domain and a second antigen binding domain. The first antigen binding domain can specifically 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 a cancer cell. In some embodiments, BiTE molecules 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., CD3ε, CD3δ or CD3γ), TCRα chain, TCRβ chain, TCRγ chain, TCRδ chain, ICOS, CD28, CD27, HVEM, LIGHT, CD40, 4-1BB, OX40, DR3, GITR, CD30, TIM1, SLAM, 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 fragment of a protein associated with cancer, optionally wherein the protein is encoded by a gene that mutates in cancer. In some embodiments, the protein is overexpressed in cancer. In some embodiments, a fragment of the 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 includes endogenous retroviruses (ERVs). In some embodiments, the protein is derived from a retrotransposon family, such as a long interspersed element (LINE) or a short interspersed element (SINE). 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 (or colorectal 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 gastric 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).

[0411] The binding partner may 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 partner may comprise two polypeptide chains. For example, the polypeptide may comprise a first polypeptide chain comprising a first antigen-binding domain and a second polypeptide chain comprising a second antigen-binding domain. In some cases, the two polypeptide chains may comprise an Fc region. The binding partner of the present disclosure may be a bispecific or multivalent antibody or antibody fragment 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 may comprise an Fc region of a heavy chain constant region selected from human IgM, IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The Fc region may be a modified version of a wild-type Fc region. For example, the Fc region may be a silent version of the Fc region, including but not limited to LALA Fc, LALAPGFc, LALAKA Fc, LAGA Fc, LGGR Fc, or LALE Fc. The Fc region may comprise one or more mutations of the wild-type Fc region. The Fc region may be connected to one or more tumor targeting moieties, T cell engaging antibodies or cytokine molecules. In some embodiments, the interface of the first Fc region and the second Fc region may be altered to increase or decrease dimerization. The dimerization of the Fc region may be enhanced by providing paired cavity protrusions (e.g., knob-in-hole) to the Fc interface of the first Fc region and the second Fc region. The knob-in-hole described in US5,731,116, US7,476,724 and Ridgway, J. et al. (1996) Prot.Engineering 9(7):617-621 generally involves: (1) mutating the CH3 domains of one or two antibodies to promote heterodimerization; and (2) combining the mutated antibodies under conditions that promote heterodimerization. "Knobs" or "protrusions" can be generated by replacing small amino acids (e.g., T366Y or T366W) in the parent antibody with larger amino acids. " Nest " or " hole " is produced by replacing the larger residue (such as Y407T, T366S, L368A and / or Y407V) in the parent antibody with smaller amino acids. Exemplary ball-and-socket mutations include S354C or T366W in the " ball " heavy chain and Y349C, T366S, L368A or Y407V in the " nest " heavy chain. For bispecific antibodies including Fc domains, specific mutations can be introduced into the constant region of the heavy chain to promote the correct heterodimerization of the Fc portion. These techniques include the ball-and-socket method, which involves introducing a large residue into one of the CH3 domains of one of the antibody heavy chains. The large residue can be inserted into the complementary " nest " of another CH3 domain of the pairing heavy chain, thereby promoting the correct pairing of the heavy chain (see, e.g., US7642228).In some cases, two polypeptide chains can be combined using the ball-and-socket method as described herein. In some cases, two polypeptide chains cannot be combined using the ball-and-socket method. In some cases, the first antigen-binding domain and the second antigen-binding domain are connected by a joint. The joint can be a peptide joint composed of amino acids (e.g., glycine and / or serine residues) used alone or in combination. The length of the joint can include 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 some cases, the joint can include (G4S). n , wherein n is any integer between 1 and 10. In one embodiment, the flexible polypeptide linker is a Gly / Ser linker and comprises the amino acid sequence (Gly-Gly-Gly-Ser)n, wherein n is a positive integer equal to or greater than 1. For example, n=1, 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 linker includes but is not limited to (Gly4Ser)4 or (Gly4Ser)3. In another embodiment, the linker includes multiple repeats of (Gly2Ser), (GlySer) or (Gly3Ser). 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=1 to 3. In some cases, the linker may comprise (G4S) n or (S4G) n , wherein n is any integer between 1 and 10. In some cases, the linker may comprise a glycine-serine-alanine linker G4SA3 or a glycine-serine linker (G4S)4.

[0412] In some embodiments, the cytokine is an engineered cytokine. For example, the engineered cytokine can be a variant or mutant of any cytokine as described herein. The engineered cytokine can be a mutant protein, a super factor or a cytokine with pegylation. In some embodiments, the cytokine is a single-chain cytokine. In some embodiments, the cytokine is a multi-chain cytokine. In some cases, the two polypeptides may comprise two parts. In some cases, the two polypeptide chains may comprise at least three parts. In some cases, the first part may be combined with the peptide conjugate / MHC complex, the second part may comprise a binding partner of a T cell surface antigen (e.g., a T cell engaging antibody), and the third part may comprise a cytokine or a fragment thereof. In some cases, the first polypeptide chain may comprise an antigen-binding domain of a peptide conjugate / MHC complex, and the second polypeptide chain may comprise an antigen-binding domain of a T cell surface antigen. In some cases, the first polypeptide chain may comprise an antigen-binding domain of a T cell surface antigen, and the second polypeptide chain may comprise an antigen-binding domain of a peptide conjugate / MHC complex. In some cases, the first polypeptide chain may comprise an antigen-binding domain of a peptide conjugate / MHC complex and a cytokine or a fragment thereof, and the second polypeptide chain may comprise an antigen-binding domain of a T cell surface antigen. In some cases, the first polypeptide chain may comprise an antigen-binding domain of a T cell surface antigen and a cytokine or a fragment thereof, and the second polypeptide chain may comprise an antigen-binding domain of a peptide conjugate / MHC complex. In some cases, the first polypeptide chain may comprise an antigen binding domain of a peptide conjugate / MHC complex, and the second polypeptide chain may comprise an antigen binding domain of a T cell surface antigen and a cytokine or a fragment thereof. In some cases, the first polypeptide chain may comprise an antigen binding domain of a T cell surface marker, and the second polypeptide chain may comprise an antigen binding domain of a peptide conjugate / MHC complex and a cytokine or a fragment thereof. In various cases, the antigen binding domain of the peptide conjugate / MHC complex and the antigen binding domain of the T cell surface antigen may be located on the same polypeptide chain, while the cytokine or a fragment thereof is fused to a different polypeptide chain.In some cases, the antigen binding domain of the peptide conjugate / MHC complex and the cytokine or fragment thereof may be located on the same polypeptide chain, and the antigen binding domain of the T cell surface antigen is located on a different polypeptide chain. In some cases, the antigen binding domain of the T cell surface antigen and the cytokine or fragment thereof may be located on the same polypeptide chain, and the antigen binding domain of the peptide conjugate / MHC complex is located on a different polypeptide chain.

[0413] The polypeptide can be a complete antibody, a bispecific antibody, a multispecific antibody, an antigen-binding (Fab) fragment, a Fab' fragment, a (Fab')2 fragment, a Fd, a Fv, a dAb, a single domain fragment or a single monomer variable antibody domain, a dual affinity retargeting (DART) molecule, a diabody (Db), a single-chain diabody (scDb), a single-chain variable fragment (scFv), a bispecific T cell engaging antibody (BiTE), a bispecific killer cell engaging antibody (BiKE), a CrossMab, a camelid antibody, a trispecific binding partner, a chimeric antigen receptor (CAR), a monomer (also known as Adnectin), a DARPin, an anticarrier protein, an affinity body, a nanobody or an antibody-like protein. In some embodiments, nanobodies are derived from the heavy chain variable domains of antibodies found in camelids (e.g., camels, llamas, alpacas). In some cases, the polypeptide can be a bispecific antibody. The bispecific antibody can be a bispecific T cell engaging antibody (BiTE). The polypeptide can include 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 CD3ε, CD3γ, CD3δ, TCRα, TCRβ, TCRγ, TCRδ, ICOS, CD28, CD27, HVEM, LIGHT, CD40, 4-1BB, OX40, DR3, GITR, CD30, TIM1, SLAM, CD2, CD4, CD8 or CD226. In some cases, the antigen binding domain of the polypeptide and the second antigen binding domain can be connected by a linker. As described above, the length of the linker can include 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. The linker can include (G4S) n or (S4G) n , where n is any integer between 1 and 10.

[0414] In various cases, the linker connecting the antigen binding domain and the second antigen binding domain of the polypeptide can affect the angle between the axis of the MHC and the axis of the polypeptide.

[0415] The binding geometry of TCR to peptide-MHC complex can be used as a descriptor of binding affinity. The diagonal binding of TCR to MHC complex can be quantified using the cross angle, such as the angle between the vector between the MHC pocket and the TCR domain. This angle corresponds to the distortion of the receptor on the peptide-MHC complex. The angle of incidence refers to the angle between the normal vector of the MHC peptide binding groove and the rotation axis between the TCR domains. This angle corresponds to the tilt of the TCR on the peptide-MHC complex and is usually measured along the XY axis. The binding geometry between the MHC complex and the polypeptide disclosed herein can be similarly measured, and methods for quantifying these binding geometries are known to those skilled in the art (Rudolph et al. How TCRs bind MHCs, peptides, and coreceptors. Annu Rev Immunol. 2006; 24: 419-66; Singh et al. Geometrical characterization of T cell receptor binding modes reveals class-specific binding to maximize access to antigen. Proteins. 2020 Mar; 88 (3): 503-513).

[0416] The binding geometry between the peptide conjugate / MHC complex and the binding partner can be characterized using the angle of incidence measured along the XY axis and / or the back angle measured along the YZ axis. In some embodiments, the binding geometry between the polypeptide and the peptide conjugate / MHC complex can be given in terms of the angle of incidence and / or the back angle. In some embodiments, the polypeptide can bind to the peptide conjugate / MHC complex at a back angle between the axis of the peptide conjugate / MHC complex and the axis of the polypeptide of about 10° to 70° (e.g., about 10°, about 20°, about 30°, about 40°, about 50°, about 60°, or about 70°). In some embodiments, the polypeptide can bind to the peptide conjugate / MHC complex at a back angle between the axis of the peptide conjugate / MHC complex and the axis of the polypeptide of about 60° to 65°. In some embodiments, the polypeptide can bind to the peptide conjugate / MHC complex at a back angle between the axis of the peptide conjugate / MHC complex and the axis of the polypeptide of about 64.7°, 67.2°, or 65.1°.

[0417] In some embodiments, the polypeptide can bind to the peptide conjugate / MHC complex at an angle of incidence between the axis of the peptide conjugate / MHC complex and the axis of the polypeptide of about 10° to 100° (e.g., about 10°, about 20°, about 30°, about 40°, about 50°, about 60°, about 70°, about 80°, about 90°, or about 100°). In some embodiments, the polypeptide can bind to the peptide conjugate / MHC complex at an angle of incidence between the axis of the peptide conjugate / MHC complex and the axis of the polypeptide of 70° to 80°. In some embodiments, the polypeptide can bind to the peptide conjugate / MHC complex at an angle of incidence between the axis of the peptide conjugate / MHC complex and the axis of the polypeptide of about 77.2°, 77.3°, or 78.4°.

[0418] In various cases described herein, the peptide conjugate / MHC complex can be presented on the cell surface. The cell can express a low copy number of the peptide conjugate / MHC complex, and wherein the low copy number is up to 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 single cell. The peptide of the peptide conjugate / MHC complex can be derived from an intracellular protein. For example, the peptide can be processed from an endogenous protein or an intracellular protein and presented on the MHC complex.

[0419] The binding partner described herein can be a T cell receptor (TCR) or a functional fragment thereof (e.g., an antigen binding fragment, a variable region, or an extracellular domain of a TCR). The binding partner described herein can be a TCR mimetic binder. The T cell receptor or a functional fragment thereof can specifically bind to the peptide conjugate / MHC complex described herein. In some cases, the TCR mimetic binder has a K of 1 or 2 for the peptide conjugate / MHC complex. D The K of a TCR for a peptide conjugate / MHC complex may be less than about 1 μM, 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 K of a TCR for a peptide conjugate / MHC complex may be less than about 1 μM, 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. DCan be less than about 1 μ M, less than about 500nM, less than about 400nM, less than about 300nM, less than about 200nM, less than about 100nM, less than about 50nM, less than about 40nM, less than about 30nM, less than about 20nM, less than about 10nM, less than about 5nM, less than about 1nM, or less than about 0.1nM, less than about 50pM, less than about 20pM, less than about 10pM, less than about 1pM, less than about 0.1pM or less. In one aspect, the present disclosure provides a method for identifying a T cell receptor (TCR) that recognizes a peptide conjugate / MHC complex described herein. In some embodiments, the method for identifying a TCR includes contacting a plurality of candidate TCRs with a peptide conjugate / MHC complex and identifying at least one TCR that binds to the peptide conjugate / MHC complex. In some embodiments, the method for identifying a TCR that recognizes a peptide conjugate / MHC complex described herein includes selecting or separating at least one TCR. Isolation of the TCR can be achieved by analyzing the binding affinity to the peptide conjugate / MHC complex. In some embodiments, the peptide-drug conjugate can be a vaccine that enhances TCR isolation. In some embodiments, the vaccine can be used to enhance G12C inhibition therapy. In some embodiments, the vaccine can be an RNA vaccine. For example, the vaccine can be an RNA vaccine that encodes a KRAS peptide comprising a G12C mutation. In some embodiments, the vaccine can be an RNA vaccine that encodes a KRAS peptide comprising a G12C mutation, which is co-administered with a KRAS peptide comprising a G12C mutation. In some embodiments, the vaccine can enhance the efficacy of, for example, G12C inhibition therapy or HapImmune TM Responsiveness of patients treated with antibodies.

[0420] In some cases, a variety of candidate TCRs are a variety of soluble TCRs. In some cases, a variety of candidate TCRs are a variety of TCRs expressed on the cell surface of a plurality of cells. In some embodiments, the method for identifying the TCR of a peptide conjugate / MHC complex as described herein includes separating or selecting cells comprising at least one TCR based on the activation markers of the cell. In some embodiments, the activation marker is a T cell marker. In some embodiments, the T cell activation marker is CD26, CD27, CD28, CD30, CD154, CD40L, CD134, CD25, CD44, CD69, CD137 or KLRG1. In another aspect, the present disclosure provides a T cell receptor (TCR) comprising at least one identified TCR for identifying a peptide conjugate / MHC complex as described herein. In some embodiments, TCR is a soluble TCR. In some embodiments, TCR is a bispecific TCR. In some embodiments, TCR is expressed on CD4+T cells. In some embodiments, TCR is expressed on CD8+T cells.

[0421] In some embodiments, the binding partner is multivalent. In some embodiments, a trispecific binding partner is provided. In some embodiments, cells express at least one section of one or more binding partners in the form of CAR. In some embodiments, the binding partner of the present disclosure can be provided as a complex with a polynucleotide (e.g., RNA polynucleotide) to form an aptamer. In some embodiments, the multivalent binding partner includes a binding component (e.g., a paratope) that imparts specificity to a specific target (e.g., any cancer cell marker) on a desired cell type. In some embodiments, a trispecific leukocyte engaging antibody is provided. In some embodiments, the binding partner can be part of a molecule that is activated only in the presence of a protease or other enzyme present in the tumor microenvironment, such embodiments relating to, for example, 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, wherein the descriptions of probodies and protease activation are incorporated herein by reference. In some embodiments, the present disclosure provides universal haptens that can be grafted onto inhibitors.

[0422] In some embodiments, the CAR of the present disclosure includes scFv, which includes heavy and light chain variable regions as described herein. As known in the art for previously described CARs, scFv is present in a continuous polypeptide that also includes a CD3 ζ chain and a costimulatory domain. In some embodiments, the continuous polypeptide also includes a CD3 γ chain or a CD3 ε chain. In some embodiments, the costimulatory domain includes a 4-1BB costimulatory domain or a CD28 costimulatory domain. CAR may also include a co-receptor hinge sequence, such as a CD8 co-receptor hinge sequence.

[0423] In some embodiments, the binding partners of the present disclosure may comprise a constant region, such as an Fc region. Any isotype of the constant region may be included. Binding partners comprising a constant region may be particularly suitable for antibody-dependent cell-mediated cytotoxicity (ADCC) or antibody-dependent cellular phagocytosis (ADCP), and thus may kill target cells by a cell-mediated reaction of any of a variety of effector cells. Similarly, the constant region may be particularly suitable for enhancing complement-mediated reactions.

[0424] In some embodiments, the binding partner of the present disclosure can be modified so that it is present in a fusion protein. In some embodiments, the antigen-binding fragment of the binding partner can be present in the fusion protein, and / or the constant region can be a component of the fusion protein. In some embodiments, the fusion protein comprises amino acids from at least two different proteins. The fusion protein can be produced using any of a variety of standard molecular biology methods, including but not necessarily limited to expressing from any suitable expression vector. In some embodiments, the binding partner described herein can be present in the fusion protein together with a detectable protein (e.g., green fluorescent protein (GFP), enhanced GFP (eGFP), mCherry, etc.). In some embodiments, as an alternative to the expression vector, the mRNA or chemically modified mRNA encoding any binding partner described herein can be delivered to the cell so that the binding partner is translated by the cell. In some embodiments, the fusion protein comprises a binding partner disclosed herein and a cytokine. In some embodiments, the cytokine is IL-2, IL-7, IL-15, IL-12, IL-18 or IL-21. In some embodiments, the cytokine is modified to increase at least one therapeutic characteristic, including but not limited to bioavailability, efficacy, half-life extension or other desired characteristics. 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.

[0425] In some embodiments, the binding partner of the present disclosure can be fused with another antibody. For example, the binding partner of the present disclosure can be fused with an antibody or fragment thereof as an immune checkpoint inhibitor. In some embodiments, the antibody or fragment thereof as an immune checkpoint inhibitor can be an 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-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 or an agonist anti-CD28 antibody.

[0426] In some embodiments, the binding partners described herein are used to carry drugs or toxins, and thus the binding partners can be provided as immunotoxins or in the form of antibody-drug conjugates (ADCs).

[0427] In some embodiments, the agent used to generate the immunotoxin comprises an enzymatically active toxin and an enzymatically active fragment thereof. Suitable enzymatically active toxins include, but are not limited to, diphtheria toxin A chain, non-binding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, α-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, jatropha toxin, crotonin, saponin inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, and tricothecenes. These toxins can be provided as components of fusion proteins or can be covalently linked to a binding partner by any suitable conjugation method.

[0428] The binding partner can be connected to a chemotherapeutic agent using any suitable linker to form an antibody drug conjugate (ADC). In some embodiments, the linker includes a disulfide, a hydrazine, or a thioether. In some embodiments, the ADC comprises two payloads (e.g., two different chemotherapeutic agents), each of which is connected to the binding partner via a linker. In some embodiments, the ADC comprises two payloads (e.g., two different chemotherapeutic agents), each of which is connected to the binding partner via a linker using a site-specific aldehyde tag. The chemotherapeutic agent can be reversibly or irreversibly attached to the binding partner.

[0429] Cleavable linkers may be particularly useful for killing bystander cells. In some embodiments, a protease recognition site may be included to release the chemotherapeutic agent from the binding partner by action of a protease that recognizes and cleaves the protease recognition site. Thus, the ADC may be considered to comprise a prodrug.

[0430] In some embodiments, the binding partner of the present disclosure may include a linker sequence. As a non-limiting example, ScFv may include a linker that connects the fragments comprising the paratope to each other. Suitable amino acid linkers may be mainly composed of relatively small neutral amino acids (e.g., glycine, serine, and alanine), and may include multiple copies of sequences rich in glycine and serine. In specific non-limiting embodiments, the linker includes 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 amino acids. In one example, the linker may be a glycine-serine-alanine linker G4SA3 or a glycine-serine linker (G4S)4 linker. In some embodiments, the binding partner may include a cell localization signal or a secretion signal. In some embodiments, the binding partner may include a transmembrane domain, and therefore may be transported to the cell membrane and anchored in the cell membrane. For secretion, any suitable secretion signal may be used, and many secretion signals are known in the art.

[0431] In some embodiments, the binding partner can be part of the ADC, and thus the binding partner comprises a drug. The drug may include but is not necessarily limited to any suitable chemotherapeutic agent. In some embodiments, the ADC comprises a binding partner and a chemotherapeutic agent, and the chemotherapeutic agent is an anti-microtubule agent, an alkylating agent, or a DNA minor groove binder. In some embodiments, the chemotherapeutic agent includes a maytansinoid, a dolastatin, an auristatin analog, or cryptophycin. In some embodiments, the chemotherapeutic agent is a duocarmycin derivative or an antibiotic, such as an enediyne antibiotic or a pyrrolobenzodiazepine (PBD, pyrolobenodiazepine), including its dimer. In some embodiments, the chemotherapeutic agent is an enzyme inhibitor, such as a topoisomerase or polymerase inhibitor. In some embodiments, the chemotherapeutic agent includes doxorubicin or a metal-containing compound, such as a platinum-containing compound, non-limiting examples of which include cisplatin, carboplatin, or oxaliplatin. In some embodiments, the ADC comprises a binding partner described herein and any of the drugs described in Barf and Kaptein, dx.doi.org / 10.1021 / jm3003203, J. Med. Chem. 2012, 55, 6243-6262, or Wilson et al., dx.doi.org / 10.1021 / jm400224q, J. Med. Chem. 2013, 56, 7463-7476, Lambert and Morris, Adv Ther (2017) 34: 1015-1035, or Tarantino and Tolaney, Cancer Res (2022) 82(20): 3659-3661, wherein the descriptions of drugs used as components of the ADC are incorporated herein by reference. In some embodiments, the binding partner is conjugated to a cytokine or otherwise includes a cytokine, including but not 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), thereby providing a cytokine conjugate. In some embodiments, the binding partner comprises a toxin conjugate. The toxin conjugate may include but not be limited to Pseudomonas exotoxin A (PE), diphtheria toxin (DT) or a recombinant variant thereof. In some embodiments, the binding partner may include an immunotoxin agent derived from PE or DT, including but not limited to BL22, LMB-2, CAT-8015, SS1P, MR1-1 or Zemab.

[0432] In order to produce a binding partner, any suitable expression system can be used. Generally, the polynucleotide encoding the binding partner is used to express the binding partner in any suitable cell system, and its non-limiting embodiments include NS0 mouse myeloma cells, human cell lines and Chinese hamster ovary (CHO) cells. In some embodiments, the disclosure provides polynucleotides that can selectively hybridize with the polynucleotides encoding any CDR or CDR combinations as described herein. In some embodiments, the polynucleotides selectively hybridize with the heavy chain CDR1, CDR2 and CDR3 of any of the binding partners described in the coding. In some embodiments, the polynucleotides selectively hybridize with the light chain CDR1, CDR2 and CDR3 of any of the binding partners described in the coding. In some embodiments, the polynucleotides selectively hybridize with the polynucleotides encoding the CDR1, CDR2 and CDR3 of the heavy chain and light chain of any of the binding partners described in the coding.

[0433] In some embodiments, the binding partner described herein can be a component of a fusion protein. In some embodiments, for example, for a binding partner produced as a fusion protein, a peptide linker can be used. In some embodiments, the peptide linker comprises any self-cleavage signal. In some embodiments, the self-cleavage signal can be present in an open reading frame (ORF) identical to the ORF encoding the binding partner. The self-cleavage amino acid sequence is generally about 18-22 amino acids long. Any suitable sequence can be used, non-limiting examples of which include: T2A (EGRGSLLTCGDVEENPGP); P2A (ATNFSLKQAGDVENPGP); E2A (QCTNYALKLAGDVESNPGP) and F2A (VKQTLNFDLKLAGDVESNPGP).

[0434] If any fragment of a protein comprising a binding partner described herein is a component of a library, including but not limited to a phage display library or a yeast surface display library, the present disclosure includes the following proviso: the binding partner may not contain any fragment of the library that contains a phage or yeast amino acid sequence (including but not limited to a phage coat protein or a yeast host protein, including but not limited to Aga2). Thus, in certain embodiments, a binding partner may be present in a fusion protein, but the fusion protein does not contain a phage coat protein. In some embodiments, any binding partner described herein may not contain any pill phage coat protein, or any portion of a protein from M1, fd filamentous phage, T4, T7, or lambda phage.

[0435] In some embodiments, the binding partners of the present disclosure comprise a detectable label that can be used for diagnostic or therapeutic purposes. For example, a detectable label can be used to locate the binding partner for pathological and / or in vivo imaging methods. In some embodiments, the binding partner is conjugated to any of a variety of radioactive agents, including but not limited to highly radioactive atoms such as In111, At211, I131, I125, Y90, Re186, Re188, Sm153, Bi212, P32, Pb212, and radioisotopes of Lu. In some specific embodiments, for example for imaging, the binding partner can be conjugated to a radioactive atom for scintigraphy methods, such as 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, O17, or gadolinium (III) or manganese (II). In some embodiments, the radioactive agent is suitable for CAT scanning or PET imaging. In some embodiments, indium 111, technetium 99 or iodine 131 can be used for planar scanning or single photon emission computed tomography (SPECT). Positron emission markers such as fluorine 19, iodine 123 and iodine 124 can be used for positron emission tomography. Paramagnetic ions such as gadolinium (III) or manganese (II) can be used for magnetic resonance imaging MRI. In some embodiments, the radioactive isotope attached to the binding partner can also be used for therapeutic methods. In some embodiments, the radioactive agent or isotope includes alpha-emitting radionuclides. In some embodiments, the radioactive agent or isotope includes beta-emitting radionuclides. In some embodiments, the present disclosure provides antibodies of the present technology conjugated to diagnostic agents or therapeutic agents. Diagnostic agents can include non-radioactive labels, contrast agents (e.g., for magnetic resonance imaging, computed tomography or ultrasound) and / or radioactive labels, and the radioactive labels can be gamma, beta, alpha, Auger electrons or positron emitting isotopes. Diagnostic agents are molecules that are administered conjugated to an antibody portion (ie, an antibody or antibody fragment or subfragment) and can be used to diagnose or detect disease by localizing cells containing the antigen.

[0436] The binding partner can be a polypeptide. The polypeptide can bind to the peptide conjugate / MHC complex. The three-dimensional structure of the polypeptide bound to the peptide conjugate / MHC complex can be determined by various methods, such as crystallography, cryoEM, or NMR. The polypeptide can be in contact with the peptide conjugate / MHC complex with an interface area of ​​at least about 500, 600, 700, 800, 900, 1,000, 1,200, 1,500, 2,000, or more. The surface area of ​​residues buried during the interaction between a polypeptide and a peptide conjugate / MHC complex can be calculated using PDBePISA. A polypeptide may contact the MHC of a peptide conjugate / MHC complex with an interfacial area greater than the interfacial area with the peptide or a targeted covalent inhibitor. A polypeptide may form a binding pocket at the interface between the VH and VL domains to accommodate a targeted covalent inhibitor of the peptide conjugate / MHC complex. A polypeptide may not bind to a peptide conjugate / MHC complex via head-to-head coaxial interactions. In some cases, a polypeptide may bind to a peptide conjugate / MHC complex via head-to-head coaxial interactions. For example, a polypeptide may bind to a conjugate / MHC complex in a manner similar to how a TCR binds to a peptide / MHC complex. A polypeptide may bind to a peptide conjugate / MHC complex at an angle of approximately 10° to 60°, approximately 10° to 70°, or approximately 10° to 80° between the axis of the MHC and the axis of the polypeptide. A polypeptide may bind to a peptide conjugate / MHC complex at an angle of approximately 40° between the axis of the MHC and the axis of the polypeptide.

[0437] The polypeptide may contact the α1 domain and the α2 domain of the MHC heavy chain. The polypeptide may bind to an epitope of the MHC. The epitope may comprise one or more residues from a region comprising residues 62-66, 106-109, and / or 150-170 of the MHC. The polypeptide may bind to an epitope of the MHC that may comprise one or more residues from a region comprising residues 62-66, 106-109, and / or 150-170 of HLA-A*03:01 or HLA-A*11:01. The polypeptide may bind to an epitope of the MHC that may 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 HLA-A*03:01. The polypeptide may bind to an epitope of MHC that may 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 HLA-A*03:01. The polypeptide may bind to an epitope of MHC that may comprise one or more residues selected from the group consisting of residues 62, 65, 66, 150, 151, 154, 155, 157, and 158 of HLA-A*03:01. The polypeptide may bind to an epitope of MHC that may comprise one or more residues selected from the group consisting of residues 62, 65, 66, 150, 151, 154, 155, 157, and 158 of HLA-A*03:01. The polypeptide may bind to an epitope of MHC that may 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 HLA-A*11:01. The polypeptide may bind to an epitope of MHC that may 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 HLA-A*11:01. The polypeptide may bind to an epitope of MHC that may comprise one or more residues selected from the group consisting of residues 62, 65, 66, 151, 154, 155, and 158 of HLA-A*11:01. The polypeptide may bind to an epitope of MHC, and the epitope may comprise one or more residues selected from the group consisting of residues 62, 65, 66, 151, 154, 155, and 158 of HLA-A*11:01.

[0438] In some aspects, the present disclosure provides methods for identifying or designing polypeptides that can bind to a peptide conjugate / MHC complex based on the three-dimensional structure of the peptide conjugate / MHC complex. The three-dimensional structure of the peptide conjugate / MHC complex can be obtained from an existing complex structure of a binding partner that binds to the peptide conjugate / MHC complex (e.g., a structure provided herein). Computer-assisted methods can be used to identify or design polypeptides that can be linked to a targeted covalent inhibitor or a fragment thereof, wherein the targeted covalent inhibitor or a fragment thereof is covalently linked to a peptide of the peptide conjugate / MHC complex. In some embodiments, the method for identifying or designing a polypeptide candidate that can be linked to a targeted covalent inhibitor comprises: (a) providing Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and Figure 61 The method may further comprise (i) providing the coordinates of at least two atoms of the peptide conjugate / MHC complex. The method may further comprise (b) providing the structure of a candidate polypeptide comprising an antigen binding domain for binding to the peptide conjugate / MHC complex. The method may further comprise (c) fitting the structure of the candidate polypeptide to at least two atoms of the peptide conjugate / MHC complex. In some cases, fitting may comprise determining interactions between one or more atoms of the antigen binding domain of the candidate polypeptide and atoms of the peptide conjugate / MHC complex. The method may further comprise (d) selecting the candidate polypeptide if the candidate polypeptide is predicted to bind to the peptide conjugate / MHC complex.

[0439] Steps (a) and (b) of the computer-assisted method for designing a polypeptide that can be linked to a targeted covalent inhibitor can occur in any order or simultaneously. For example, in some embodiments, the computer-assisted method for designing a polypeptide that can be linked to a targeted covalent inhibitor comprises the following steps: (a) providing a structure of a candidate polypeptide comprising an antigen binding domain for binding to a peptide conjugate / MHC complex; (b) providing Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and Figure 61 (c) fitting the structure of the candidate polypeptide to the at least two atoms of the peptide conjugate / MHC complex and the at least two atoms of the antigen binding domain of the polypeptide bound to the peptide conjugate / MHC complex, wherein the fitting comprises determining interactions between one or more atoms of the antigen binding domain of the candidate polypeptide and atoms of the peptide conjugate / MHC complex; and (d) selecting the candidate polypeptide if the candidate polypeptide is predicted to bind to the peptide conjugate / MHC complex.

[0440] In some embodiments of a computer-assisted method for designing a polypeptide that can be linked to a targeted covalent inhibitor, interactions between atoms of an antigen binding domain and atoms of a peptide are determined. For example, such a method may include: (a) providing a structure of a polypeptide comprising an antigen binding domain that binds 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 the antigen binding domain, wherein the one or more amino acid substitutions are residues of the antigen binding domain of the polypeptide that comprise one or more atoms that interact with or modulate the interaction with one or more atoms of the peptide conjugate / MHC complex; and (d) selecting the candidate polypeptide if it binds to the peptide conjugate / MHC complex or is predicted to bind to the peptide conjugate / MHC complex with a higher affinity than the affinity of the polypeptide comprising the antigen binding domain to the peptide conjugate / MHC complex.

[0441] In some cases, a computer-assisted method for identifying or designing a polypeptide that can be linked to a targeted covalent inhibitor comprises using a computer system. The computer system can be, for example, a programmed computer comprising a processor, a data storage system, an input device, and an output device. In some embodiments, the steps of such a method include: (a) inputting a polypeptide comprising a polypeptide from or belonging to Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and Figure 61 (b) using the processor to compare the data set with a computer database of structures of polypeptides containing antigen binding domains stored in the computer data storage system, the polypeptides binding or presumed to bind or expected to bind to the peptide conjugate / MHC complex; (c) using computer methods to select structures from the database that may bind to certain peptide conjugate / MHC complexes; (d) using computer methods to construct models of the selected structures; and (e) outputting the selected structures to the output device. Optionally, one or more selected structures can be synthesized, and the synthesized structures can be tested to see if they are peptide conjugate / MHC complexes.

[0442] Computer readable media can be used to identify or design polypeptide candidates comprising an antigen binding domain that binds to a peptide conjugate / MHC complex. For example, in some embodiments, such a computer readable medium can comprise: (a) a computer program according to Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and Figure 61 wherein the data defines or provides a three-dimensional structure of a polypeptide comprising an antigen binding domain that is bound to a peptide conjugate / MHC complex, and wherein the peptide conjugate of the peptide conjugate / MHC complex is a peptide covalently linked to a targeted covalent inhibitor, or a fragment thereof, or at least one subdomain thereof; or (b) structure factor data for a polypeptide comprising an antigen binding domain that is bound to a peptide conjugate / MHC complex, wherein the structure factor data can be obtained from Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and Figure 61 In other embodiments, the computer readable medium may include: (a) according to Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and Figure 61 wherein the data defines the three-dimensional structure of or provides the structure of 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 or at least one subdomain thereof; or (b) structure factor data for the peptide conjugate / MHC complex, wherein the structure factor data can be obtained from Figures 57A-57C 、 Figure 58A-Figure 58B 、 Figures 59A-59C 、 Figures 60A-60B and Figure 61 The atomic coordinate data of any one of the structures is obtained.

[0443] Any binding partner described herein can be fully or partially humanized. The techniques for humanizing antibodies are known in the art and can be applied to the present disclosure. In some embodiments, humanization can be performed, for example, by CDR transplantation. In some embodiments, in order to humanize or otherwise improve the properties of the binding partner, one or more amino acids in the variable region can be changed. In some embodiments, one or more amino acids in the framework region can be changed.

[0444] The present disclosure includes binding partners for diagnosis and treatment methods. For treatment methods, in certain embodiments, the binding partner can be delivered as mRNA or DNA polynucleotides encoding the binding partner. It is believed that administering DNA or RNA encoding any binding partner described herein is also a method for delivering such a binding partner to an individual or one or more cells. In some embodiments, mRNA includes synthetic mRNA containing modified nucleotides. In some embodiments, modification can include N1-methyl-pseudouridine (1mΨ) nucleotide replacement and / or 5-methylcytidine (m5C) replacement. Methods for delivering protein-encoding DNA and RNA are known in the art and can be suitable for delivering binding partners, giving the benefits of the present disclosure. In some embodiments, one or more expression vectors are used and include viral vectors. Therefore, in some embodiments, viral expression vectors are used. Viral expression vectors can be used as naked polynucleotides, or can include any viral particles, including but not limited to defective interfering particles or other replication-defective viral constructs and virus-like particles. In some embodiments, the expression vector includes modified viral polynucleotides, for example, from adenovirus, herpes virus or retrovirus. In some embodiments, retroviral vectors adapted from murine Moloney leukemia virus (MLV), or lentiviral vectors, such as those adapted from human immunodeficiency virus type 1 (HIV-1), may be used.

[0445] In one embodiment, an oncolytic viral vector is used. Oncolytic viruses (oV) (including cowpox (OVV)) mediate anti-cancer effects by directly lysing the tumor and stimulating the innate immune response by producing damage-associated molecular patterns (DAMPs) and the presence of viral-derived pathogen-associated molecular patterns (PAMPs), resulting in increased production of type I interferon. In addition, OVV-mediated oncolysis can promote host antigen-presenting cells in the tumor microenvironment to directly acquire tumor-derived antigens, resulting in improved T cell pre-sensitization and coordination of the anti-tumor immune response effector phase. In alternative embodiments, a recombinant adeno-associated virus (AAV) vector can be used. In certain embodiments, the expression vector is a self-complementary adeno-associated virus (scAAV).

[0446] Pharmaceutical preparations containing binding partners are included in the present disclosure and can be prepared by mixing them with one or more pharmaceutically acceptable carriers. Pharmaceutically acceptable carriers include solvents, dispersion media, isotonic agents, etc. The carrier can be a liquid, a semisolid, such as a paste, or a solid carrier. Examples of carriers include water, saline solutions or other buffers (e.g., phosphate, citrate buffer), oils, alcohols, proteins (e.g., serum albumin, gelatin), carbohydrates (e.g., monosaccharides, disaccharides and other carbohydrates, including glucose, sucrose, trehalose, mannose, mannitol, sorbitol or dextrin), gels, lipids, liposomes, resins, porous matrices, adhesives, fillers, coatings, stabilizers, preservatives, liposomes, antioxidants, chelating agents (e.g., EDTA), salt-forming counterions (e.g., sodium); nonionic surfactants (e.g., TWEEN, PLURONICS or polyethylene glycol (PEG)) or combinations thereof. In some embodiments, liposome preparations comprising one or more binding partners are provided. Liposomal preparations include, but are not limited to, liposome nanoparticles.

[0447] In some embodiments, an effective amount of one or more binding partners is administered to an individual in need thereof. In some 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 can also inhibit or prevent the onset of a disease or the recurrence of a disease. An individual physician can select the exact dosage based on the patient to be treated. Dosage and administration can be adjusted to provide sufficient levels of the binding partner to maintain the desired effect. Other factors that may be considered include the severity and type of the disease state, the patient's age, weight, and sex, the desired duration of treatment, method of administration, time and frequency of administration, drug combination, reaction sensitivity, and / or tolerance / response to treatment.

[0448] Binding partners and pharmaceutical compositions comprising binding partners can be administered to an individual in need thereof using any suitable route, examples of which include intravenous, intramuscular, intraperitoneal, intracerebrospinal, subcutaneous, intraarticular, intrasynovial, oral, topical, or inhalation routes, depending on the specific condition being treated. The composition can be administered parenterally or enterally. The composition can be introduced as a single administration or multiple administrations, or can be introduced in a continuous manner over a period of time. For example, administration can be a pre-specified number of administrations or daily, weekly, or monthly administrations, which can be continuous or intermittent, as indicated by the treatment.

[0449] In some embodiments, the individual in need of the compositions of the present disclosure has been diagnosed with or is suspected of having cancer. In some embodiments, the cancer is a solid tumor or a hematological malignancy. In some 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 (or colorectal cancer), esophageal cancer, glioma, glioblastoma or other brain cancer, gastric cancer, bladder cancer, testicular cancer, head and neck cancer, thyroid cancer, adrenal cancer, melanoma or other 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 needs to treat a neuroendocrine tumor. In some embodiments, the individual needs to treat any pre-tumor disease, including myelodysplastic syndrome or myeloproliferative tumor. In some embodiments, the binding partner is used to prevent any of the types of cancer. In some embodiments, the individual in need has been treated with a previous treatment, and the individual is refractory to the previous treatment. In some embodiments, the cancer is a relapsed or refractory cancer.

[0450] In some embodiments, one or more binding partners (including but not necessarily limited to pharmaceutical preparations) are administered to an individual in need thereof to demonstrate improved activity relative to a control. In some embodiments, the control comprises different antibodies, different forms of the same antibody / binding partner, or an antibody / binding partner delivered without the addition of additional agents. In some embodiments, the binding partners described herein provide improved antibody-dependent...

Claims

1. 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 a covalent reaction of a targeted covalent inhibitor or a fragment thereof with a peptide; and (b) MHC.

2. A binding partner according to claim 1, wherein the binding affinity of the binding partner to the peptide conjugate / MHC complex is greater than the binding affinity to the peptide or free targeted covalent inhibitor.

3. A binding partner according to claim 1 or 2, wherein 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.

4. The binding partner of any one of claims 1-3, wherein the MHC is a human leukocyte antigen (HLA), optionally wherein the HLA is HLA-A, HLA-B or HLA-C.

5. The binding partner of claim 4, wherein the HLA molecule is an HLA-A*02:01, HLA-A*03:01, HLA-A*01:01, HLA-A*11:01, HLA-A*24:02, HLA-A*26:01, HLA-B*07:02, HLA-B*08:01, HLA-B*27:05, HLA-B*39:01, HLA-B*40:01, HLA-B*58:01 and / or HLA-B*15:01 molecule.

6. The binding partner of any one of claims 1 to 5, wherein the peptide comprises a nucleophilic or electrophilic residue, which residue is optionally one of cysteine, lysine, tyrosine, histidine, serine, arginine or threonine.

7. The binding partner of any one of claims 1 to 6, wherein the peptide comprises a cysteine ​​residue.

8. The binding partner of any one of claims 1-7, wherein the peptide conjugate is formed by a covalent reaction between the targeted covalent inhibitor and a cysteine ​​residue in the peptide.

9. The binding partner of any one of claims 1-8, wherein the peptide is a fragment of a protein associated with cancer, optionally wherein the protein is encoded by a gene that is mutated in cancer.

10. The binding partner of any one of claims 1-9, wherein the peptide is a fragment of an enzyme, and wherein the targeted covalent inhibitor is an inhibitor of the enzyme.

11. The binding partner of claim 10, wherein the enzyme is a kinase or a GTPase.

12. A binding partner according to any one of claims 1-11, wherein the peptide is or is derived from RAS, Bruton's tyrosine kinase (BTK), any epidermal growth factor receptor (EGFR) family member selected from EGFR (ERBB1), HER2 / NEU (ERBB2), HER3 (ERBB3) and HER4 (ERBB4); MET (HGFR); any fibroblast growth factor receptor (FGFR); any cyclin-dependent kinase (CDK); acetylcholinesterase (ACHE); p90 ribosomal S6 kinase (RSK); TP53, IDH1, GNAS, FBXW7, CTNNB1, DNMT3A, a cathepsin selected from cathepsin B, C, F, H, K, L, O, S, V, W and X; any caspase; a protein involved in obesity, which is optionally pancreatic lipase or METAP2; any cancer testis antigen (CTA); long interspersed element-1 (LINE-1); short interspersed elements, which are optionally Alu; and any endogenous retroviral protein, and optionally wherein the RAS is KRAS, HRAS or NRAS.

13. The binding partner of any one of claims 1-12, wherein the peptide comprises KRAS G12C 、KRAS G12D 、KRAS G12R 、KRAS G12S 、HRAS G12C 、HRAS G12D 、HRAS G12R 、HRAS G12S 、NRAS G12C NRAS G12D NRAS G12R or NRAS G12S Fragment of .

14. The binding partner of any one of claims 1-13, wherein the peptide comprises the amino acid sequence of VVVGACGVGK, VVGACGVGK or KLVVVGACGV.

15. The binding partner of any one of claims 1-14, wherein the targeted covalent inhibitor is (i) a ternary complex KRAS G12C Inhibitors or KRAS G12C Degraders, (ii) Ternary complex KRAS G12D Inhibitors or KRAS G12D Degraders, (iii) ternary complex KRAS G12R Inhibitors or KRAS G12R Degraders, or (iv) ternary complex KRAS G12S Inhibitors or KRAS G12S Degradation agent.

16. The binding partner of claim 13, wherein (i) the peptide comprises KRAS G12C mutation, and the targeted covalent inhibitor is KRAS G12C inhibitor, (ii) the peptide comprises KRAS G12D mutation, and the targeted covalent inhibitor is KRAS G12D inhibitor, (iii) the peptide comprises KRAS G12R mutation, and the targeted covalent inhibitor is KRAS G12R inhibitor, or (iv) the peptide comprises KRAS G12S mutation, and the targeted covalent inhibitor is KRAS G12S Inhibitor.

17. The binding partner of any one of claims 13-16, wherein the targeted covalent inhibitor is osimertinib, ibrutinib, neratinib, sotolacib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1, or BI 182391.

18. The binding partner of any one of claims 1-12, wherein the peptide comprises a fragment of EGFR.

19. The binding partner of any one of claims 1-12 or 17, wherein the peptide comprises the amino acid sequence of QLMPFGCLL, LMPFGCLLDY or MPFGCLLDY.

20. The binding partner of any one of claims 1-12, wherein the peptide comprises a fragment of EGFR and the targeted covalent inhibitor is an inhibitor of an EGFR family kinase.

21. The binding partner of any one of claims 18-20, wherein the targeted covalent inhibitor is osimertinib or neratinib.

22. The binding partner of any one of claims 1-12, wherein the peptide comprises a fragment of BTK.

23. The binding partner of any one of claims 1-12 or 22, wherein the peptide comprises the amino acid sequence of YMANGCLLNY.

24. The binding partner of claim 22 or 23, wherein the targeted covalent inhibitor is ibrutinib.

25. The binding partner of any one of claims 1-24, wherein the peptide is a full-length protein that is processed in the cell after covalent reaction with the targeted covalent inhibitor to produce smaller peptide fragments.

26. The binding partner of any one of claims 1-17, wherein the peptide conjugate comprises a compound selected from the group consisting of compounds 1-8: , It is covalently bound to a cysteine ​​residue in a peptide comprising the amino acid sequence of VVVGACGVGK, VVGACGVGK or KLVVVGACGV.

27. The binding partner of any one of claims 13-17, 25 or 26, wherein the MHC is HLA-A*02:01, HLA-A*03:01 and / or HLA-A*11:

01.

28. The binding partner of claim 26 or 27, wherein the peptide comprises the amino acid sequence of VVVGACGVGK or VVGACGVGK, and the MHC is HLA-A*03:01 or HLA-A*11:

01.

29. A binding partner according to claim 26 or 27, wherein the peptide comprises the amino acid sequence of KLVVVGACGV and the MHC is HLA-A*02:

01.

30. The binding partner of any one of claims 1-12 or 18-21, wherein the peptide conjugate comprises compound 9: It is covalently bound to a cysteine ​​residue in a peptide comprising the amino acid sequence of QLMPFGCLL, LMPFGCLLDY or MPFGCLLDY.

31. The binding partner of claim 30, wherein the MHC is HLA-A*02, HLA-A*01, HLA-A*03 or HLA-A*26.

32. The binding partner of any one of claims 1-12 or 22-24, wherein the peptide conjugate comprises compound 10: It is covalently bound to a cysteine ​​residue in a peptide comprising the amino acid sequence of YMANGCLLNY.

33. The binding partner of claim 32, wherein the MHC is HLA-A*01:

01.

34. A binding partner according to any one of claims 1-17 or 25-29, wherein the binding partner specifically binds to a peptide conjugate / MHC complex, wherein the peptide conjugate is composed of sotolacib and KRAS G12C The binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: (i) CDR-H1, which comprises the amino acid sequence of DYSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) CDR-H2 comprising the amino acid sequence of SISSSSGSTSYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and / or (iii) CDR-H3 comprising an amino acid sequence of GX1WX2X3AMDY, wherein X1 is G, R, H, S or K, X2 is Y or I, and X3 is P or A: and / or (b) the VL comprises: (i) CDR-L1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) CDR-L2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and / or (iii) CDR-L3, which comprises the amino acid sequence of QQX1SYVX2X3X4IT, wherein X1 is I, A, P, V or S, X2 is K, R, A or H, X3 is K or R, and X4 is L, T, K, R, V, A or E.

35. A binding partner according to any one of claims 1-17 or 25-29, wherein the binding partner specifically binds to a peptide conjugate / MHC complex, wherein the peptide conjugate is composed of sotolacib and KRAS G12C The invention relates to a method for preparing an oligonucleotide comprising the steps of: reacting the oligonucleotides to form a mononucleotide binding protein, wherein the binding partner comprises CDR-H1, CDR-H2 and CDR-H3 amino acid sequences of the VH amino acid sequence and / or CDR-L1, CDR-L2 and CDR-L3 amino acid sequences of the VL amino acid sequence of the binding partner selected from the group consisting of: RA_D11, RA_D01-RA_D04, RA_D06-RA_D09, RA_D12-RA_D14, RA_D16, RA_D18-RA_D21, RA_D23 and RA_D24; or a variant thereof comprising 1-5 amino acid changes in one or more CDR amino acid sequences.

36. A binding partner according to claim 34 or 35, wherein the binding partner comprises a CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and / or CDR-L3 amino acid sequence of a binding partner selected from the group consisting of: RA_D11, RA_D01-RA_D04, RA_D06-RA_D09, RA_D12-RA_D14, RA_D16, RA_D18-RA_D21, RA_D23 and RA_D24 (Table G and Table H), or a variant thereof comprising 1-5 amino acid changes in one or more CDR amino acid sequences.

37. The binding partner of any one of claims 34-36, wherein the binding partner comprises a VH and / or VL amino acid sequence that is 90%, 95% or 100% identical to the following VH and VL sequences:

38. A binding partner according to any one of claims 1-12, 18-21, 30 or 31, wherein the binding partner specifically binds to a peptide conjugate / MHC complex, wherein the peptide conjugate is formed by a covalent reaction of osimertinib and an EGFR peptide, wherein the binding partner comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein (a) the VH comprises: (i) CDR-H1, which comprises the amino acid sequence of SSYIH, or a variant thereof comprising 1-5 amino acid changes; (ii) CDR-H2 comprising the amino acid sequence of YISPSYGSTSYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and / or (iii) CDR-H3 comprising an amino acid sequence of EX1X2X3MX4X5DY, wherein X1 is Y, L, S or E, X2 is V, T or I, X3 is T or I, X4 is A, T or S, and X5 is L, A, I, K, P or T; and / or (b) the VL comprises: (i) CDR-L1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) CDR-L2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes; and / or (iii) CDR-L3, which comprises the amino acid sequence of QQYX1X2WPX3T, wherein X1 is S or A, or S; X2 is Y, H, A, D, E, K, S or G; and X3 is I or E.

39. A binding partner according to any one of claims 1-12, 18-21, 30 or 31, wherein the binding partner specifically binds to a peptide conjugate / MHC complex, wherein the peptide conjugate is formed by the covalent reaction of osimertinib with an EGFR peptide, wherein the binding partner comprises CDR-H1, CDR-H2 and CDR-H3 amino acid sequences of the VH amino acid sequence and / or CDR-L1, CDR-L2 and CDR-L3 amino acid sequences of the VL amino acid sequence of the binding partner selected from the group consisting of: OEA2-5, EO_Q01-EO_Q18 and EO_Q20-EO_Q24, or variants thereof comprising 1-5 amino acid changes in one or more CDR amino acid sequences.

40. A binding partner according to claim 38 or 39, wherein the binding partner comprises the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and / or CDR-L3 amino acid sequences of a binding partner selected from the group consisting of: OEA2-5, EO_Q01-EO_Q18 and EO_Q20-EO_Q24 (Table I and Table J), ​​or a variant thereof comprising 1-5 amino acid changes in one or more CDR amino acid sequences.

41. The binding partner of any one of claims 38-40, wherein the binding partner comprises a VH and / or VL amino acid sequence that is 90%, 95% or 100% identical to the following VH and VL sequences:

42. A binding partner according to any of the preceding claims, wherein the binding partner has a higher affinity for a peptide conjugate / MHC complex comprising a first HLA than for a peptide conjugate / MHC complex comprising a second HLA.

43. A binding partner according to claim 42, wherein the first HLA and the second HLA are each selected from the group consisting of HLA-A*02:01, HLA-A*03:01, HLA-A*01:01, HLA-A*11:01, HLA-A*24:02, HLA-A*26:01, HLA-B*07:02, HLA-B*08:01, HLA-B*27:05, HLA-B*39:01, HLA-B*40:01, HLA-B*58:01 and / or HLA-B*15:

01.

44. A binding partner according to any one of claims 1-43, wherein the binding partner is a complete antibody, a bispecific antibody, a multispecific antibody, an antigen binding (Fab) fragment, a Fab' fragment, a (Fab')2 fragment, Fd, Fv, dAb, a single domain fragment or a single monomer variable antibody domain, a dual affinity retargeting (DART) molecule, a diabody (Db), a single chain diabody (scDb), a single chain variable fragment (scFv), a bispecific T cell engaging antibody (BiTE), a bispecific killer cell engaging antibody (BiKE), a CrossMab, a camelid antibody, a trispecific binding partner, a chimeric antigen receptor (CAR), a monomer (also known as Adnectin), a DARPin, an anticalin, an affibody or an antibody-mimicking protein.

45. The binding partner of any one of claims 1-44, wherein the binding partner is bispecific.

46. ​​A binding partner according to claim 45, wherein the binding partner specifically binds to the peptide conjugate / MHC complex and a T cell antigen.

47. A binding partner according to claim 45 or 46, wherein the binding partner specifically binds the peptide conjugate / MHC complex and human CD3.

48. The binding partner of claim 47, wherein the binding partner comprises the VH and / or VL amino acid sequence of UCHT1:

49. The binding partner of any one of claims 45-48, wherein the binding partner comprises a sequence at least 90% similar to any of the following sequences excluding the underlined sequences:

50. The binding partner of any one of claims 45-48, wherein the binding partner is a single chain diabody (scDb) and comprises a sequence at least 90% identical to any of the following sequences excluding the underlined sequences:

51. The binding partner of any one of claims 1-48, wherein the binding partner comprises a heavy chain constant region selected from the group consisting of human IgM, IgGl, IgG2, IgG3, IgG4, IgA1 and IgA2.

52. The binding partner of claim 51, wherein the heavy chain constant region comprises one or more amino acid substitutions in the Fc region.

53. A binding partner according to any one of claims 1-48, 51 or 52, wherein the binding partner comprises a human kappa light chain constant region or a human lambda light chain constant region.

54. The binding partner of any one of claims 45-50, wherein the binding partner is in the form of a CrossMab.

55. The binding partner of any one of claims 1-54, wherein the binding partner is conjugated to a detectable label, a chemotherapeutic agent, a radioisotope, or a toxin.

56. The binding partner of any one of claims 1-54, wherein the binding partner is contained within a chimeric antigen receptor.

57. The binding partner of claim 56, wherein the binding partner is expressed by a T cell, a macrophage, a neutrophil or a natural killer cell.

58. A binding partner according to any of the preceding claims, wherein binding of the binding partner to the peptide conjugate / MHC complex is not inhibited by a free targeted covalent inhibitor.

59. A complex comprising a binding partner according to any one of claims 1-58 and a peptide conjugate / MHC complex.

60. A polynucleotide encoding a binding partner according to any one of claims 1-58.

61. A polynucleotide encoding the heavy chain variable region and / or light chain variable region of a binding partner according to any one of claims 1-58.

62. A vector comprising the polynucleotide according to claim 60 or 61.

63. The vector of claim 62, wherein the vector is a viral vector.

64. The vector of claim 63, wherein the viral vector is an adenoviral vector, a lentiviral vector, a retroviral vector, or an adeno-associated viral vector.

65. A recombinant host cell, comprising: (a) a polynucleotide according to claim 60 or 61; (b) a vector according to any one of claims 62 to 64; (c) a first polynucleotide encoding the VH or heavy chain of the binding partner according to any one of claims 1-58, and a second polynucleotide encoding the VL or light chain of the binding partner according to any one of claims 1-58; or (d) a first vector comprising a first polynucleotide encoding the VH or heavy chain of the binding partner according to any one of claims 1-58, and a second vector comprising a second polynucleotide encoding the VL or light chain of the binding partner according to any one of claims 1-58.

66. A pharmaceutical composition comprising a binding partner according to any one of claims 1-58, a polynucleotide according to claim 60 or 61, a vector according to any one of claims 62-64 or a host cell according to claim 65, and a pharmaceutically acceptable carrier or excipient.

67. A method of producing a binding partner, the method comprising culturing the host cell according to claim 65 under suitable conditions such that the polynucleotide is expressed and the binding partner is produced.

68. A eukaryotic cell comprising a polynucleotide according to claim 60 or 61 or a vector according to any one of claims 62-64, wherein the cell is optionally a totipotent, pluripotent or multipotent 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 γδT cell, optionally a natural killer cell, a natural killer T cell, a mucosa-associated invariant T (MAIT) cell, a neutrophil or a macrophage.

69. A method comprising administering to an individual in need thereof a binding partner according to any one of claims 1-58, a polynucleotide according to claim 60 or 61, a vector according to any one of claims 62-64, a pharmaceutical composition according to claim 66, or a cell according to claim 68.

70. A method for producing a peptide conjugate / MHC complex, the method comprising contacting a cell with a targeted covalent inhibitor, and isolating the peptide conjugate / MHC complex.

71. The method of claim 70, further comprising identifying the peptide conjugate / MHC complex.

72. A cell-free peptide conjugate / MHC complex comprising: (a) an isolated peptide conjugate formed by a covalent reaction of a targeted covalent inhibitor with a peptide; and (b) MHC.

73. The peptide conjugate / MHC complex of claim 72, wherein RAS, EGFR, BTK, HER2 / NEU (ERBB2), HER3 (ERBB3), HER4 (ERBB4), MET (HGFR); FGFR, CDK, acetylcholinesterase (ACHE), p90 ribosomal S6 kinase (RSK), TP53, IDH1, GNAS, FBXW7, CTNNB1, DNMT3A, cathepsin B, cathepsin C, cathepsin F, cathepsin H, cathepsin K, cathepsin L, cathepsin O, cathepsin S, cathepsin V, cathepsin W, cathepsin X, caspase, pancreatic lipase, METAP2, any cancer testis antigen (CTA), long interspersed element-1 (LINE-1), short interspersed elements optionally being Alu, or any endogenous retroviral protein, and optionally wherein the RAS is KRAS, HRAS or NRAS.

74. The antigen of claim 72 or 73, wherein the targeted covalent inhibitor is osimertinib, ibrutinib, neratinib, sotolacib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1 or BI 182391.

75. A cell-free peptide conjugate / MHC complex comprising (a) a compound selected from the group consisting of compounds 1 and 4-8, which is covalently bound to a cysteine ​​residue in a peptide comprising the amino acid sequence of VVVGACGVGK, VVGACGVGK or KLVVVGACGV; and (b) MHC.

76. A cell-free peptide conjugate / MHC complex comprising (a) Compound 2: which is covalently bound to a cysteine ​​residue in a peptide comprising the amino acid sequence of QLMPFGCLL, LMPFGCLLDY or MFPGCLLDY; and (b) MHC.

77. A cell-free peptide conjugate / MHC complex comprising (a) Compound 3: which is covalently bound to a cysteine ​​residue in a peptide comprising the amino acid sequence of YMANGCLLNY; and (b) MHC.

78. The peptide conjugate / MHC complex of any one of claims 72-77, wherein the MHC is HLA, optionally wherein the HLA is HLA-A*02:01, HLA-A*03:01, HLA-A*01:01, HLA-A*11:01, HLA-A*24:02, HLA-A*26:01, HLA-B*07:02, HLA-B*08:01, HLA-B*27:05, HLA-B*39:01, HLA-B*40:01, HLA-B*58:01 or HLA-B*15:

01.

79. A recombinant cell or particle comprising on its outer surface the cell-free peptide conjugate / MHC complex according to any one of claims 72-78.

80. A method for identifying one or more binding partners that specifically bind to a peptide conjugate presented by HLA, the method comprising contacting the cell-free peptide conjugate / MHC complex according to any one of claims 72-78 with a plurality of binding partners and selecting one or more binding partners that specifically bind to the cell-free peptide conjugate / MHC complex.

81. The method of claim 80, further comprising determining the sequence of the one or more binding partners selected.

82. The method of claim 80 or 81, further comprising producing the selected one or more binding partners.

83. A method for identifying binding partners that specifically bind to a peptide conjugate presented by two or more HLAs, the method comprising providing a cell sample from a subject who has been treated with a targeted covalent inhibitor and has different HLA types, or providing a cell-free peptide conjugate / MHC complex according to any one of claims 72-78 presented by two or more HLAs, and screening binding partners to identify binding partners that specifically bind to a peptide conjugate or antigen presented by more than one HLA.

84. The method of claim 74, wherein the two or more HLA types include HLA-A*02:01 and at least one additional HLA type.

85. The method of claim 74 or 75, wherein the two or more HLAs are each HLA-A*02:01, HLA-A*03:01, HLA-A*01:01, HLA-A*11:01, HLA-A*24:02, HLA-A*26:01, HLA-B*07:02, HLA-B*08:01, HLA-B*27:05, HLA-B*39:01, HLA-B*40:01, HLA-B*58:01 or HLA-B*15:

01.

86. A method of killing cancer cells in a subject, the method comprising administering to the subject: (a) targeted covalent inhibitors, and (b) A binding partner according to any one of claims 1 to 58, a polynucleotide according to claim 60 or 61, a vector according to any one of claims 62 to 64, a pharmaceutical composition according to claim 66, or a cell according to claim 68.

87. The method of claim 86, wherein the targeted covalent inhibitor targets RAS, Bruton's tyrosine kinase (BTK), EGFR (ERBB1), HER2 / NEU (ERBB2), HER3 (ERBB3), HER4 (ERBB4), fibroblast growth factor receptor (FGFR), MET, BRAF, cyclin-dependent kinase (CDK), acetylcholinesterase (ACHE), TP53, IDH1, GNAS, FBXW7, CTNNB1, DNMT3A, cathepsins, caspases, pancreatic lipase, METAP2, any cancer testis antigen, endogenous retroviral protein, long interspersed element-1 (LINE-1) or short interspersed element (SINE), and optionally wherein the RAS is KRAS, HRAS or NRAS.

88. A method of targeting cells expressing EGFR, BTK or RAS mutations in a subject treated with an EGFR, BTK or RAS targeted covalent inhibitor, the method comprising administering to the subject a binding partner according to any one of claims 1-58, a polynucleotide according to claim 60 or 61, a vector according to any one of claims 62-64, a pharmaceutical composition according to claim 66, or a cell according to claim 68.

89. The method of claim 88, wherein the subject has cancer.

90. A method of treating cancer in a subject who has been treated with a targeted covalent inhibitor, the method comprising administering to the subject a binding partner according to any one of claims 1-58, a polynucleotide according to claim 60 or 61, a vector according to any one of claims 62-64, a pharmaceutical composition according to claim 66, or a cell according to claim 68.

91. A method of treating a disease or disorder in a subject who has been treated with a targeted covalent inhibitor, the method comprising administering to the subject a binding partner according to any one of claims 1-58, a polynucleotide according to claim 60 or 61, a vector according to any one of claims 62-64, a pharmaceutical composition according to claim 66, or a cell according to claim 68.

92. The method of claim 91, wherein the disease or disorder is an autoimmune disease or a fibrotic disease.

93. A method of enhancing immune recognition of cells expressing RAS or EGFR mutations in a subject having a cancer exhibiting RAS or EGFR mutations, the method comprising administering to the subject: (a) RAS or EGFR inhibitors, and (b) the binding partner of any one of claims 1 to 58, the polynucleotide of claims 60 or 61, the vector of any one of claims 62 to 64, the pharmaceutical composition of claim 66 or the cell of claim 68, and optionally wherein the RAS mutation is KRAS G12C 、KRAS G12D 、KRAS G12R 、KRAS G12S 、HRAS G12C 、HRAS G12D 、HRAS G12R 、HRAS G12S NRAS G12C NRAS G12D NRAS G12R or NRAS G12S .

94. The method of claim 93, wherein the subject has been previously treated with a RAS or EGFR inhibitor.

95. The method of claim 93 or 94, wherein the inhibitor is osimertinib, ibrutinib, neratinib, sotolacib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1, or BI 182391.

96. The method of any one of claims 90-92, wherein the targeted covalent inhibitor targets RAS, Bruton's tyrosine kinase (BTK), EGFR (ERBB1), HER2 / NEU (ERBB2), HER3 (ERBB3), HER4 (ERBB4), fibroblast growth factor receptor (FGFR), MET, BRAF, cyclin-dependent kinase (CDK), acetylcholinesterase (ACHE), TP53, IDH1, GNAS, FBXW7, CTNNB1, DNMT3A, cathepsins, caspases, pancreatic lipase, METAP2, any cancer testis antigen, endogenous retroviral protein, long interspersed element-1 (LINE-1) or short interspersed element (SINE).

97. The method of any one of claims 86, 87, 89, 90, or 93-96, wherein the cancer is renal cell carcinoma, breast cancer, prostate cancer, pancreatic cancer, lung cancer, liver cancer, ovarian cancer, cervical cancer, colorectal cancer, esophageal cancer, glioma, glioblastoma, brain cancer, gastric cancer, bladder cancer, testicular cancer, head and neck cancer, melanoma, skin cancer, sarcoma, fibrosarcoma, angiosarcoma, osteosarcoma, rhabdomyosarcoma, leukemia, lymphoma, or myeloma.

98. The method of any one of claims 86-90, wherein the targeted covalent inhibitor is osimertinib, ibrutinib, neratinib, sotolacib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1, or BI 182391.

99. The method of any one of claims 86-98, further comprising administering an additional therapeutic agent.

100. The method of claim 99, wherein the additional therapeutic agent is a chemotherapy, an immunomodulator, or another targeted covalent inhibitor.

101. The method of claim 100, wherein 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-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-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 anchored IL2 or engineered IL2; or an extracellular adenosine (eADO) signaling inhibitor.

102. A method for detecting a peptide conjugate / MHC complex in a biological sample, the method comprising contacting the sample with a binding partner according to any one of claims 1-58, wherein the peptide conjugate / MHC complex comprises a peptide conjugate formed by a covalent reaction of a targeted covalent inhibitor and a peptide.

103. The method of claim 102, wherein the peptide is an EGFR, BTK or RAS peptide, and optionally wherein the RAS peptide is KRAS G12C Peptide, KRAS G12D Peptide, KRAS G12R Peptide, KRAS G12S Peptide, HRAS G12C Peptide, HRAS G12D Peptide, HRAS G12R Peptide, HRAS G12S Peptide, NRAS G12C Peptide, NRAS G12D Peptide, NRAS G12R Peptide, or NRAS G12S Peptide.

104. The method of claim 102 or 103, wherein the targeted covalent inhibitor is EGFR, BTK or KRAS G12C Inhibitor.

105. The method of any one of claims 102-104, wherein the targeted covalent inhibitor is osimertinib, ibrutinib, neratinib, sotolacib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1, or BI 182391.

106. The method of any one of claims 102-105, wherein the biological sample is blood or serum.

107. A method of identifying a cell comprising a peptide conjugate / MHC complex according to any one of claims 72-78, the method comprising contacting the cell with a binding partner according to any one of claims 1-58.

108. A method for producing one or more binding partners that specifically bind to a peptide conjugate, wherein the peptide conjugate comprises a peptide covalently bound to a non-peptide molecule, the method comprising: a) exposing the peptide conjugate to a plurality of binding partners, and b) selecting a binding partner that specifically binds to the peptide conjugate to provide the selected one or more binding partners.

109. The method of claim 108, wherein step (b) comprises selecting a binding partner that specifically binds to the peptide conjugate but does not detectably bind to or binds with a lower affinity to the peptide or the non-peptide molecule when the peptide and the non-peptide molecule are not covalently bound.

110. A method for producing one or more binding partners that specifically bind to a peptide conjugate presented in the context of an MHC molecule or a fragment or derivative thereof, wherein the peptide conjugate comprises a peptide covalently bound to a non-peptide molecule, the method comprising: a) providing a complex of the peptide conjugate and the MHC molecule or a fragment or derivative thereof; b) exposing the complex to a plurality of binding partners, and c) selecting a binding partner that specifically binds to the complex to provide the selected one or more binding partners.

111. The method of claim 110, wherein step (c) comprises selecting a binding partner that specifically binds to the complex but does not detectably bind to or binds with a lower affinity to the complex of the peptide and the MHC molecule or a fragment thereof, wherein the peptide is not covalently bound to the non-peptide molecule.

112. The method of claim 110 or claim 111, wherein step (c) comprises selecting a binding partner that specifically binds to the peptide conjugate presented in the context of two or more different MHC molecules.

113. The method according to any one of claims 110-112, wherein the MHC molecule is an MHC class I molecule and the peptide is 7-15 amino acids in length, or wherein the MHC molecule is a non-classical MHC class I molecule and the peptide is 7-15 amino acids in length.

114. The method according to any one of claims 110-112, wherein the MHC molecule is an MHC class II molecule and the peptide is 9-30 amino acids in length.

115. The method of any one of claims 110-114, wherein the complex is immobilized on a solid support.

116. The method of any one of claims 110-114, wherein the complex is presented on the surface of a cell.

117. The method of claim 116, further comprising determining whether the one or more binding partners selected in step (c) are capable of mediating immune cell-mediated killing or antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) or complement-dependent cytotoxicity (CDC)-mediated killing of the cells, or by using the binding partner as an immunotoxin or as an antibody-drug conjugate (ADC) or as a radioactive conjugate.

118. The method of any one of claims 110-117, wherein the plurality of binding partners is produced by a phage display library or a yeast display library.

119. A method for killing cancer cells, the method comprising administering to the cells a binding partner that specifically binds to a peptide conjugate, wherein the peptide conjugate comprises (i) a peptide derived from a target protein within the cell, the peptide being covalently bound to (ii) a non-peptide molecule, and wherein the binding partner mediates immune cell-mediated killing of the cells or antibody-drug conjugate (ADC)-mediated, antibody-dependent cellular phagocytosis (ADCP) or complement-dependent cytotoxicity (CDC) killing of the cells, or by using the binding partner as an immunotoxin, or by the action of a radioactive conjugate, thereby killing the cancer cells.

120. The method of claim 119, wherein the peptide conjugate is presented on the cell in the context of an MHC molecule.

121. The method of claim 120, wherein the binding partner specifically binds to a complex comprising the peptide conjugate and the MHC molecule.

122. The method of claim 121, wherein the binding partner undetectably binds to a complex of the peptide and the MHC molecule, wherein the peptide is not covalently bound to the non-peptide molecule.

123. The method of any one of claims 119-122, further comprising administering the non-peptide molecule to the cell, wherein the non-peptide molecule forms a covalent bond with the target protein in the cell.

124. The method of claim 123, wherein the non-peptide molecule is administered to the cells prior to administration of the binding partner.

125. The method of any one of claims 119-124, wherein the cell is isolated from a subject.

126. The method of any one of claims 119-125, wherein the cell is in a subject and the binding partner is administered to the subject.

127. The method of claim 123 or claim 124, wherein the cell is in a subject and the non-peptide molecule and the binding partner are administered to the subject.

128. The method of any one of claims 119-127, wherein the target protein is alternatively spliced ​​or overexpressed in cancer cells but is not alternatively spliced ​​or overexpressed by cancer cells.

129. The method of claim 128, wherein the target protein is encoded by a gene that is mutated in cancer cells but not in non-cancerous cells.

130. The method of claim 128 or claim 129, wherein the non-peptide molecule is a covalent inhibitor of the target protein.

131. The method of any one of claims 119-131, wherein the binding partner is a complete antibody, a bispecific antibody, a multispecific antibody, an antigen binding (Fab) fragment, a Fab' fragment, a (Fab')2 fragment, Fd, Fv, dAb, a single domain fragment or a single monomer variable antibody domain, a single chain diabody (scDb), a diabody (Db), a dual affinity retargeting (DART) molecule, a single chain variable fragment (scFv), a bispecific T cell engaging antibody (BiTE), a bispecific killer cell engaging antibody (BiKE), a CrossMab, a camelid antibody, a trispecific binding partner, a chimeric antigen receptor (CAR), a monomer (also known as Adnectin), a DARPin, an anticalculin, an affibody or an antibody-mimicking protein, or comprises a radioconjugate.

132. The method of claim 131, wherein the CAR is present on a T cell, a natural killer (NK) cell, a neutrophil, or a macrophage.

133. The method of any one of claims 119-130, wherein the binding partner is an antibody-drug conjugate (ADC), a radioconjugate, or a toxin conjugate.

134. The method of any one of claims 119-133, wherein the target protein is selected from KRAS, Bruton's tyrosine kinase (BTK), epidermal growth factor receptor (EGFR) family members, fibroblast growth factor receptor (FGFR), MET, BRAF, cyclin-dependent kinase (CDK), acetylcholinesterase (ACHE), TP53, IDH1, GNAS, FBXW7, CTNNB1, DNMT3A, cathepsin, caspase, pancreatic lipase, METAP2, cancer testis antigen, viral polymerase, protein required for viral cell entry, protein encoded by a transposable element (e.g., endogenous retrovirus), or a mutant thereof.

135. The method of claim 134, wherein the target protein is a KRAS protein comprising a G12C mutation, and the non-peptide molecule is selected from sotolacib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, RMC-6291, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1 or BI 182391 and derivatives thereof.

136. The method of claim 134, wherein the target protein is EGFR, and the non-peptide molecule is selected from PD168393, PF00299804 (dacomitinib), EKB569 (pelitinib), afatinib, WZ4002, osimertinib (AZD9291), PF-06459988, nazatinib, naquintinib, omotinib, avitinib, rociletinib, neratinib, pyrotinib, poziotinib and derivatives thereof.

137. The method of claim 134, wherein the target protein is Bruton's tyrosine kinase (BTK) and the non-peptide molecule is selected from ibrutinib, acalabrutinib, zanubrutinib, CHMFL-BTK-11, ONO / GS-405, PRN1008, CC-292 and derivatives thereof.

138. The method of claim 134, wherein the target protein is p90 ribosomal S6 kinase (RSK) and the non-peptide molecule is fluoromethyl ketone (FMK), dimethyl fumarate or a derivative thereof.

139. The method of claim 134, wherein the target protein is FGFR and the non-peptide molecule is selected from FIIN-1, FIIN-2, FIIN-3, BGJ398, AZD4547, PRN1371, FGF401 and derivatives thereof.

140. A method for treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of a binding partner that specifically binds to a peptide conjugate, wherein the peptide conjugate comprises (i) a peptide derived from a target protein present in a cancer cell of the subject, the peptide being covalently bound to (ii) a non-peptide molecule, and wherein the binding partner mediates immune cell-mediated killing or antibody-drug conjugate (ADC)-mediated killing of the cancer cell, antibody-dependent cellular phagocytosis (ADCP) or complement-dependent cytotoxicity (CDC)-mediated killing of the cancer cell, or by using the binding partner as an immunotoxin or radioconjugate in the subject, thereby killing the cancer cell.

141. The method of claim 140, wherein the peptide conjugate is presented on the subject's cancer cells in the context of an MHC molecule.

142. The method of claim 140, wherein the binding partner specifically binds to a complex comprising the peptide conjugate and the MHC molecule.

143. The method of claim 142, wherein the binding partner undetectably binds to a complex of the peptide and the MHC molecule, wherein the peptide is not covalently bound to the non-peptide molecule.

144. The method of any one of claims 140-143, wherein the subject has previously received the non-peptide molecule.

145. The method of any one of claims 140-143, further comprising administering to the subject an effective amount of the non-peptide molecule, wherein the non-peptide molecule forms a covalent bond with the target protein in the cancer cells of the subject.

146. The method of claim 145, wherein the non-peptide molecule is administered to the subject prior to administration of the binding partner.

147. A method for improving the efficacy of an anti-cancer therapy in a subject in need thereof, wherein the anti-cancer therapy comprises administering to the subject a non-peptide molecule that forms a covalent bond with a target protein in the subject's cancer cells, the method further comprising administering to the subject an effective amount of a binding partner that specifically binds to a peptide conjugate, wherein the peptide conjugate comprises a peptide derived from the target protein covalently bound to the non-peptide molecule, and wherein the binding partner mediates immune cell-mediated killing or antibody-drug conjugate (ADC)-mediated killing of the cancer cells, antibody-dependent cellular phagocytosis (ADCP) or complement-dependent cytotoxicity (CDC)-mediated killing of the cancer cells, or by using the binding partner as an immunotoxin or by using the binding partner as a radioconjugate in the subject.

148. The method of claim 147, wherein the peptide conjugate is presented on the subject's cancer cells in the context of an MHC molecule.

149. The method of claim 148, wherein the binding partner specifically binds to a complex comprising the peptide conjugate and the MHC molecule.

150. The method of claim 149, wherein the binding partner undetectably binds to a complex of the peptide and the MHC molecule, wherein the peptide is not covalently bound to the non-peptide molecule.

151. The method of any one of claims 147-149, wherein the subject has received the non-peptide molecule prior to administration of the binding partner.

152. A kit comprising: (i) a non-peptide molecule, wherein the 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 the peptide conjugate comprises a peptide derived from the target protein covalently bound to the non-peptide molecule, and optionally (iii) instructions for use.

153. An isolated peptide conjugate comprising a peptide of 7-30 amino acids in length, said peptide comprising an amino acid sequence at least 80% identical to the amino acid sequence VVGACGVGK, wherein said peptide is conjugated to sotolacib or a derivative thereof.

154. An isolated peptide conjugate comprising a peptide of 7-30 amino acids in length, said peptide comprising an amino acid sequence at least 80% identical to the amino acid sequence QLMPFGCLL, wherein said peptide is conjugated to osimertinib or a derivative thereof.

155. An isolated peptide conjugate comprising a peptide of 7-30 amino acids in length, said peptide comprising an amino acid sequence at least 80% identical to the amino acid sequence YMANGCLLNY, wherein said peptide is conjugated to ibrutinib or a derivative thereof.

156. An isolated molecular complex comprising a peptide conjugate according to any one of claims 46-48 and an MHC molecule or a fragment or derivative thereof.

157. A host cell comprising the molecular complex according to claim 156.

158. A solid surface support comprising the molecular complex according to claim 156.

159. A kit comprising (i) a peptide conjugate according to any one of claims 153-155, a molecular complex according to claim 156, a cell according to claim 157, a solid surface support according to claim 158, or any combination thereof, and optionally (ii) instructions for use.

160. An isolated binding partner that specifically binds to the peptide conjugate of any one of claims 46-48 or the molecular complex of claim 156.

161. The isolated binding partner of claim 160, wherein the binding partner is an antibody or an antigen-binding fragment thereof.

162. An isolated binding partner according to claim 161, wherein the binding partner is a component of a chimeric antigen receptor (CAR).

163. A fusion protein comprising a binding partner according to any of the preceding claims, wherein the fusion protein comprises (i) at least one cytokine, and / or (ii) an additional antibody or fragment thereof that is an immune checkpoint inhibitor.

164. A binding partner according to claim 163, wherein (i) the cytokine is selected from the group consisting of interleukin (IL) -2, IL-7, IL-8, IL-15, IL-17 and combinations thereof, and / or (ii) the additional antibody or fragment thereof that is an immune checkpoint inhibitor is an 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-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 or an agonist anti-CD28 antibody.

165. A binding partner according to any of the preceding claims, wherein the binding partner comprises a component that binds to a T cell or natural killer (NK) cell protein, wherein the T cell or NK cell protein is selected from the group consisting of T cell receptor proteins, CD4, CD8, CD28, CD16A, NKG2D, NKp30, NKp46 and combinations thereof.

166. 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 heavy chain complementary determining region 3 (CDR-H3), comprising an amino acid sequence of GX1WX2X3AMDY, wherein X1 is G, R, H, S or K, X2 is Y or I, and X3 is P or A; and / or (b) the VL comprises: i. Light chain complementary determining region 3 CDR-L3, comprising an amino acid sequence of QQX1SYVX2X3X4IT, wherein X1 is I, A, P, V or S, X2 is K, R, A or H, X3 is K or R, and X4 is L, T, K, R, V, A or E. i. Light chain complementary determining region 3 CDR-L3, comprising an amino acid sequence of QQX1SYVX2X3X4IT, wherein X1 is I, A, P, V or S, X2 is K, R, A or H, X3 is K or R, and X4 is L, T, K, R, V, A or E.

167. 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 a fragment thereof; and wherein the polypeptide binds to an epitope of MHC.

168. 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 a fragment thereof; and wherein the dissociation constant (K) of the antigen binding domain binding to the peptide conjugate / MHC complex is D ) is at most about 50 nM.

169. 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 a fragment thereof; and wherein the dissociation constant (K) of the antigen binding domain for binding to a free targeted covalent inhibitor is D ) is at least 200 nM.

170. 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 a fragment thereof; and wherein: (a) the polypeptide is bound to the peptide conjugate / MHC complex at an angle of about 10° to 60° between the axis of the MHC and the axis of the polypeptide; (b) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is about 10° to 60°; (c) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is different from the angle between the axis of the TCR and the axis of the peptide conjugate / MHC complex when the TCR is bound to the peptide conjugate / MHC complex; (d) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is about 10° to 60° or about 10° to 60° less than the angle between the axis of the TCR and the axis of the peptide conjugate / MHC complex when the TCR is bound to the peptide conjugate / MHC complex; (e) the polypeptide binds to the peptide conjugate / MHC complex at an angle that is different from the angle at which the T cell receptor binds to the peptide conjugate / MHC complex; and / or (f) the polypeptide binds to the peptide conjugate / MHC complex at an angle of about 10° to 60° or about 10° to 60° less than the angle at which a T cell receptor binds to the peptide conjugate / MHC complex.

171. 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 a fragment thereof; and wherein the polypeptide contacts one or more residues of the α1 domain or region and one or more residues of the α2 domain or region of the heavy chain of the MHC of the peptide conjugate / MHC complex.

172. 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 a fragment thereof; and wherein the polypeptide binds to residues 62-66, 106-109 and / or 150-170 of MHC or one or more residues thereof.

173. 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 a fragment thereof; and wherein the interface area of ​​the polypeptide with the peptide conjugate / MHC complex is at least about 174. A polypeptide according to any one of claims 166-173, wherein (a) the VH comprises a CDR-H3 comprising an amino acid sequence of GX1WX2X3AMDY, wherein X1 is G, R, H, S or K, X2 is Y or I, and X3 is P or A; and / or (b) the VL comprises a CDR-L3 comprising an amino acid sequence of QQX1SYVX2X3X4IT, wherein X1 is I, A, P, V or S, X2 is K, R, A or H, X3 is K or R, and X4 is L, T, K, R, V, A or E.

175. The polypeptide of any one of claims 166-174, wherein the VH comprises: (i) CDR-H1, which comprises the amino acid sequence of DYSIH, or a variant thereof comprising 1-3 amino acid changes; (ii) CDR-H2 comprising the amino acid sequence of SISSSSGSTSYADSVKG, or a variant thereof comprising 1-5 amino acid changes; and / or (iii) CDR-H3, comprising an amino acid sequence of GX1WX2X3AMDY, wherein X1 is G, R, H, S or K, X2 is Y or I, and X3 is P or A.

176. The polypeptide of any one of claims 166-175, wherein the VL comprises: (i) CDR-L1 comprising the amino acid sequence of RASQSVSSAVA, or a variant thereof comprising 1-5 amino acid changes; (ii) CDR-L2 comprising the amino acid sequence of SASSLYS, or a variant thereof comprising 1-5 amino acid changes, and / or (iii) CDR-L3, which comprises the amino acid sequence of QQX1SYVX2X3X4IT, wherein X1 is I, A, P, V or S, X2 is K, R, A or H, X3 is K or R, and X4 is L, T, K, R, V, A or E.

177. The polypeptide of claim 166, wherein the VH comprises a CDR-H1 having the sequence of DYSIH, a CDR-H2 having the sequence of SISSSSGSTSYADSVKG, and a CDR-H3 having the sequence of GGWIAAMDY.

178. The polypeptide of claim 166 or 177, wherein the VL comprises a CDR-L1 having the sequence of RASQSVSSAVA, a CDR-L2 having the sequence of SASSLYS, and a CDR-L3 having the sequence of QQASYVRKTIT.

179. A polypeptide according to any one of claims 166-178, wherein the VH comprises an amino acid sequence having at least about 90%, 95% or 100% sequence identity to the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYSIHWVRQAPGKGLEWVASISSSSGSTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGWIAAMDYWGQGTLVTVSS.

180. The polypeptide of any one of claims 166-179, wherein the VL comprises an amino acid sequence having at least about 90%, 95% or 100% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

181. A polypeptide according to any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GSWIHAMDY.

182. The polypeptide of claim 181, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SISSSWGVTSYADSVKG.

183. The polypeptide of claim 181 or 182, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FHWYSIH.

184. The polypeptide of any one of claims 181-183, wherein the antigen binding domain further comprises a light chain complementary determining region 3 (VL), and wherein the VL comprises a CDR-L3 comprising the amino acid sequence of QQASYVRKTIT.

185. The polypeptide of claim 184, wherein the VL comprises a CDR-L2 comprising the amino acid sequence of SASSLYS.

186. The polypeptide of claim 184 or 185, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

187. The polypeptide of any one of claims 181-186, wherein the antigen binding domain comprises: CDR-H3 sequence of GSWIHAMDY, The CDR-H2 sequence of SISSSWGVTSYADSVKG, CDR-H1 sequence of FHWYSIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

188. The polypeptide of any one of claims 181-187, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFHWYSIHWVRQAPGKGLEWVASISSSWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIHAMDYWGQGTLVTVSS.

189. The polypeptide of any one of claims 184-187, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

190. The polypeptide of any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementarity determining region 3 (CDR-H3), and the heavy chain complementarity determining region 3 (CDR-H3) comprises the amino acid sequence of GHWIAAMDY.

191. The polypeptide of claim 190, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SIASSSGSTGYADSVKG.

192. The polypeptide of claim 190 or 191, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FSWYSIH.

193. A polypeptide according to any one of claims 190-192, wherein the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises a light chain complementary determining region 3 (CDR-L3), and the light chain complementary determining region 3 (CDR-L3) comprises the amino acid sequence of QQASYVRKTIT.

194. A polypeptide according to claim 193, wherein the VL comprises a CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS.

195. The polypeptide of claim 193 or 194, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

196. The polypeptide of any one of claims 190-195, wherein the antigen binding domain comprises: CDR-H3 sequence of GHWIAAMDY, The CDR-H2 sequence of SIASSSGSTGYADSVKG, CDR-H1 sequence of FSWYSIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

197. A polypeptide according to any one of claims 190-196, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSWYSIHWVRQAPGKGLEWVASIASSSGSTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGHWIAAMDYWGQGTLVTVSS.

198. A polypeptide according to any one of claims 193-196, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

199. A polypeptide according to any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GGVIHAMDY.

200. The polypeptide of claim 199, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SILSRWGVTSYADSVKG.

201. The polypeptide of claim 199 or 200, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FSPYSIH.

202. The polypeptide of any one of claims 199-201, wherein the antigen binding domain further comprises a light chain complementary determining region 3 (VL), and wherein the VL comprises a CDR-L3 comprising the amino acid sequence of QQASYVRKTIT.

203. The polypeptide of claim 202, wherein the VL comprises a CDR-L2 comprising the amino acid sequence of SASSLYS.

204. The polypeptide of claim 202 or 203, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

205. The polypeptide of any one of claims 199-204, wherein the antigen binding domain comprises: CDR-H3 sequence of GGVIHAMDY, The CDR-H2 sequence of SILSRWGVTSYADSVKG, CDR-H1 sequence of FSPYSIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

206. The polypeptide of any one of claims 199-205, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYSIHWVRQAPGKGLEWVASILSRWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGVIHAMDYWGQGTLVTVSS.

207. The polypeptide of any one of claims 202-205, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

208. A polypeptide according to any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementarity determining region 3 (CDR-H3), and the heavy chain complementarity determining region 3 (CDR-H3) comprises the amino acid sequence of GSWIAAMDY.

209. The polypeptide of claim 208, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SISSWHGETGYADSVKG.

210. The polypeptide of claim 208 or 209, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FSPYSIH.

211. A polypeptide according to any one of claims 208-210, wherein the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises a CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT.

212. The polypeptide of claim 211, wherein the VL comprises a CDR-L2 comprising the amino acid sequence of SASSLYS.

213. The polypeptide of claim 211 or 212, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

214. The polypeptide of any one of claims 208-213, wherein the antigen binding domain comprises: CDR-H3 sequence of GSWIAAMDY, The CDR-H2 sequence of SISSWHGETGYADSVKG, CDR-H1 sequence of FSPYSIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

215. The polypeptide of any one of claims 208-214, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYSIHWVRQAPGKGLEWVASISSWHGETGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS.

216. The polypeptide of any one of claims 211-214, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

217. A polypeptide according to any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GGWIAAMDY.

218. The polypeptide of claim 217, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SISSLQGDTGYADSVKG.

219. The polypeptide of claim 217 or 218, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FSWYSIH.

220. The polypeptide of any one of claims 217-219, wherein the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises a CDR-L3 comprising the amino acid sequence of QQASYVRKTIT.

221. A polypeptide according to claim 220, wherein the VL comprises a CDR-L2, and the CDR-L2 comprises the amino acid sequence of SASSLYS.

222. The polypeptide of claim 220 or 221, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

223. The polypeptide of any one of claims 217-222, wherein the antigen binding domain comprises: CDR-H3 sequence of GGWIAAMDY, The CDR-H2 sequence of SISSLQGDTGYADSVKG, CDR-H1 sequence of FSWYSIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

224. The polypeptide of any one of claims 217-223, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSWYSIHWVRQAPGKGLEWVASISSLQGDTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGWIAAMDYWGQGTLVTVSS.

225. The polypeptide of any one of claims 220-223, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

226. A polypeptide according to any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GSWIAAMDY.

227. The polypeptide of claim 226, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SIASWYGDTGYADSVKG.

228. The polypeptide of claim 226 or 227, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FHYYSIH.

229. The polypeptide of any one of claims 226-228, wherein the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises a CDR-L3 comprising the amino acid sequence of QQASYVRKTIT.

230. The polypeptide of claim 229, wherein the VL comprises a CDR-L2 comprising the amino acid sequence of SASSLYS.

231. The polypeptide of claim 229 or 230, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

232. The polypeptide of any one of claims 229-231, wherein the antigen binding domain comprises: CDR-H3 sequence of GSWIAAMDY, The CDR-H2 sequence of SIASWYGDTGYADSVKG, The CDR-H1 sequence of FHYYSIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

233. The polypeptide of any one of claims 226-232, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFHYYSIHWVRQAPGKGLEWVASIASWYGDTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS.

234. A polypeptide according to any one of claims 229-232, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

235. A polypeptide according to any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GGRIEAMDY.

236. The polypeptide of claim 235, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SISSWYGKTGYADSVKG.

237. The polypeptide of claim 235 or 236, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FGYYSIH.

238. A polypeptide according to any one of claims 235-237, wherein the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises a CDR-L3, and the CDR-L3 comprises the amino acid sequence of QQASYVRKTIT.

239. The polypeptide of claim 238, wherein the VL comprises a CDR-L2 comprising the amino acid sequence of SASSLYS.

240. The polypeptide of claim 238 or 239, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

241. The polypeptide of any one of claims 238-240, wherein the antigen binding domain comprises: CDR-H3 sequence of GGRIEAMDY, The CDR-H2 sequence of SISSWYGKTGYADSVKG, The CDR-H1 sequence of FGYYSIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

242. The polypeptide of any one of claims 236-241, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFGYYSIHWVRQAPGKGLEWVASISSWYGKTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGRIEAMDYWGQGTLVTVSS.

243. The polypeptide of any one of claims 238-241, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

244. The polypeptide of any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementarity determining region 3 (CDR-H3), and the heavy chain complementarity determining region 3 (CDR-H3) comprises the amino acid sequence of GYWIEAMDY.

245. The polypeptide of claim 244, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SIASSYGSTGYADSVKG.

246. The polypeptide of claim 244 or 245, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FSKYSIH.

247. The polypeptide of any one of claims 244-246, wherein the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises a CDR-L3 comprising the amino acid sequence of QQASYVRKTIT.

248. The polypeptide of claim 247, wherein the VL comprises a CDR-L2 comprising the amino acid sequence of SASSLYS.

249. The polypeptide of claim 247 or 248, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

250. The polypeptide of any one of claims 247-249, wherein the antigen binding domain comprises: CDR-H3 sequence of GYWIEAMDY, The CDR-H2 sequence of SIASSYGSTGYADSVKG, CDR-H1 sequence of FSKYSIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

251. The polypeptide of any one of claims 244-250, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSKYSIHWVRQAPGKGLEWVASIASSYGSTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGYWIEAMDYWGQGTLVTVSS.

252. The polypeptide of any one of claims 247-250, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

253. The polypeptide of any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementarity determining region 3 (CDR-H3), and the heavy chain complementarity determining region 3 (CDR-H3) comprises the amino acid sequence of GSWIAAMDY.

254. The polypeptide of claim 253, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SIHSSIGTTGYADSVKG.

255. The polypeptide of claim 253 or 254, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FGLYSIH.

256. The polypeptide of any one of claims 253-255, wherein the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises a CDR-L3 comprising the amino acid sequence of QQASYVRKTIT.

257. The polypeptide of claim 256, wherein the VL comprises a CDR-L2 comprising the amino acid sequence of SASSLYS.

258. The polypeptide of claim 256 or 257, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

259. The polypeptide of any one of claims 256-258, wherein the antigen binding domain comprises: CDR-H3 sequence of GSWIAAMDY, CDR-H2 sequence of SIHSSIGTTGYADSVKG, CDR-H1 sequence of FGLYSIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

260. The polypeptide of any one of claims 253-259, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFGLYSIHWVRQAPGKGLEWVASIHSSIGTTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS.

261. A polypeptide according to any one of claims 256-259, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

262. A polypeptide according to any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GSVIHAMDY.

263. The polypeptide of claim 262, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SILSWIGKTSYADSVKG.

264. The polypeptide of claim 262 or 263, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FSPYSIH.

265. The polypeptide of any one of claims 262-264, wherein the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises a CDR-L3 comprising the amino acid sequence of QQASYVRKTIT.

266. The polypeptide of claim 265, wherein the VL comprises a CDR-L2 comprising the amino acid sequence of SASSLYS.

267. The polypeptide of claim 265 or 266, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

268. The polypeptide of any one of claims 262-267, wherein the antigen binding domain comprises: CDR-H3 sequence of GSVIHAMDY, The CDR-H2 sequence of SILSWIGKTSYADSVKG, CDR-H1 sequence of FSPYSIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

269. The polypeptide of any one of claims 262-268, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYSIHWVRQAPGKGLEWVASILSWIGKTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSVIHAMDYWGQGTLVTVSS.

270. The polypeptide of any one of claims 265-268, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

271. A polypeptide according to any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GGWIAAMDY.

272. The polypeptide of claim 271, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SIASRWGHTGYADSVKG.

273. The polypeptide of claim 271 or 272, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FSPYHIH.

274. The polypeptide of any one of claims 271-273, wherein the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises a CDR-L3 comprising the amino acid sequence of QQASYVRKTIT.

275. The polypeptide of claim 274, wherein the VL comprises a CDR-L2 comprising the amino acid sequence of SASSLYS.

276. The polypeptide of claim 274 or 275, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

277. The polypeptide of any one of claims 274-276, wherein the antigen binding domain comprises: CDR-H3 sequence of GGWIAAMDY, CDR-H2 sequence of SIASRWGHTGYADSVKG, CDR-H1 sequence of FSPYHIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

278. The polypeptide of any one of claims 271-277, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSPYHIHWVRQAPGKGLEWVASIASRWGHTGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGWIAAMDYWGQGTLVTVSS.

279. A polypeptide according to any one of claims 274-277, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

280. The polypeptide of any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementarity determining region 3 (CDR-H3), and the heavy chain complementarity determining region 3 (CDR-H3) comprises the amino acid sequence of GSWIAAMDY.

281. The polypeptide of claim 280, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SIASLQGITGYADSVKG.

282. The polypeptide of claim 280 or 281, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FHEYSIH.

283. The polypeptide of any one of claims 280-282, wherein the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises a CDR-L3 comprising the amino acid sequence of QQASYVRKTIT.

284. The polypeptide of claim 283, wherein the VL comprises a CDR-L2 comprising the amino acid sequence of SASSLYS.

285. The polypeptide of claim 283 or 284, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

286. The polypeptide of any one of claims 280-285, wherein the antigen binding domain comprises: CDR-H3 sequence of GSWIAAMDY, The CDR-H2 sequence of SIASLQGITGYADSVKG, CDR-H1 sequence of FHEYSIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

287. The polypeptide of any one of claims 280-286, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFHEYSIHWVRQAPGKGLEWVASIASLQGITGYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGSWIAAMDYWGQGTLVTVSS.

288. The polypeptide of any one of claims 283-286, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

289. A polypeptide according to any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GGVIHAMDY.

290. The polypeptide of claim 289, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SILSRWGVTSYADSVKG.

291. The polypeptide of claim 289 or 290, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FSDYSIH.

292. The polypeptide of any one of claims 289-291, wherein the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises a CDR-L3 comprising the amino acid sequence of QQASYVRKTIT.

293. The polypeptide of claim 292, wherein the VL comprises a CDR-L2 comprising the amino acid sequence of SASSLYS.

294. The polypeptide of claim 292 or 293, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

295. The polypeptide of any one of claims 289-294, wherein the antigen binding domain comprises: CDR-H3 sequence of GGVIHAMDY, The CDR-H2 sequence of SILSRWGVTSYADSVKG, The CDR-H1 sequence of FSDYSIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

296. The polypeptide of any one of claims 289-295, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYSIHWVRQAPGKGLEWVASILSRWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGVIHAMDYWGQGTLVTVSS.

297. A polypeptide according to any one of claims 292-295, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

298. A polypeptide according to any one of claims 167-173, wherein the antigen binding domain comprises a heavy chain variable region (VH), and wherein the VH comprises a heavy chain complementary determining region 3 (CDR-H3), and the heavy chain complementary determining region 3 (CDR-H3) comprises the amino acid sequence of GGVIHAMDY.

299. The polypeptide of claim 298, wherein the VH comprises a CDR-H2 comprising the amino acid sequence of SISSRWGVTSYADSVKG.

300. The polypeptide of claim 298 or 299, wherein the VH comprises a CDR-H1 comprising the amino acid sequence of FSDYSIH.

301. The polypeptide of any one of claims 298-300, wherein the antigen binding domain further comprises a light chain variable region (VL), and wherein the VL comprises a CDR-L3 comprising the amino acid sequence of QQASYVRKTIT.

302. The polypeptide of claim 301, wherein the VL comprises a CDR-L2 comprising the amino acid sequence of SASSLYS.

303. The polypeptide of claim 301 or 302, wherein the VL comprises a CDR-L1 comprising the amino acid sequence of RASQSVSSAVA.

304. The polypeptide of any one of claims 298-303, wherein the antigen binding domain comprises: CDR-H3 sequence of GGVIHAMDY, The CDR-H2 sequence of SISSRWGVTSYADSVKG, The CDR-H1 sequence of FSDYSIH, The CDR-L3 sequence of QQASYVRKTIT, The CDR-L2 sequence of SASSLYS, and CDR-L1 sequence of RASQSVSSAVA.

305. The polypeptide of any one of claims 298-304, wherein the VH comprises a sequence having at least 80% sequence identity to the sequence EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYSIHWVRQAPGKGLEWVASISSRWGVTSYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARGGVIHAMDYWGQGTLVTVSS.

306. The polypeptide of any one of claims 301-304, wherein the VL comprises a sequence having at least 80% sequence identity to the sequence DIQMTQSPSSLSASVGDRVTITCRASQSVSSAVAWYQQKPGKAPKLLIYSASSLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQASYVRKTITFGQGTKVEIKRTV.

307. The polypeptide of any one of claims 166-180, wherein the antibody or antigen binding fragment specifically binds to a peptide conjugate / MHC complex, and wherein the antibody or antigen binding fragment interacts with the MHC of the peptide conjugate / MHC complex.

308. The polypeptide of claim 307, wherein 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) MHC.

309. The polypeptide of claim 308, wherein the MHC is human leukocyte antigen (HLA).

310. The polypeptide of claim 309, wherein the HLA is HLA-A*03:01, HLA-A*11:01 and / or HLA-A*02:

01.

311. A polypeptide according to any one of claims 308-310, wherein the antigen binding domain has a binding affinity to the peptide conjugate / MHC complex that is greater than the binding affinity to the peptide or free targeted covalent inhibitor.

312. The polypeptide of any one of claims 307-311, wherein the antigen binding domain binds to the peptide conjugate / MHC complex with a dissociation constant (K D ) is 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.

313. The polypeptide of any one of claims 307-312, wherein the antigen binding domain binds to the peptide conjugate / MHC complex with a dissociation constant (K D ) is from about 0.01 nM to about 20 nM.

314. The polypeptide of any one of claims 308-313, wherein the antigen binding domain binds to the free targeted covalent inhibitor with a dissociation constant (K D ) is at least about 100 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 μM, at least about 10 μM, at least about 20 μM, at least about 30 μM, at least about 40 μM, at least about 50 μM, or at least 100 μM.

315. The polypeptide of any one of claims 308-313, wherein the dissociation constant (K) of the antigen binding domain binding to the free targeted covalent inhibitor is D ) is the K of the antibody or antigen-binding fragment binding to the peptide conjugate / MHC complex D at least about 10, 100, 10,000, or 100,000 times greater.

316. The polypeptide of any one of claims 308-315, wherein the antigen binding domain undetectably binds to the free targeted covalent inhibitor.

317. A polypeptide according to any one of claims 308-315, wherein the antigen binding domain has an IC of at least about 50 nM. 50 Binds to the free targeted covalent inhibitor.

318. The polypeptide of any one of claims 308-315, wherein the antigen binding domain binds to the free peptide conjugate with a dissociation constant (K D ) is greater than about 100 nM, greater than about 200 nM, greater than about 300 nM, greater than about 400 nM, greater than about 500 nM, greater than 1 μM, greater than 10 μM, greater than 20 μM, greater than 30 μM, greater than 40 μM, greater than 50 μM or greater than 100 μM.

319. The polypeptide of any one of claims 308-318, wherein the antigen binding domain binds to the free peptide conjugate with a dissociation constant (K D ) is the K of the antibody or antigen-binding fragment and the peptide conjugate / MHC complex D at least about 10, 100, 10,000, 100,000 times greater.

320. The polypeptide of any one of claims 308-318, wherein the antigen binding domain binds to the free peptide conjugate with a dissociation constant (K D ) is the K of the antibody or antigen-binding fragment and the peptide conjugate / MHC complex D At least 2.5 times of.

321. The polypeptide of claim 320, wherein the antigen binding domain binds to the free peptide conjugate with a dissociation constant (K D ) is greater than the K of the antibody or antigen-binding fragment and the peptide conjugate / MHC complex D , where MHC is HLA-A02:

01.

322. The polypeptide of claim 320, wherein the antigen binding domain binds to the free peptide conjugate with a dissociation constant (K D ) is the K of the antibody or antigen-binding fragment and the peptide conjugate / MHC complex D at least 2.5 times of that in which the MHC is HLA-A03:

01.

323. A polypeptide according to any one of claims 308-319, wherein the affinity of the antigen binding domain binding to 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, or at least 10,000 times the affinity of the antibody or antigen binding fragment to the free targeted covalent inhibitor or the free peptide conjugate.

324. The polypeptide of any one of claims 308-323, wherein the peptide conjugate is composed of a free targeted covalent inhibitor and a KRAS G12C Peptide, KRAS G12D Peptide, KRAS G12R Peptide or KRAS G12S Covalent reaction of peptides to form.

325. The polypeptide of claim 324, wherein the free targeted covalent inhibitor is osimertinib, ibrutinib, neratinib, sotolacib, ARS-853, ARS-1620, ARS-3248, MRTX849, JNJ74699157, LY3499446, LY3537982, MRTX-1257, JDQ443, MRTX-1133, RMC-6291, RMC-9805, GDC-6036, D-1553, 2E07, 6H05, SML-8-73-1 or BI 182391.

326. The polypeptide of claim 325, wherein the peptide conjugate is composed of sotolacib and KRAS G12C Covalent reaction of peptides to form.

327. The polypeptide of claim 324, wherein the peptide comprises or consists of an amino acid sequence of VVVGACGVGK, VVGACGVGK, or KLVVVGACGV.

328. A polypeptide according to any one of claims 166-327, wherein the antigen binding domain (i) is specific for a peptide conjugate / MHC complex comprising VVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*11:01 and / or a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*11:01, or (ii) is specific for a peptide conjugate / MHC complex comprising VVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:01 and / or a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:

01.

329. A polypeptide according to any one of claims 166-327, wherein the antigen binding domain is specific for a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:01 and / or a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*11:

01.

330. A polypeptide according to any one of claims 166-327, wherein the antigen binding domain is specific for a peptide conjugate / MHC complex comprising VVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*11:01, a peptide conjugate / MHC complex comprising VVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:01, a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*03:01, a peptide conjugate / MHC complex comprising VVVGACGVGK conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*11:01, and / or a peptide conjugate / MHC complex comprising KLVVVGACGV conjugated to a targeted covalent inhibitor or a fragment thereof presented by HLA-A*02:

01.

331. The polypeptide of any one of claims 166-330, wherein the polypeptide binds: (i) a peptide conjugate comprising a peptide consisting of the amino acid sequence VVGACGVGK / HLA-A*11:01 MHC complex and a peptide conjugate / HLA-A*11:01 MHC complex comprising a peptide consisting of the amino acid sequence VVVGACGVGK; (ii) a peptide conjugate comprising a peptide consisting of the amino acid sequence VVGACGVGK / HLA-A*03:01 MHC complex and a peptide conjugate / HLA-A*03:01 MHC complex comprising a peptide consisting of the amino acid sequence VVVGACGVGK; (iii) a peptide conjugate comprising a peptide consisting of the amino acid sequence VVGACGVGK / HLA-A*03:01 MHC complex and a peptide conjugate / HLA-A*11:01 MHC complex comprising a peptide consisting of the amino acid sequence VVGACGVGK; (iv) a peptide conjugate comprising a peptide consisting of the amino acid sequence VVVGACGVGK / HLA-A*03:01 MHC complex and A peptide conjugate comprising a peptide consisting of the amino acid sequence VVVGACGVGK / HLA-A*11:01 MHC complex; and / or (v) Peptide conjugate comprising a peptide consisting of the amino acid sequence KLVVVGACGV / HLA-A*02:01 MHC complex.

332. A polypeptide according to any one of claims 166-330, wherein the polypeptide binds to: a peptide conjugate comprising a peptide consisting of the amino acid sequence VVGACGVGK / HLA-A*03:01 MHC complex, and a peptide conjugate comprising a peptide consisting of the amino acid sequence VVVGACGVGK / HLA-A*03:01 MHC complex, and a peptide conjugate comprising a peptide consisting of the amino acid sequence VVGACGVGK / HLA-A*11:01 MHC complex, and a peptide conjugate comprising a peptide consisting of the amino acid sequence VVVGACGVGK / HLA-A*11:01 MHC complex, and a peptide conjugate comprising a peptide consisting of the amino acid sequence KLVVVGACGV / HLA-A*02:01 MHC complex.

333. The polypeptide of any one of claims 308-327, wherein the interface area between the polypeptide and the peptide conjugate / MHC complex is at least about or 334. The polypeptide of any one of claims 308-333, wherein the interface area of ​​the polypeptide with the MHC of the peptide conjugate / MHC complex is greater than the interface area of ​​the polypeptide with the peptide or the targeted covalent inhibitor.

335. The polypeptide of any one of claims 308-334, wherein the polypeptide forms a binding pocket at the interface between the VH and VL domains to accommodate the targeted covalent inhibitor of the peptide conjugate / MHC complex.

336. The polypeptide of any one of claims 308-335, wherein the polypeptide does not bind to the peptide conjugate / MHC complex via head-to-head coaxial interactions.

337. The polypeptide of any one of claims 308-336, wherein: (a) the polypeptide is bound to the peptide conjugate / MHC complex at an angle of about 10° to 60° between the axis of the MHC and the axis of the polypeptide; (b) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is about 10° to 60°; (c) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is different from the angle between the axis of the TCR and the axis of the peptide conjugate / MHC complex when the TCR is bound to the peptide conjugate / MHC complex; (d) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is about 10° to 60° or about 10° to 60° less than the angle between the axis of the TCR and the axis of the peptide conjugate / MHC complex when the TCR is bound to the peptide conjugate / MHC complex; (e) the polypeptide binds to the peptide conjugate / MHC complex at an angle that is different from the angle at which the T cell receptor binds to the peptide conjugate / MHC complex; and / or (f) the polypeptide binds to the peptide conjugate / MHC complex at an angle of about 10° to 60° or about 10° to 60° less than the angle at which a T cell receptor binds to the peptide conjugate / MHC complex.

338. A polypeptide according to any one of claims 308-337, wherein (a) the polypeptide is bound to the peptide conjugate / MHC complex at an angle of about 40° between the axis of the MHC and the axis of the polypeptide; (b) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is about 40°; (c) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is less than about 40° or approximately 40° compared to the angle between the axis of the TCR and the axis of the peptide conjugate / MHC complex when the TCR is bound to the peptide conjugate / MHC complex; and / or (d) the polypeptide binds to the peptide conjugate / MHC complex at an angle of about 40° or about 40° less than the angle at which a T cell receptor binds to the peptide conjugate / MHC complex.

339. The polypeptide of any one of claims 308-338, wherein the polypeptide contacts the α1 domain and the α2 domain of the heavy chain of the MHC of the peptide conjugate / MHC complex.

340. The polypeptide of any one of claims 308-339, wherein the polypeptide binds to an epitope of MHC, and wherein the epitope comprises one or more residues from a region comprising residues 62-66, 106-109 and / or 150-170 of MHC.

341. A polypeptide according to claim 340, wherein the polypeptide binds to an epitope of MHC, and wherein the epitope comprises one or more residues from a region comprising residues 62-66, 106-109 and / or 150-170 of HLA-A*03:01 or HLA-A*11:

01.

342. A polypeptide according to claim 340 or 341, wherein the polypeptide binds to an epitope of MHC, and wherein the epitope comprises 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 HLA-A*03:

01.

343. A polypeptide according to claim 342, wherein the VL domain of the antigen binding domain binds to an epitope of MHC, and wherein the epitope comprises 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 HLA-A*03:

01.

344. A polypeptide according to any one of claims 340-343, wherein the polypeptide binds to an epitope of MHC, and wherein the epitope comprises one or more residues selected from the group consisting of residues 62, 65, 66, 150, 151, 154, 155, 157 and 158 of HLA-A*03:

01.

345. A polypeptide according to claim 344, wherein the VH domain of the antigen binding domain binds to an epitope of MHC, and wherein the epitope comprises one or more residues selected from the group consisting of residues 62, 65, 66, 150, 151, 154, 155, 157 and 158 of HLA-A*03:

01.

346. A polypeptide according to claim 340 or 341, wherein the polypeptide binds to an epitope of MHC, and wherein the epitope comprises 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 HLA-A*11:

01.

347. A polypeptide according to claim 346, wherein the VL domain of the antigen binding domain binds to an epitope of MHC, and wherein the epitope comprises 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 HLA-A*11:

01.

348. A polypeptide according to claim 346 or 347, wherein the polypeptide binds to an epitope of MHC, and wherein the epitope comprises one or more residues selected from the group consisting of residues 62, 65, 66, 151, 154, 155 and 158 of HLA-A*11:

01.

349. A polypeptide according to claim 348, wherein the VH domain of the antigen binding domain binds to an epitope of MHC, and wherein the epitope comprises one or more residues selected from the group consisting of residues 62, 65, 66, 151, 154, 155 and 158 of HLA-A*11:

01.

350. The polypeptide of any one of claims 308-349, wherein the VH is connected to the VL via a linker.

351. The polypeptide of claim 350, wherein the linker comprises (G4S) n or (S4G) n , where n is any integer between 1 and 10.

352. A polypeptide according to claim 351, wherein the linker comprises a glycine-serine-alanine linker G4SA3 or a glycine-serine linker (G4S)4.

353. A polypeptide according to any one of claims 166-352, wherein the polypeptide is a complete antibody, a bispecific antibody, a multispecific antibody, an antigen binding (Fab) fragment, a Fab' fragment, a (Fab')2 fragment, Fd, Fv, dAb, a single domain fragment or a single monomer variable antibody domain, a dual affinity retargeting (DART) molecule, a diabody (Db), a single-chain diabody (scDb), a single-chain variable fragment (scFv), a bispecific T cell engaging antibody (BiTE), a bispecific killer cell engaging antibody (BiKE), a CrossMab, a camelid antibody, a trispecific binding partner, a chimeric antigen receptor (CAR), a monomer (also known as Adnectin), a DARPin, an anticalin, an affibody or an antibody-mimicking protein.

354. The polypeptide of claim 353, wherein the polypeptide is a bispecific antibody.

355. The polypeptide of claim 354, wherein the bispecific antibody is a bispecific T cell engaging antibody (BiTE).

356. A polypeptide according to any one of claims 166-355, wherein the polypeptide further comprises a second antigen binding domain that binds to a T cell surface marker.

357. A polypeptide according to claim 356, wherein the T cell surface marker is CD3ε, CD3γ, CD3δ, TCRα or TCRβ of CD3 small TCR.

358. The polypeptide of claim 356 or 357, wherein the antigen binding domain and the second antigen binding domain are connected by a linker.

359. The polypeptide of claim 358, wherein the linker comprises (G4S) n or (S4G) n , where n is any integer between 1 and 10.

360. The polypeptide of claims 358 and 359, wherein the linker is configured such that: (a) the polypeptide is bound to the peptide conjugate / MHC complex at an angle of about 10° to 60° between the axis of the MHC and the axis of the polypeptide; (b) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is about 10° to 60°; (c) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is different from the angle between the axis of the TCR and the axis of the peptide conjugate / MHC complex when the TCR is bound to the peptide conjugate / MHC complex; (d) when the polypeptide is bound to the peptide conjugate / MHC complex, the angle between the axis of the polypeptide and the axis of the peptide conjugate / MHC complex is about 10° to 60° or about 10° to 60° less than the angle between the axis of the TCR and the axis of the peptide conjugate / MHC complex when the TCR is bound to the peptide conjugate / MHC complex; (e) the polypeptide binds to the peptide conjugate / MHC complex at an angle that is different from the angle at which the T cell receptor binds to the peptide conjugate / MHC complex; and / or (f) the polypeptide binds to the peptide conjugate / MHC complex at an angle of about 10° to 60° or about 10° to 60° less than the angle at which a T cell receptor binds to the peptide conjugate / MHC complex.

361. The polypeptide of any one of claims 356-360, wherein the polypeptide comprises a first polypeptide chain comprising the antigen binding domain and a second polypeptide chain comprising the second antigen binding domain.

362. The polypeptide of claim 361, wherein the first polypeptide chain or the second polypeptide chain is further fused to a cytokine or a fragment thereof.

363. A polypeptide according to claim 362, wherein the cytokine comprises IL-2, IL-7, IL-15, IL-12, IL-18 or IL-21, or interferon (IFN).

364. A polypeptide according to any one of claims 307-363, wherein the peptide conjugate / MHC complex is presented on the surface of a cell.

365. The polypeptide of claim 364, wherein the cells 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 single cell.

366. The polypeptide of claim 364 or 365, wherein the peptide of the peptide conjugate / MHC complex is derived from an intracellular protein.

367. A polypeptide according to any one of claims 166-366, wherein the polypeptide is a binding partner according to any one of the preceding claims.

368. A polypeptide comprising an antigen binding domain that specifically binds to a peptide conjugate / MHC complex, wherein the peptide conjugate / MHC complex comprises: (c) a peptide conjugate comprising a peptide covalently linked to a targeted covalent inhibitor or a fragment thereof; and (d) MHC.

369. A pharmaceutical composition comprising a polypeptide according to any one of claims 166-368 and a pharmaceutically acceptable carrier.

370. A method of treating cancer in a subject who has been treated with a free targeted covalent inhibitor, the method comprising administering to the subject a polypeptide according to any one of claims 166-368 or a pharmaceutical composition according to claim 238.

371. The method of claim 370, wherein the subject is refractory to treatment with the free targeted covalent inhibitor.

372. A method of treating cancer in a subject, the method comprising administering to the subject a polypeptide according to any one of claims 166-368 or a pharmaceutical composition according to claim 369 after or simultaneously with administration of a small molecule drug.

373. A method of treating cancer in a subject, the method comprising administering to the subject a targeted covalent inhibitor, and The subject is administered the polypeptide of any one of claims 166-368 or the pharmaceutical composition of claim 369.

374. The method of claim 373, wherein the polypeptide of any one of claims 166-368 or the pharmaceutical composition of claim 369 is administered after or simultaneously with the targeted covalent inhibitor.

375. A method of identifying a T cell receptor (TCR) that recognizes a peptide conjugate / MHC complex according to any one of claims 167-374, the method comprising: (a) contacting a plurality of candidate TCRs with the peptide conjugate / MHC complex, and (b) identifying at least one TCR that binds to the peptide conjugate / MHC complex.

376. A method of identifying a T cell receptor (TCR) that recognizes a peptide conjugate / MHC complex according to any one of claims 167-374, the method comprising: (a) culturing T cells and antigen presenting cells (APCs) treated with a targeted covalent inhibitor, thereby generating peptide conjugate / MHC complex-specific T cells having a T cell receptor (TCR) that binds to the peptide conjugate / MHC complex; and (b) identifying at least one TCR from T cells specific for the peptide conjugate / MHC complex that binds to the peptide conjugate / MHC complex.

377. The method of claim 375 or 376, wherein the identification in (b) comprises selecting or isolating the at least one TCR.

378. The method of claim 375 or 377, wherein the plurality of candidate TCRs are a plurality of soluble TCRs or a plurality of TCRs expressed on the cell surface of a plurality of cells.

379. The method of claim 378, wherein the plurality of candidate TCRs are the plurality of TCRs expressed on the cell surface of the plurality of cells, and the identification in (b) comprises separating or selecting cells comprising at least one of the TCRs based on cell activation markers.

380. The method of claim 379, wherein the activation marker is a T cell activation marker.

381. The method of claim 380, wherein the T cell activation marker is CD26, CD27, CD28, CD30, CD154, CD40L, CD134, CD25, CD44, CD69, CD137 or KLRG1.

382. A T cell receptor (TCR) comprising at least one TCR identified in (b) according to any one of claims 375-381.

383. The TCR of claim 382, ​​wherein the TCR is a soluble TCR.

384. A TCR according to claim 382 or 383, wherein the TCR is a bispecific TCR.

385. A method of treating cancer, the method comprising prophylactically administering a peptide conjugate to a subject in need thereof before the subject develops cancer or before the subject is administered a drug, or both, wherein the peptide conjugate comprises a peptide covalently linked to a targeted covalent inhibitor or fragment thereof.

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