Antigen-binding proteins targeting shared neoantigens

Abstract

To provide antigen-binding proteins (ABPs) for use to discover and identify tumor-associated HLA-peptide complexes with highly positive predictive value, and for use in TCR-based immunotherapies targeting such complexes.SOLUTION: The disclosure provides an ABP that binds specifically to an HLA-peptide antigen comprising an HLA-restricted peptide forming a complex with an HLA Class I molecule, where the HLA-restricted peptide is located in the peptide binding groove of an α1 / α2 heterodimer portion of the HLA Class I molecule, where the HLA Class I molecule and the HLA-restricted peptide each have a specific amino acid sequence, and where the ABP comprises a T cell receptor (TCR) or antigen-binding fragment thereof.SELECTED DRAWING: Figure 12

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 62 / 936,303, filed November 15, 2019, and U.S. Provisional Patent Application No. 63 / 030,774, filed May 27, 2020, each of which is incorporated herein by reference in its entirety for all purposes.

[0002] Sequence Listing This application has been submitted electronically in ASCII format and contains a Sequence Listing, which is incorporated herein by reference in its entirety. The ASCII copy, created on November 13, 2020, is named GSO-080WO_SL.txt and is 6,925,625 bytes in size. [Background technology]

[0003] background It is recognized that MHC presents intracellularly processed protein fragments on the cell surface. In humans, MHC is called human leukocyte antigen or HLA. Specifically, MHC class I molecules are expressed on the surface of almost all nucleated cells in the body. MHC class I molecules are dimeric molecules containing a transmembrane heavy chain containing a peptide antigen-binding groove and a small extracellular chain called β2-microglobulin. MHC class I molecules present peptides derived from the degradation of cytosolic proteins by the proteasome, a multi-unit structure in the cytoplasm (Niedermann G., 2002. Curr Top Microbiol Immunol. 268:91-136; for bacterial antigen processing, see Wick MJ, and Ljunggren H G., 1999. Immunol Rev. 172:153-62). The cleaved peptides are transported into the lumen of the endoplasmic reticulum (ER) by a transporter associated with antigen processing (TAP), where they bind to the groove of the assembling class I molecule, and the resulting MHC / peptide complexes are transported to the plasma membrane to enable antigen presentation to T lymphocytes (Yewdell J W., 2001. Trends Cell Biol. 11:294-7; Yewdell J W. and Bennink J R., 2001. Curr Opin Immunol. 13:13-8). Alternatively, the cleaved peptides can be loaded onto MHC class I molecules in a TAP-independent manner, allowing for the presentation of extracellularly derived proteins through the process of cross-presentation.

[0004] MHC genes are highly polymorphic across species, containing multiple common alleles of individual genes. Therefore, a given MHC allele / peptide complex containing a particular HLA subtype and a particular peptide fragment presents a novel protein structure on the cell surface that can be targeted by novel antigen-binding proteins (e.g., TCRs or antigen-binding fragments). However, such TCR-based approaches first require the identification of the complex structure (peptide sequence and MHC subtype).

[0005] Tumor cells can express neoantigens and present them on their surface via MHC presentation. Such tumor-associated neoantigens, including novel protein structures formed by peptide-MHC subtype complexes, can be used to develop novel immunotherapy reagents for specific targeting of tumor cells. For example, tumor-associated antigens can be used to identify therapeutic antigen-binding proteins, such as TCRS, or antigen-binding fragments thereof. However, accurate identification of such neoantigens has been difficult.

[0006] Early methods have been proposed that incorporate mutation-based analysis using next-generation sequencing, RNA gene expression, and prediction of MHC binding affinity of candidate neoantigen peptides. 8 However, these proposed methods cannot model the entire epitope generation process, which involves many steps (e.g., TAP transport, proteasomal cleavage, and / or TCR recognition) in addition to gene expression and MHC binding. 9 As a result, existing methods can suffer from low positive predictive value (PPV).

[0007] Indeed, analyses of peptides presented by tumor cells conducted by several groups have shown that less than 5% of peptides predicted to be presented using gene expression and MHC binding affinity are actually found on tumor surface MHC. 10,11 This poor correlation between predicted binding and actual MHC presentation was further reinforced by the recent observation of a lack of improvement in the accuracy of predicting binding-restricted neoantigens in response to checkpoint inhibitors beyond the number of mutations alone. 12 .

[0008] This low positive predictive value (PPV) of existing methods for predicting presentation presents a problem for neoantigen-based immunotherapy design: if immunotherapies are designed using predictions with low PPV, many of them will be clinically ineffective.

[0009] Thus, there is a need for the discovery and identification of tumor-associated HLA-peptide complexes with high positive predictive value, and for the development of TCR-based immunotherapies that target such complexes. Summary of the Invention

[0010] overview Provided herein is an antigen binding protein (ABP) that specifically binds to an HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, wherein the HLA class I molecule and the HLA-restricted peptide are each selected from the HLA-peptide antigens set forth in any one of SEQ ID NOs: 10,755 to 29,364, and the ABP comprises a T cell receptor (TCR) or an antigen-binding fragment thereof.

[0011] In some embodiments, the HLA-restricted peptide is about 5 to 15 amino acids in length. In some embodiments, the HLA-restricted peptide is about 8 to 12 amino acids in length, optionally 8, 9, 10, 11, or 12 amino acids in length.

[0012] In some embodiments, the HLA-peptide antigen is selected from the group consisting of: HLA-A * RAS_G12D MHC class I antigen containing 11:01 and the restriction peptide VVVGADGVGK; HLA-A * RAS_G12V MHC class I antigen containing 11:01 and the restriction peptide VVVGAVGVGK; HLA-A * RAS_G12C MHC class I antigen containing 02:01 and the restriction peptide KLVVVGACGV; HLA-A * CTNNB1_S45P MHC class I antigen containing 03:01 and the restriction peptide TTAPPLSGK; HLA-A * RAS_G12D MHC class I antigen containing 11:01 and the restriction peptide VVGADGVGK; HLA-A *RAS_G12V MHC class I antigen containing 11:01 and the restriction peptide VVGAVGVGK; HLA-C * RAS_G12V MHC class I antigen containing 01:02 and the restriction peptide AVGVGKSAL; HLA-A * RAS_G12V MHC class I antigen containing 03:01 and the restriction peptide VVVGAVGVGK; HLA-A * TP53_K132N MHC class I antigen containing 24:02 and the restriction peptide TYSPALNNMF; HLA-A * CTNNB1_S37Y MHC class I antigen containing 02:01 and the restriction peptide YLDSGIHYGA; HLA-A * RAS_G12C MHC class I antigen containing 03:01 and the restriction peptide VVVGACGVGK; HLA-A*11: RAS_G12C MHC class I antigen containing 01 and the restriction peptide VVVGACGVGK; HLA-A * RAS_G12D MHC class I antigen containing 03:01 and the restriction peptide VVVGADGVGK; HLA-A * RAS_Q61H MHC class I antigen containing 01:01 and the constraint peptide ILDTAGHEEY; and A * TP53_R213L MHC class I antigen containing 02:01 and the restriction peptide YLDDRNTFL.

[0013] In some embodiments, the restriction peptide comprises a RAS_G12A mutation and wherein the HLA class I molecule is HLA-A*02:01; the restriction peptide comprises a RAS_G12A mutation and wherein the HLA class I molecule is HLA-A*02:06; the restriction peptide comprises a RAS_G12A mutation and wherein the HLA class I molecule is HLA-B*27:05; the restriction peptide comprises a RAS_G12A mutation and wherein the HLA class I molecule is HLA-B*35:01; the restriction peptide comprises The restriction peptide comprises a RAS_G12A mutation and wherein the HLA class I molecule is HLA-B*41:02; the restriction peptide comprises a RAS_G12A mutation and wherein the HLA class I molecule is HLA-B*48:01; the restriction peptide comprises a RAS_G12A mutation and wherein the HLA class I molecule is HLA-C*08:03; the restriction peptide comprises a RAS_G12C mutation and wherein the HLA class I molecule is HLA-A*02:01; the restriction peptide comprises a RAS_G12C mutation and wherein the HLA class I molecule is HLA- wherein the HLA class I molecule is HLA-A*02:01; wherein the restriction peptide comprises a RAS_G12C mutation and wherein the HLA class I molecule is HLA-A*03:02; wherein the restriction peptide comprises a RAS_G12C mutation and wherein the HLA class I molecule is HLA-A*68:01; wherein the restriction peptide comprises a RAS_G12C mutation and wherein the HLA class I molecule is HLA-B*27:05; wherein the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is whether the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*02:05; whether the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*03:01; whether the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*11:01; whether the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*11:01;The restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*26:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*31:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*68:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*07:02; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*07:02; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*08:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*13:02; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*15:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*27:05; the restriction peptide comprises a RAS_G12D mutation and wherein The HLA class I molecule is HLA-B*35:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*37:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*38:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*40:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*40:01. whether the restriction peptide comprises a RAS_G12D mutation and the HLA class I molecule is HLA-B*44:02; whether the restriction peptide comprises a RAS_G12D mutation and the HLA class I molecule is HLA-B*44:03; whether the restriction peptide comprises a RAS_G12D mutation and the HLA class I molecule is HLA-B*48:01; whether the restriction peptide comprises a RAS_G12D mutation and the HLA class I molecule is HLA-B*50:01;The restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*57:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*01:02; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*02:02; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*03:03; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*04:04; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*03:04; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*04:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*05:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*07:04; the restriction peptide comprises a RAS_G12D mutation and wherein The HLA class I molecule is HLA-C*08:02; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*08:03; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*16:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*17:01; the restriction peptide comprises a RAS_G12R mutation and wherein the HLA class I molecule is HLA-C*18:02; whether the restriction peptide comprises a RAS_G12R mutation and the HLA class I molecule is HLA-C*07:04; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-A*02:01; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-A*02:05; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-A*02:06;The restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*03:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*03:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*11:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*11:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*11:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*11:01 the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*25:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*26:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*30:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*31:01; the restriction peptide comprises a RAS_G12V mutation and wherein The HLA class I molecule is HLA-A*31:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*32:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*68:02; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*07:02; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-B*13:02; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-B*14:02; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-B*15:01; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-B*27:05;The restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*39:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*40:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*40:02; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*41:02; the restriction peptide comprises a RAS_G the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*44:05; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*50:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*51:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-C*01:02; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-C*01:02. the HLA class I molecule is HLA-C*01:02; the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-C*03:03; the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-C*03:04; the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-C*08:02; the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA -C*14:02; or the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-C*17:01; or the restriction peptide comprises a KRAS_G13D mutation and the HLA class I molecule is HLA-A*02:01; or the restriction peptide comprises a KRAS_G13D mutation and the HLA class I molecule is HLA-B*07:02; or the restriction peptide comprises a KRAS_G13D mutation and the HLA class I molecule is HLA-B*08:01;the restriction peptide comprises a KRAS_G13D mutation and wherein the HLA class I molecule is HLA-B*35:01; the restriction peptide comprises a KRAS_G13D mutation and wherein the HLA class I molecule is HLA-B*35:03; the restriction peptide comprises a KRAS_G13D mutation and wherein the HLA class I molecule is HLA-B*35:08; the restriction; The bundle peptide contains a KRAS_G13D mutation and the HLA class I molecule is HLA-B*38:01; the restriction peptide contains a KRAS_G13D mutation and the HLA class I molecule is HLA-C*04:01; the restriction peptide contains a KRAS_Q61H mutation and the HLA class I molecule is HLA-A*01:01; the restriction peptide contains a KRAS_Q61H mutation and the HLA class I molecule is HLA-A*02:01; the restriction peptide contains a KRAS_Q6 the restriction peptide comprises a KRAS_Q61H mutation and wherein the HLA class I molecule is HLA-A*23:01; the restriction peptide comprises a KRAS_Q61H mutation and wherein the HLA class I molecule is HLA-A*29:01; the restriction peptide comprises a KRAS_Q61H mutation and wherein the HLA class I molecule is HLA-A*30:02; the restriction peptide comprises a KRAS_Q61H mutation and wherein the HLA class I molecule is HLA-A*33:01; the restriction peptide comprises a KRAS_Q61H mutation and wherein the HLA class I molecule is HLA-A*30:02. the HLA class I molecule is HLA-A*68:01; the restriction peptide comprises a KRAS_Q61H mutation and wherein the HLA class I molecule is HLA-B*07:02; the restriction peptide comprises a KRAS_Q61H mutation and wherein the HLA class I molecule is HLA-B*08:01; the restriction peptide comprises a KRAS_Q61H mutation and wherein the HLA class I molecule is HLA-B*18:01; the restriction peptide comprises a KRAS_Q61H mutation and wherein the HLA class I molecule is HLA A-B*35:01; whether the restriction peptide comprises a KRAS_Q61H mutation and wherein the HLA class I molecule is HLA-B*38:01; whether the restriction peptide comprises a KRAS_Q61H mutation and wherein the HLA class I molecule is HLA-B*40:01; whether the restriction peptide comprises a KRAS_Q61H mutation and wherein the HLA class I molecule is HLA-B*44:02; whether the restriction peptide comprises a KRAS_Q61H mutation and wherein the HLA class I molecule is HLA-C*03:04;The restricted peptide contains a KRAS_Q61H mutation and the HLA class I molecule is HLA-C*05:01; or the restricted peptide contains a KRAS_Q61H mutation and the HLA class I molecule is HLA-C*08:02.

[0014] In some embodiments, the restriction peptide comprises a KRAS_G13D mutation and wherein the HLA class I molecule is C * 08:02 or A * 11:01; the restricted peptide contains the KRAS_Q61K mutation, and wherein the HLA class I molecule is A * 01:01; the restricted peptide contains the NRAS_Q61K mutation, and wherein the HLA class I molecule is A *the restriction peptide comprises a TP53_R249M mutation and the HLA class I molecule is B*35:12, B*35:03, or B*35:01; the restriction peptide comprises a CTNNB1_S45P mutation and the HLA class I molecule is A*03:01, A*11:01, A*68:01, or A*03:02; the restriction peptide comprises a CTNNB1_S45F mutation and the HLA class I molecule is A*03:01, A*11:01, or A*68:01; The restriction peptide comprises an ERBB2_Y772_A775dup mutation and the HLA class I molecule is B*18:01; the restriction peptide comprises a KRAS_G12D mutation and the HLA class I molecule is A*11:01, A*03:01, or C*08:02; the restriction peptide comprises an NRAS_G12D mutation and the HLA class I molecule is A*11:01, A*03:01, or C*08:02; the restriction peptide comprises a KRAS_Q61R mutation and the HLA class I molecule is the A class I molecule is A*01:01; the restriction peptide comprises an NRAS_Q61R mutation and wherein the HLA class I molecule is A*01:01; the restriction peptide comprises a CTNNB1_T41A mutation and wherein the HLA class I molecule is A*03:01, A*03:02, A*11:01, B*15:10, C*03:03, or C*03:04; the restriction peptide comprises a TP53_K132N mutation and wherein the HLA class I molecule is A*24:02 or A*23:01; the restriction the peptide comprises a KRAS_G12A mutation and the HLA class I molecule is A*03:01 or A*11:01; the restriction peptide comprises a KRAS_Q61L mutation and the HLA class I molecule is A*01:01; the restriction peptide comprises an NRAS_Q61L mutation and the HLA class I molecule is A*01:01; the restriction peptide comprises a TP53_R213L mutation and the HLA class I molecule is A*02:07, C*08:02, or A*02:01;The restriction peptide comprises a BRAF_G466V mutation and the HLA class I molecule is B*15:01 or B*15:03; the restriction peptide comprises a KRAS_G12V mutation and the HLA class I molecule is A*03:01, A*03:02, A*11:01, or C*01:02; the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is A*01:01; the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is A*01:01; the restriction peptide comprises a NRAS_Q61H mutation and the HLA class I molecule is A*01:01; the restriction peptide comprises a CTNNB1_S37F mutation and the HLA class I molecule is A*01:01, A*23:01, A*24:02, B*15:10, B*39:06, C*05:01, C*14:02, or C*14:03; the restriction peptide comprises a TP53_S127Y mutation and the HLA class I molecule is A*11 :01 or A*03:01; the restriction peptide comprises a TP53_K132E mutation and the HLA class I molecule is A*24:02, C*14:03, or A*23:01; the restriction peptide comprises a KRAS_G12C mutation and the HLA class I molecule is A*02:01, A*11:01, or A*03:01; the restriction peptide comprises an NRAS_G12C mutation and the HLA class I molecule is the restriction peptide comprises an EGFR_L858R mutation and the HLA class I molecule is A*11:01 or A*03:01; the restriction peptide comprises a TP53_Y220C mutation and the HLA class I molecule is A*02:01; or the restriction peptide comprises a TP53_R175H mutation and the HLA class I molecule is A*02:01;

[0015] In some embodiments, the HLA-peptide antigen is selected from the following: CTNNB1_S45P MHC class I antigen comprising A*11:01 and the restriction peptide TTAPPLSGK; CTNNB1_T41AMHC class I antigen comprising A*11:01 and the restriction peptide ATAPSLSGK; RAS_G12D MHC class I antigen comprising A*11:01 and the restriction peptide VVVGADGVGK; RAS_G12V MHC class I antigen comprising A*03:01 and the restriction peptide VVGAVGVGK; RAS_G12V MHC class I antigen comprising A*03:01 and the restriction peptide VVVGAVGVGK; RAS_G12V MHC class I antigen comprising A*11:01 and the restriction peptide VVGAVGVGK; RAS_G12V MHC class I antigen comprising A*11:01 and the restriction peptide VVVGAVGVGK; MHC class I antigen; KRAS_Q61R MHC class I antigen comprising A*01:01 and the restriction peptide ILDTAGREEY; and TP53_R213L MHC class I antigen comprising A*02:01 and the restriction peptide YLDDRNTFL.

[0016] In some embodiments, the HLA-restricted peptide comprises a RAS G12 mutation. In some embodiments, the G12 mutation is a G12C, G12D, G12V, or G12A mutation. In some embodiments, the HLA-peptide antigen comprises an HLA class I molecule selected from HLA-A*02:01, HLA-A*11:01, HLA-A*31:01, HLA-C*01:02, and HLA-A*03:01. In some embodiments, the RAS G12 mutation is any one or more of the following: a KRAS mutation, an NRAS mutation, and an HRAS mutation. In some embodiments, the HLA-peptide antigen is selected from the following: a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV; a RAS_G12C MHC class I antigen comprising HLA-A*03:01 and the restriction peptide VVVGACGVGK; a RAS_G12C MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGACGVGK; a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK; a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVGADGVGK; a RAS_G12D MHC class I antigen comprising HLA-A*03:01 and the restriction peptide VVVGADGVGK. MHC class I antigens; RAS_G12V MHC class I antigens comprising HLA-A*11:01 and the restriction peptide VVVGAVGVGK; RAS_G12V MHC class I antigens comprising HLA-A*31:01 and the restriction peptide VVVGAVGVGK; RAS_G12V MHC class I antigens comprising HLA-A*11:01 and the restriction peptide VVGAVGVGK; RAS_G12V MHC class I antigens comprising HLA-C*01:02 and the restriction peptide AVGVGKSAL; and RAS_G12V MHC class I antigens comprising HLA-A*03:01 and the restriction peptide VVVGAVGVGK.In some embodiments, the HLA-peptide antigen is selected from the following: RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV; RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK; RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVGADGVGK; RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGAVGVGK; RAS_G12V MHC class I antigen comprising HLA-A*31:01 and the restriction peptide VVVGAVGVGK; RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVGAVGVGK. MHC class I antigens; RAS_G12V MHC class I antigens comprising HLA-C*01:02 and the restriction peptide AVGVGKSAL; and RAS_G12V MHC class I antigens comprising HLA-A*03:01 and the restriction peptide VVVGAVGVGK. In some embodiments, the HLA-peptide antigens are selected from: RAS_G12C MHC class I antigens comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV; RAS_G12D MHC class I antigens comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK; and RAS_G12V MHC class I antigens comprising HLA-A*11:01 and the restriction peptide VVVGAVGVGK. In some embodiments, the HLA-peptide antigen is RAS_G12C MHC class I antigens comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV. In some embodiments, the HLA-peptide antigen is a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK. In some embodiments, the HLA-peptide antigen is a RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGAVGVGK.

[0017] In some embodiments, the HLA-restricted peptide comprises a RAS Q61 mutation. In some embodiments, the Q61 mutation is a Q61H, Q61K, Q61R, or Q61L mutation. In some embodiments, the HLA-peptide antigen is a RAS_Q61H MHC class I antigen comprising HLA-A*01:01 and the restricted peptide ILDTAGHEEY.

[0018] In some embodiments, the HLA-restricted peptide comprises a TP53 mutation. In some embodiments, the TP53 mutation comprises a R213L, S127Y, Y220C, R175H, or R249M mutation. In some embodiments, the HLA-peptide antigen is a TP53 R213L MHC class I antigen comprising A*02:01 and the restriction peptide YLDDRNTFL.

[0019] In some embodiments, the antigen binding protein binds to an HLA-peptide antigen through at least one contact point with an HLA class I molecule and through at least one contact point with an HLA-restricted peptide.

[0020] In some embodiments, the antigen binding protein binds to a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV, and the ABP binds to the RAS_G12C MHC class I antigen with higher affinity than an HLA-peptide antigen comprising a different RAS G12 mutation. In some embodiments, the ABP binds to a RAS_G12C MHC class I antigen with higher affinity than an HLA-peptide antigen comprising the restriction peptide KLVVVGAVGV and an HLA-A2 molecule. In some embodiments, the ABP does not bind to an HLA-peptide antigen comprising the restriction peptide KLVVVGAVGV and an HLA-A2 molecule.

[0021] In some embodiments, the antigen binding protein is linked to a scaffold, optionally the scaffold comprises serum albumin or Fc, optionally the Fc is human and is an Fc of the IgG (IgG1, IgG2, IgG3, IgG4), IgA (IgA1, IgA2), IgD, IgE, or IgM isotype.

[0022] In some embodiments, the antigen binding protein is linked to the scaffold via a linker, optionally the linker is a peptide linker, optionally the peptide linker is the hinge region of a human antibody.

[0023] In some embodiments, the TCR or antigen-binding portion thereof comprises a TCR variable region. In some aspects, the TCR or antigen-binding portion thereof comprises one or more TCR complementarity-determining regions (CDRs). In some aspects, the TCR comprises an alpha chain and a beta chain. In some aspects, the TCR comprises a gamma chain and a delta chain. In some aspects, the TCR comprises a single-chain TCR (scTCR). In some aspects, the TCR comprises a recombinant TCR sequence. In some aspects, the TCR comprises a human TCR sequence, optionally a fully human TCR sequence. In some aspects, the TCR comprises a modified TCR alpha constant (TRAC) region, a modified TCR beta constant (TRBC) region, or a modified TRAC region and a modified TRBC region.

[0024] In some embodiments, the antigen binding protein comprises a modification that increases its half-life.

[0025] In some embodiments, the antigen binding protein is part of a chimeric antigen receptor (CAR) comprising an extracellular portion comprising the antigen binding protein and an intracellular signaling domain. In some embodiments, the intracellular signaling domain comprises an ITAM. In some embodiments, the intracellular signaling domain comprises the signaling domain of the zeta chain of the CD3 zeta (CD3) chain.

[0026] In some embodiments, the antigen binding protein further comprises a transmembrane domain connecting the extracellular domain and the intracellular signaling domain, hi some embodiments, the transmembrane domain comprises the transmembrane portion of CD28.

[0027] In some embodiments, the antigen binding protein further comprises an intracellular signaling domain of a T cell costimulatory molecule, hi some embodiments, the T cell costimulatory molecule is CD28, 4-1BB, OX-40, ICOS, or any combination thereof.

[0028] Also provided herein is a medicament comprising any one of the ABPs described herein.

[0029] Also provided herein is an ABP for use in treating cancer, optionally wherein the cancer expresses or is predicted to express an HLA-peptide antigen, a medicament comprising any one of the ABPs described herein. In some embodiments, the cancer is selected from a solid tumor and a hematological tumor.

[0030] Also provided herein are antigen binding proteins (ABPs) that compete for binding with any one of the ABPs described herein.

[0031] Also provided herein are antigen binding proteins (ABPs) that bind to the same HLA-peptide antigen epitope as bound by any one of the ABPs described herein.

[0032] Also provided herein are engineered cells expressing a receptor comprising an antigen-binding protein of any one of the ABPs described herein. In some embodiments, the engineered cells are T cells. In some embodiments, the T cells are selected from the group consisting of naive T (TN) cells, effector T cells (TEFF), memory T cells, stem cell memory T cells (TSCM), central memory T cells (TCM), effector memory T cells (TEM), terminally differentiated effector memory T cells, tumor-infiltrating lymphocytes (TIL), immature T cells, mature T cells, helper T cells, cytotoxic T cells, mucosal-associated invariant T (MALT) cells, regulatory T cells (Treg), TH1 cells, TH2 cells, TH3 cells, TH17 cells, TH9 cells, TH22 cells, follicular helper T cells, natural killer T cells (NKT), alpha-beta T cells, and gamma-delta T cells. In some embodiments, the T cells are cytotoxic T cells (CTL). In some embodiments, the engineered cells are human cells or human-derived cells. In some embodiments, the engineered cells are autologous cells of the subject. In some embodiments, the subject is known or suspected to have cancer. In some embodiments, the autologous cells are cells isolated from the subject. In some embodiments, the isolated cells are ex vivo cultured cells, and optionally the in vivo cultured cells are stimulated cells. In some embodiments, the autologous cells are in vivo engineered cells. In some embodiments, the antigen binding protein is expressed by a heterologous promoter. In some embodiments, the ABP comprises a T cell receptor (TCR) or an antigen-binding portion thereof, wherein a polynucleotide encoding the T cell receptor (TCR) or an antigen-binding portion thereof is inserted into an endogenous TCR locus. In some embodiments, the engineered cells do not express an endogenous ABP.

[0033] Also provided herein is an isolated polynucleotide encoding any one of the ABPs described herein, or a set of polynucleotides encoding any one of the ABPs described herein. Also provided herein is a vector or a set of vectors comprising any one of the polynucleotides or sets of polynucleotides described herein. Also provided herein is a virus comprising any one of the polynucleotides or sets of polynucleotides described herein. In some embodiments, the virus is a filamentous phage.

[0034] Also provided herein is a yeast cell comprising any one of the polynucleotides or set of polynucleotides described herein.

[0035] Also provided herein is a host cell comprising any one of the polynucleotides or set of polynucleotides described herein, optionally wherein the host cell is a CHO or HEK293, and optionally wherein the host cell is a T cell.

[0036] Also provided herein is a method of producing an antigen binding protein, comprising expressing the antigen binding protein using any one of the host cells described herein and isolating the expressed antigen binding protein.

[0037] Also provided herein are pharmaceutical compositions comprising any one of the antigen binding proteins described herein and a pharmaceutically acceptable excipient.

[0038] Also provided herein is a method of treating cancer in a subject comprising administering to the subject any one of the antigen binding proteins described herein, any one of the engineered cells described herein, or any one of the pharmaceutical compositions described herein, optionally wherein the cancer is selected from a solid tumor and a hematological tumor.

[0039] Also provided herein is a method of stimulating an immune response in a subject comprising administering to the subject any one of the antigen binding proteins described herein, any one of the engineered cells described herein, or any one of the pharmaceutical compositions described herein, optionally wherein the cancer is selected from a solid tumor and a hematological tumor.

[0040] Also provided herein is a method of killing target cells in a subject comprising administering to the subject any one of the antigen binding proteins described herein, any one of the engineered cells described herein, or any one of the pharmaceutical compositions described herein, optionally wherein the cancer is selected from a solid tumor and a hematological tumor.

[0041] In some embodiments, the subject is a human subject.

[0042] In some embodiments, the cancer expresses or is predicted to express an HLA-peptide antigen or HLA class I molecule described in any one of the embodiments. In some embodiments, the cancer expresses or is predicted to express an HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, the HLA-restricted peptide being located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, wherein the HLA class I molecule and the HLA-restricted peptide are each selected from HLA-peptide antigens such as those set forth in any one of SEQ ID NOS: 10,755-29,364, and wherein an ABP binds to the HLA-peptide antigen.In some embodiments, the HLA-peptide antigen is selected from the group consisting of: a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV; a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK; a RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGAVGVGK; a CTNNB1_S45P MHC class I antigen comprising HLA-A*03:01 and the restriction peptide TTAPPLSGK; a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVGADGVGK; a RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVGAVGVGK. MHC class I antigens; RAS_G12V MHC class I antigens containing HLA-C*01:02 and the restriction peptide AVGVGKSAL; RAS_G12V MHC class I antigens containing HLA-A*03:01 and the restriction peptide VVVGAVGVGK; TP53_K132N MHC class I antigens containing HLA-A*24:02 and the restriction peptide TYSPALNNMF; CTNNB1_S37Y MHC class I antigens containing HLA-A*02:01 and the restriction peptide YLDSGIHYGA; RAS_G12C MHC class I antigens containing HLA-A*03:01 and the restriction peptide VVVGACGVGK; RAS_G12C MHC class I antigens containing HLA-A*11:01 and the restriction peptide VVVGACGVGK; RAS_G12D MHC class I antigens containing HLA-A*03:01 and the restriction peptide VVVGADGVGK MHC class I antigens; RAS_Q61H MHC class I antigens comprising HLA-A*01:01 and the restriction peptide ILDTAGHEEY; and TP53_R213L MHC class I antigens comprising A*02:01 and the restriction peptide YLDDRNTFL.

[0043] In some embodiments, the restriction peptide comprises a RAS_G12A mutation and wherein the HLA class I molecule is HLA-A*02:01; the restriction peptide comprises a RAS_G12A mutation and wherein the HLA class I molecule is HLA-A*02:06; the restriction peptide comprises a RAS_G12A mutation and wherein the HLA class I molecule is HLA-B*27:05; the restriction peptide comprises a RAS_G12A mutation and wherein the HLA class I molecule is HLA-B*35:01; the restriction peptide comprises a RAS_G12A mutation and wherein the HLA class I molecule is HLA-B*35:01. the restriction peptide comprises a RAS_G12A mutation and the HLA class I molecule is HLA-B*41:02; the restriction peptide comprises a RAS_G12A mutation and the HLA class I molecule is HLA-B*48:01; the restriction peptide comprises a RAS_G12A mutation and the HLA class I molecule is HLA-C*08:03; the restriction peptide comprises a RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; the restriction peptide comprises a RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; and wherein the HLA class I molecule is HLA-A*02:01; wherein the restriction peptide comprises a RAS_G12C mutation and wherein the HLA class I molecule is HLA-A*03:01; wherein the restriction peptide comprises a RAS_G12C mutation and wherein the HLA class I molecule is HLA-A*03:02; wherein the restriction peptide comprises a RAS_G12C mutation and wherein the HLA class I molecule is HLA-A*68:01; wherein the restriction peptide comprises a RAS_G12C mutation and wherein the HLA class I molecule is , HLA-B*27:05; whether the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*02:01; whether the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*02:05; whether the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*03:01; whether the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*11:01;The restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*11:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*26:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*31:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*68:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-A*68:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*07:02; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*08:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*13:02; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*15:01; the restriction peptide comprises a RAS_G12D mutation and wherein The HLA class I molecule is HLA-B*27:05; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*35:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*37:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*38:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*38:01. whether the restriction peptide comprises a RAS_G12D mutation and the HLA class I molecule is HLA-B*40:01; whether the restriction peptide comprises a RAS_G12D mutation and the HLA class I molecule is HLA-B*40:02; whether the restriction peptide comprises a RAS_G12D mutation and the HLA class I molecule is HLA-B*44:02; whether the restriction peptide comprises a RAS_G12D mutation and the HLA class I molecule is HLA-B*44:03; whether the restriction peptide comprises a RAS_G12D mutation and the HLA class I molecule is HLA-B*48:01;The restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*50:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-B*57:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*01:02; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*02:02; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*03:02; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*03:03; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*03:04; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*04:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*05:01; the restriction peptide comprises a RAS_G12D mutation and wherein The HLA class I molecule is HLA-C*07:04; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*08:02; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*08:03; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*16:01; the restriction peptide comprises a RAS_G12D mutation and wherein the HLA class I molecule is HLA-C*16:01. whether the restriction peptide comprises a RAS_G12R mutation and the HLA class I molecule is HLA-B*41:02; whether the restriction peptide comprises a RAS_G12R mutation and the HLA class I molecule is HLA-C*07:04; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-A*02:01; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-A*02:05;The restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*02:06; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*03:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*03:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*11:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*11:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*11:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*25:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*26:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*30:01; the restriction peptide comprises a RAS_G12V mutation and wherein The HLA class I molecule is HLA-A*31:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*31:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*32:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-A*68:02; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-B*07:02; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-B*08:01; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-B*13:02; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-B*14:02; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-B*15:01;The restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*27:05; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*39:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*40:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*40:02; the restriction peptide comprises a RAS_G the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*41:02; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*44:05; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*50:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*51:01; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-B*50:01. the HLA class I molecule is HLA-C*01:02; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-C*01:02; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-C*03:03; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA-C*03:04; the restriction peptide comprises a RAS_G12V mutation and wherein the HLA class I molecule is HLA -C*08:02; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-C*14:02; whether the restriction peptide comprises a RAS_G12V mutation and the HLA class I molecule is HLA-C*17:01; whether the restriction peptide comprises a KRAS_G13D mutation and the HLA class I molecule is HLA-A*02:01; whether the restriction peptide comprises a KRAS_G13D mutation and the HLA class I molecule is HLA-B*07:02;the restriction peptide comprises a KRAS_G13D mutation and wherein the HLA class I molecule is HLA-B*08:01; the restriction peptide comprises a KRAS_G13D mutation and wherein the HLA class I molecule is HLA-B*35:01; the restriction peptide comprises a KRAS_G13D mutation and wherein the HLA class I molecule is HLA-B*35:03; whether the restriction peptide comprises a KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:08; whether the restriction peptide comprises a KRAS_G13D mutation and the HLA class I molecule is HLA-B*38:01; whether the restriction peptide comprises a KRAS_G13D mutation and the HLA class I molecule is HLA-C*04:01; whether the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-A*01:01; whether the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-A*02:01; whether the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-A*23:01; whether the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-A*29:01; whether the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-A*30:02; whether the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-A*33:01; whether the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-A*68:01; whether the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-B*07:02; whether the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-B*08:01; whether the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-B*18:01; whether the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-B*35:01; whether the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-B*38:01; whether the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-B*40:01;The restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-B*44:02; the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-C*03:04; the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-C*05:01; or the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is HLA-C*08:02.

[0044] In some embodiments, the restriction peptide comprises a KRAS_G13D mutation and the HLA class I molecule is C*08:02 or A*11:01; the restriction peptide comprises a KRAS_Q61K mutation and the HLA class I molecule is A*01:01; the restriction peptide comprises an NRAS_Q61K mutation and the HLA class I molecule is A*01:01; the restriction peptide comprises a TP53_R249M mutation and the HLA class I molecule is B*35:12, B*35:03 or B*35:01; the restriction peptide comprises a CTNNB1_S45P mutation and the HLA class I molecule is A*03:01, A*11:01, A*68:01, or A*03:02; the restriction peptide comprises a CTNNB1_S45F mutation and the HLA class I molecule is A*03:01, A*11:01, or A*68:01; the restriction peptide comprises an ERBB2_Y772_A775dup mutation and the HLA class I molecule is B*18:01; The restriction peptide comprises a KRAS_G12D mutation and wherein the HLA class I molecule is A*11:01, A*03:01, or C*08:02; the restriction peptide comprises an NRAS_G12D mutation and wherein the HLA class I molecule is A*11:01, A*03:01, or C*08:02; the restriction peptide comprises a KRAS_Q61R mutation and wherein the HLA class I molecule is A*01:01; the restriction peptide comprises an NRAS_Q61R mutation and wherein the HLA class I molecule is , A*01:01; the restriction peptide comprises a CTNNB1_T41A mutation and wherein the HLA class I molecule is A*03:01, A*0302, A*11:01, B*15:10, C*03:03, or C*03:04; the restriction peptide comprises a TP53_K132N mutation and wherein the HLA class I molecule is A*24:02 or A*23:01; the restriction peptide comprises a KRAS_G12A mutation and wherein the HLA class I molecule is A*03:01 or A*11:01;The restriction peptide comprises a KRAS_Q61L mutation and the HLA class I molecule is A*01:01; the restriction peptide comprises an NRAS_Q61L mutation and the HLA class I molecule is A*01:01; the restriction peptide comprises a TP53_R213L mutation and the HLA class I molecule is A*02:07, C*08:02, or A*02:01; the restriction peptide comprises a BRAF_G466V mutation and the HLA class I molecule is B*15:01, or B*1 the restriction peptide comprises a KRAS_G12V mutation and the HLA class I molecule is A*03:01, A*03:02, A*11:01, or C*01:02; the restriction peptide comprises a KRAS_Q61H mutation and the HLA class I molecule is A*01:01; the restriction peptide comprises an NRAS_Q61H mutation and the HLA class I molecule is A*01:01; the restriction peptide comprises a CTNNB1_S37F mutation and the HLA class I molecule is A*01:01. is A*01:01, A*23:01, A*24:02, B*15:10, B*39:06, C*05:01, C*14:02, or C*14:03; the restriction peptide comprises a TP53_S127Y mutation and the HLA class I molecule is A*11:01 or A*03:01; the restriction peptide comprises a TP53_K132E mutation and the HLA class I molecule is A*24:02, C*14:03, or A*23:01; the restriction peptide comprises a KRAS_G12C mutation and the HLA class I molecule is A*11:01 or A*03:01. wherein the HLA class I molecule is A*02:01, A*11:01, or A*03:01; wherein the restriction peptide comprises an NRAS_G12C mutation and wherein the HLA class I molecule is A*02:01, A*11:01, or A*03:01; wherein the restriction peptide comprises an EGFR_L858R mutation and wherein the HLA class I molecule is A*11:01 or A*03:01; wherein the restriction peptide comprises a TP53_Y220C mutation and wherein the HLA class I molecule is A*02:01;Alternatively, the restriction peptide comprises the TP53_R175H mutation, and the HLA class I molecule is A*02:01;

[0045] In some embodiments, the HLA-peptide antigen is selected from: CTNNB1_S45P MHC class I antigen comprising A*11:01 and the restriction peptide TTAPPLSGK; CTNNB1_T41A MHC class I antigen comprising A*11:01 and the restriction peptide ATAPSLSGK; RAS_G12D MHC class I antigen comprising A*11:01 and the restriction peptide VVVGADGVGK; RAS_G12V MHC class I antigen comprising A*03:01 and the restriction peptide VVGAVGVGK; RAS_G12V MHC class I antigen comprising A*03:01 and the restriction peptide VVVGAVGVGK; RAS_G12V MHC class I antigen comprising A*11:01 and the restriction peptide VVGAVGVGK; RAS_G12V MHC class I antigen comprising A*11:01 and the restriction peptide VVVGAVGVGK; RAS_G12V MHC class I antigen; KRAS_Q61R MHC class I antigen comprising A*01:01 and the restriction peptide ILDTAGREEY; and TP53_R213L MHC class I antigen comprising A*02:01 and the restriction peptide YLDDRNTFL.

[0046] In some embodiments, the HLA-peptide antigen comprises an HLA-restricted peptide, which is a peptide fragment of RAS comprising a RAS G12 mutation. In some embodiments, the G12 mutation is a G12C, G12D, G12V, or G12A mutation. In some embodiments, the HLA-peptide antigen comprises an HLA class I molecule selected from HLA-A*02:01, HLA-A*11:01, HLA-A*31:01, HLA-C*01:02, and HLA-A*03:01. In some embodiments, the RAS G12 mutation is any one or more of the following: a KRAS mutation, an NRAS mutation, and an HRAS mutation. In some embodiments, the HLA-peptide antigen is selected from the following: a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV; a RAS_G12C MHC class I antigen comprising HLA-A*03:01 and the restriction peptide VVVGACGVGK; a RAS_G12C MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGACGVGK; a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK; a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVGADGVGK; a RAS_G12D MHC class I antigen comprising HLA-A*03:01 and the restriction peptide VVVGADGVGK. MHC class I antigens; RAS_G12V MHC class I antigens comprising HLA-A*11:01 and the restriction peptide VVVGAVGVGK; RAS_G12V MHC class I antigens comprising HLA-A*31:01 and the restriction peptide VVVGAVGVGK; RAS_G12V MHC class I antigens comprising HLA-A*11:01 and the restriction peptide VVGAVGVGK; RAS_G12V MHC class I antigens comprising HLA-C*01:02 and the restriction peptide AVGVGKSAL; and RAS_G12V MHC class I antigens comprising HLA-A*03:01 and the restriction peptide VVVGAVGVGK.In some embodiments, the HLA-peptide antigen is selected from the following: RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV; RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK; RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVGADGVGK; RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGAVGVGK; RAS_G12V MHC class I antigen comprising HLA-A*31:01 and the restriction peptide VVVGAVGVGK; RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVGAVGVGK. MHC class I antigens; RAS_G12V MHC class I antigens comprising HLA-C*01:02 and the restriction peptide AVGVGKSAL; and RAS_G12V MHC class I antigens comprising HLA-A*03:01 and the restriction peptide VVVGAVGVGK. In some embodiments, the HLA-peptide antigens are selected from: RAS_G12C MHC class I antigens comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV; RAS_G12D MHC class I antigens comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK; or RAS_G12V MHC class I antigens comprising HLA-A*11:01 and the restriction peptide VVVGAVGVGK. In some embodiments, the antigen binding protein binds to a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV, wherein the ABP binds to the RAS_G12C MHC class I antigen with higher affinity than an HLA-peptide antigen comprising a different RAS G12 mutation. In some embodiments, the ABP binds to a RAS_G12C MHC class I antigen with higher affinity than an HLA-peptide antigen comprising the restriction peptide KLVVVGAVGV and an HLA-A2 molecule. In some embodiments, the ABP does not bind to an HLA-peptide antigen comprising the restriction peptide KLVVVGAVGV and an HLA-A2 molecule.

[0047] In some embodiments, the HLA-peptide antigen comprises an HLA-restricted peptide that is a peptide fragment of RAS comprising a RAS Q61 mutation. In some embodiments, the Q61 mutation is a Q61H, Q61K, Q61R, or Q61L mutation. In some embodiments, the HLA-peptide antigen is a RAS_Q61H MHC class I antigen comprising HLA-A*01:01 and the restriction peptide ILDTAGHEEY.

[0048] In some embodiments, the HLA-peptide antigen comprises an HLA-restricted peptide that is a peptide fragment of TP53 comprising a TP53 mutation. In some embodiments, the TP53 mutation comprises an R213L, S127Y, Y220C, R175H, or R249M mutation. In some embodiments, the HLA-peptide antigen is a TP53 R213L MHC class I antigen comprising A*02:01 and the restriction peptide YLDDRNTFL.

[0049] In some embodiments, the method includes determining or having determined the presence of any one or more of the HLA-peptide antigen, a peptide of the HLA-peptide antigen, a somatic mutation associated with the HLA-peptide antigen, and an HLA molecule of the HLA-peptide antigen in a biological sample obtained from the subject prior to administering.

[0050] In some aspects, the biological sample is a blood sample or a tumor sample. In some embodiments, the blood sample is a plasma sample or a serum sample.

[0051] In some embodiments, the determining comprises RNASeq, microarray, PCR, nanostring, in situ hybridization (ISH), mass spectrometry, sequencing, or immunohistochemistry (IHC).

[0052] In some embodiments of the method, the method includes determining the presence of an HLA-peptide antigen, peptide, or HLA in a biological sample obtained from the subject, and then administering to the subject an ABP that selectively binds to the HLA-peptide antigen.

[0053] Also provided herein are kits comprising an antigen binding protein disclosed herein or a pharmaceutical composition disclosed herein and instructions for use.

[0054] Also provided herein is a system comprising an isolated HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, and the HLA-peptide antigen is selected from the HLA-peptide antigens set forth in any one of SEQ ID NOs: 10,755 to 29,364; and a phage display library.

[0055] In some embodiments, the HLA-peptide antigen is bound to a solid support. In some embodiments, the solid support comprises a bead, a well, a membrane, a tube, a column, a plate, sepharose, a magnetic bead, a cell, or a chip. In some embodiments, the HLA-peptide antigen comprises a first member of an affinity binding pair, and the solid support comprises a second member of the affinity binding pair. In some embodiments, the first member is streptavidin, and the second member is biotin.

[0056] In some embodiments, the phage display library is a human library. In some embodiments, the phage display library is a humanized library.

[0057] In some embodiments, the system further includes a negative control HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, where the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, and the negative control HLA-peptide antigen comprises a different restricted peptide, a different HLA class I molecule, or a different restricted peptide and a different HLA class I molecule. In some embodiments, the negative control HLA-peptide antigen comprises a different restricted peptide but the same HLA class I molecule as the HLA-peptide antigen.

[0058] In some embodiments, the system comprises a reaction mixture comprising an HLA-peptide antigen and a plurality of phages from a phage display library.

[0059] Also provided herein is the use of the systems disclosed herein to identify antigen binding proteins that selectively bind to isolated HLA-peptide antigens.

[0060] Also provided herein are compositions comprising an HLA-peptide antigen set forth in any one of SEQ ID NOs: 10,755-29,364, wherein the HLA-peptide antigen is covalently linked to an affinity tag. In some embodiments, the affinity tag is a biotin tag.

[0061] Also provided herein are compositions comprising an HLA-peptide antigen set forth in any one of SEQ ID NOS: 10,755-29,364 complexed with a detectable label. In some embodiments, the detectable label comprises a β2-microglobulin binding molecule. In some embodiments, the β2-microglobulin binding molecule is a labeled antibody. In some embodiments, the labeled antibody is a fluorochrome-labeled antibody.

[0062] Also provided herein is a composition comprising an HLA-peptide antigen, wherein the HLA-peptide antigen is set forth in any one of SEQ ID NOS: 10,755-29,364, and is bound to a solid support. In some embodiments, the solid support comprises a bead, a well, a membrane, a tube, a column, a plate, sepharose, a magnetic bead, a cell, or a chip. In some embodiments, the HLA-peptide antigen comprises a first member of an affinity binding pair, and the solid support comprises a second member of the affinity binding pair. In some embodiments, the first member is streptavidin, and the second member is biotin.

[0063] Also provided herein are host cells comprising a heterologous HLA-peptide antigen set forth in any one of SEQ ID NOs: 10,755-29,364. Also provided herein are host cells expressing an HLA subtype defined by any one of the HLA-peptide antigens set forth in SEQ ID NOs: 10,755-29,364. Also provided herein are host cells comprising a polynucleotide encoding an HLA-restricted peptide defined by any one of the HLA-peptide antigens in SEQ ID NOs: 10,755-29,364.

[0064] In some embodiments, the host cells do not contain endogenous MHC. In some embodiments, the host cells contain exogenous HLA. In some embodiments, the host cells are K562 or A375 cells. In some embodiments, the host cells are cultured cells derived from a tumor cell line. In some embodiments, the tumor cell line expresses an HLA subtype defined by the same HLA-peptide antigen as that describing the HLA-restricted peptide. In some embodiments, the tumor cell line is selected from the group consisting of HCC-1599, NCI-H510A, A375, LN229, NCI-H358, ZR-75-1, MS751, OE19, MOR, BV173, MCF-7, NCI-H82, Colo829, SK-MEL-28, KYSE270, 59M, and NCI-H146.

[0065] Also provided herein are cell culture systems and cell culture media comprising the host cells disclosed herein. In some embodiments, the host cells express an HLA subtype defined by any one of the HLA-peptide antigens in SEQ ID NOS: 10,755-21,015 and SEQ ID NOS: 21,016-29,364, and the cell culture media comprises a restriction peptide defined by the same HLA-peptide antigen as the HLA subtype. In some embodiments, the host cells are K562 cells comprising exogenous HLA, the exogenous HLA is an HLA subtype defined by any one of the HLA-peptide antigens in SEQ ID NOS: 10,755-29,364, and the cell culture media comprises a restriction peptide defined by the same HLA-peptide antigen that defines the HLA subtype.

[0066] Also provided herein is a method of identifying an antigen-binding protein disclosed herein, comprising providing at least one HLA-peptide antigen set forth in SEQ ID NOs: 10,755-29,364; and binding at least one target to the antigen-binding protein, thereby identifying the antigen-binding protein.

[0067] In some embodiments, the antigen binding protein is present in a phage display library comprising a plurality of distinct antigen binding proteins, hi some embodiments, the phage display library is substantially free of antigen binding proteins that non-specifically bind to the HLA of the HLA-peptide antigen.

[0068] In some embodiments, the combining step is performed multiple times, optionally at least three times.

[0069] In some embodiments, the method further comprises contacting the antigen binding protein with one or more peptide-HLA complexes distinct from the HLA-peptide antigen to determine whether the antigen binding protein selectively binds to the HLA-peptide antigen, optionally wherein selectivity is determined by measuring the binding affinity of the antigen binding protein to a soluble target HLA-peptide complex and to a soluble HLA-peptide complex distinct from the target complex, optionally wherein selectivity is determined by measuring the binding affinity of the antigen binding protein to a target HLA-peptide complex expressed on the surface of one or more cells and to a HLA-peptide complex distinct from the target complex expressed on the surface of one or more cells.

[0070] Also provided herein is a method for identifying an antigen binding protein disclosed herein, the method comprising obtaining at least one HLA-peptide antigen set forth in SEQ ID NOs: 10,755-29,364; administering the HLA-peptide antigen, optionally in combination with an adjuvant, to a subject; and isolating the antigen binding protein from the subject.

[0071] In some embodiments, isolating the antigen binding protein comprises screening the subject's serum to identify the antigen binding protein.

[0072] In some embodiments, the method further comprises contacting the antigen binding protein with one or more peptide-HLA complexes distinct from the HLA-peptide antigen to determine whether the antigen binding protein selectively binds to the HLA-peptide antigen, optionally wherein selectivity is determined by measuring the binding affinity of the antigen binding protein to the HLA-peptide antigen and to a soluble HLA-peptide complex distinct from the HLA-peptide antigen, optionally wherein selectivity is determined by measuring the binding affinity of the antigen binding protein to an HLA-peptide antigen expressed on the surface of one or more cells and to an HLA-peptide complex distinct from the HLA-peptide antigen expressed on the surface of one or more cells.

[0073] In some embodiments, the subject is a mouse, rabbit, or llama.

[0074] In some embodiments, isolating the antigen binding protein comprises isolating B cells from the subject that express the antigen binding protein, and optionally directly cloning the sequence encoding the antigen binding protein from the isolated B cells. In some embodiments, the method further comprises generating hybridomas using the B cells. In some embodiments, the method further comprises cloning CDRs from the B cells. In some embodiments, the method further comprises immortalizing the B cells, optionally via EBV transformation.

[0075] In some embodiments, the method further comprises generating a library comprising the antigen binding proteins of the B cells, optionally the library is phage display or yeast display.

[0076] In some embodiments, the method further comprises humanizing the antigen binding protein.

[0077] Also provided herein are methods for identifying an antigen-binding protein disclosed herein, comprising obtaining cells containing the antigen-binding protein; contacting the cells with an HLA multimer comprising at least one HLA-peptide antigen set forth in SEQ ID NOs: 10,755-29,364; and identifying the antigen-binding protein via binding between the HLA multimer and the antigen-binding protein. In some embodiments, the method further comprises contacting the cells containing the antigen-binding protein with an HLA multimer comprising a corresponding wild-type sequence of at least one HLA-peptide antigen set forth in SEQ ID NOs: 10,755-29,364, and excluding the antigen-binding protein if it binds to an HLA multimer comprising the corresponding wild-type sequence.

[0078] Also provided herein is a method of identifying an antigen-binding protein disclosed herein, the method comprising providing at least one HLA-peptide antigen set forth in SEQ ID NOs: 10,755-29,364; and identifying the antigen-binding protein using the target.

[0079] Also provided herein is an antigen binding protein (ABP) that specifically binds to an HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, wherein the HLA class I molecule and the HLA-restricted peptide are each selected from the HLA-peptide antigens set forth in any one of SEQ ID NOS: 10,755 to 29,364, and wherein the ABP comprises an alpha CDR3 amino acid sequence and a corresponding beta CDR3 amino acid sequence selected from the group consisting of the sequences set forth in Tables 1C.1, 1C.2, 1C.3, and 1D.

[0080] In some embodiments, the ABP further comprises an alpha variable ("V") segment, an alpha joining ("J") segment, a beta variable ("V") segment, a beta joining ("J") segment, optionally a beta diversity ("D") segment, and optionally a beta constant region selected from the group consisting of the regions set forth in Tables 1C.1, 1C.2, 1C.3, and 1D corresponding to the alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence. In some embodiments, the ABP comprises an alpha variable region and a corresponding beta variable region comprising an amino acid sequence selected from the sequences set forth in Tables 1A.1, 1A.2, 1A.3, and 1B corresponding to the alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence.

[0081] Also provided herein is an antigen binding protein (ABP) that specifically binds to an HLA-peptide antigen comprising an HLA-restricted RAS peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, and wherein the HLA-restricted RAS peptide comprises at least one alteration that distinguishes the HLA-restricted RAS peptide sequence from the corresponding peptide sequence of a wild-type RAS peptide, wherein the ABP comprises an alpha CDR3 amino acid sequence and a corresponding beta CDR3 amino acid sequence selected from the group consisting of the sequences set forth in Tables 1C.1, 1C.2, 1C.3, and 1D.

[0082] In some embodiments, the HLA-peptide antigen is selected from Table 5A. In some embodiments, the HLA-peptide antigen is selected from Table 5B. In some embodiments, the HLA-peptide antigen is selected from Table 6. In some embodiments, the HLA-peptide antigen is selected from Table 7.

[0083] In some embodiments, the HLA-restricted peptide comprises a RAS G12 mutation. In some embodiments, the G12 mutation is a G12C, G12D, G12V, or G12A mutation. In some embodiments, the HLA-peptide antigen comprises an HLA class I molecule selected from HLA-A*02:01, HLA-A*11:01, HLA-A*31:01, HLA-C*01:02, and HLA-A*03:01. In some embodiments, the RAS G12 mutation is any one or more of a KRAS mutation, an NRAS mutation, and an HRAS mutation.

[0084] In some embodiments, the HLA-peptide antigen is a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV. In some embodiments, the ABP comprises an alpha CDR3 amino acid sequence and a corresponding beta CDR3 amino acid sequence selected from the group consisting of the sequences set forth in Table 1C.2. In some embodiments, the ABP further comprises an alpha variable ("V") segment, an alpha joining ("J") segment, a beta variable ("V") segment, a beta joining ("J") segment, optionally a beta diversity ("D") segment, and optionally a beta constant region selected from the group consisting of the regions set forth in Table 1C.2 corresponding to the alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence. In some embodiments, the ABP comprises an alpha variable region and a corresponding beta variable region comprising an amino acid sequence selected from the sequences set forth in Table 1A.2 corresponding to the alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence.

[0085] In some embodiments, the HLA-peptide antigen is a RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVGAVGVGK. In some embodiments, the ABP comprises an alpha CDR3 amino acid sequence and a corresponding beta CDR3 amino acid sequence selected from the group consisting of the sequences set forth in Table 1C.3. In some embodiments, the ABP further comprises an alpha variable ("V") segment, an alpha joining ("J") segment, a beta variable ("V") segment, a beta joining ("J") segment, optionally a beta diversity ("D") segment, and optionally a beta constant region selected from the group consisting of the regions set forth in Table 1C.3 corresponding to the alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence. In some embodiments, the ABP comprises an alpha variable region and a corresponding beta variable region comprising an amino acid sequence selected from the sequences set forth in Table 1A.3 corresponding to the alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence. [The present invention 1001] 1. An antigen-binding protein (ABP) that specifically binds to an HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, wherein the HLA class I molecule and the HLA-restricted peptide are each selected from the HLA-peptide antigens set forth in any one of SEQ ID NOS: 10,755 to 29,364, and the ABP comprises a T cell receptor (TCR) or an antigen-binding fragment thereof. [The present invention 1002] 1001. The ABP of the present invention, wherein said HLA-restricted peptide is about 5 to 15 amino acids in length. [The present invention 1003] 1002. The ABP of the present invention, wherein said HLA-restricted peptide is about 8-12 amino acids in length, optionally 8, 9, 10, 11, or 12 amino acids in length. [The present invention 1004] The HLA-peptide antigen is a. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGADGVGK; b. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK; c. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; d. CTNNB1_S45P MHC class I antigen containing HLA-A*03:01 and the restriction peptide TTAPPLSGK; e. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGADGVGK; f. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGAVGVGK; g. RAS_G12V MHC class I antigen containing HLA-C*01:02 and the restriction peptide AVGVGKSAL; h. RAS_G12V MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGAVGVGK; i. TP53_K132N MHC class I antigen containing HLA-A*24:02 and the restrictive peptide TYSPALNNMF; j.CTNNB1_S37Y MHC class I antigen containing HLA-A*02:01 and the restriction peptide YLDSGIHYGA; k. RAS_G12C MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGACGVGK; l. RAS_G12C MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGACGVGK; m.RAS_G12D MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGADGVGK; n. RAS_Q61H MHC class I antigen containing HLA-A*01:01 and the restriction peptide ILDTAGHEEY; and TP53_R213L MHC class I antigen containing oA*02:01 and the restriction peptide YLDDRNTFL Any of the preceding ABPs of the invention selected from the group consisting of: [The present invention 1005] a. the restriction peptide comprises a RAS_G12A mutation and the HLA class I molecule is HLA-A*02:01; b. the restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-A*02:06; c. The restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*27:05; d. The restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*35:01; e. the restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*41:02; f. The restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*48:01; g. The restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-C*08:03; h. The restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; i. the restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; j. The restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*03:02; k. The restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*68:01; l. The restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-B*27:05; m. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*02:01; n. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*02:05; o. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*03:01; p. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*11:01; q. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*11:01; r. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*26:01; s. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*31:01; t. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*68:01; u. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*07:02; v. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*08:01; w. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*13:02; x. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*15:01; y. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*27:05; z. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*35:01; aa. the restriction peptide comprises the RAS_G12D mutation and the HLA class I molecule is HLA-B*37:01; bb. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*38:01; cc. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*40:01; dd. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*40:02; ee. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*44:02; ff. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*44:03; gg. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*48:01; hh. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*50:01; ii. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*57:01; jj. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*01:02; kk. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*02:02; ll. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*03:03; mm. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*03:04; nn. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*04:01; oo. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*05:01; pp. whether the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*07:04; qq. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*08:02; rr. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*08:03; ss. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*16:01; tt. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*17:01; uu. The restriction peptide contains the RAS_G12R mutation and the HLA class I molecule is HLA-B*41:02; vv. the restriction peptide contains the RAS_G12R mutation and the HLA class I molecule is HLA-C*07:04; ww. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:01; xx. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:05; yy. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:06; zz. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*03:01; aaa. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*03:01; bbb. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*11:01; ccc. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*11:01; ddd. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*25:01; eee. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*26:01; fff. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*30:01; ggg. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*31:01; hhh. The restriction peptide contains the RAS_G12V mutation, and the HLA class I molecule is HLA-A*31:01; iii. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*32:01; jjj. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*68:02; kkk. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*07:02; III. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*08:01; mmm. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*13:02; nnn. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*14:02; ooo. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*15:01; ppp. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*27:05; qqq. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*39:01; rrr. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*40:01; sss. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*40:02; ttt. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*41:02; uuu. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*44:05; vvv. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*50:01; www. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*51:01; xxx. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*01:02; yyy. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*01:02; zzz. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*03:03; aaaa. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*03:04; bbbb. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*08:02; cccc. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*14:02; dddd. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*17:01; eeee. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-A*02:01; ffff. the restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*07:02; gggg. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*08:01; hhhh. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:01; iiii. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:03; jjjj. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:08; kkkk. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*38:01; III. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-C*04:01; mmmm. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*01:01; nnnn. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*02:01; oooo. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*23:01; pppp. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*29:01; qqqq. Whether the restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*30:02; rrrr. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*33:01; ssss. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*68:01; tttt. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*07:02; uuuu. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*08:01; vvvv. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*18:01; wwww. the restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*35:01; xxxx. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*38:01; yyyy. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*40:01; zzzz. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*44:02; aaaaa. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-C*03:04; bbbbb. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-C*05:01; or ccccc. The restriction peptide contains the KRAS_Q61H mutation, and the HLA class I molecule is HLA-C*08:02. Any of ABPs 1001 to 1003 of the present invention. [The present invention 1006] a. the restricted peptide contains a KRAS_G13D mutation and the HLA class I molecule is C*08:02 or A*11:01; b. The restriction peptide contains the KRAS_Q61K mutation and the HLA class I molecule is A*01:01; c. The restriction peptide contains the NRAS_Q61K mutation and the HLA class I molecule is A*01:01; d. The restriction peptide comprises the TP53_R249M mutation and the HLA class I molecule is B*35:12, B*35:03, or B*35:01; e. the restriction peptide comprises the CTNNB1_S45P mutation and the HLA class I molecule is A*03:01, A*11:01, A*68:01, or A*03:02; f. the restriction peptide comprises the CTNNB1_S45F mutation and the HLA class I molecule is A*03:01, A*11:01, or A*68:01; g. The restriction peptide comprises the ERBB2_Y772_A775dup mutation and the HLA class I molecule is B*18:01; h. the restricted peptide contains a KRAS_G12D mutation and the HLA class I molecule is A*11:01, A*03:01, or C*08:02; i. the restriction peptide contains the NRAS_G12D mutation and the HLA class I molecule is A*11:01, A*03:01, or C*08:02; j. The restriction peptide contains the KRAS_Q61R mutation and the HLA class I molecule is A*01:01; k. The restriction peptide contains the NRAS_Q61R mutation and the HLA class I molecule is A*01:01; l. the restriction peptide comprises the CTNNB1_T41A mutation and the HLA class I molecule is A*03:01, A*03:02, A*11:01, B*15:10, C*03:03, or C*03:04; m. The restriction peptide contains the TP53_K132N mutation and the HLA class I molecule is A*24:02 or A*23:01; n. The restriction peptide contains the KRAS_G12A mutation and the HLA class I molecule is A*03:01 or A*11:01; o. The restriction peptide contains the KRAS_Q61L mutation and the HLA class I molecule is A*01:01; p. the restriction peptide contains the NRAS_Q61L mutation and the HLA class I molecule is A*01:01; q. the restriction peptide comprises the TP53_R213L mutation and the HLA class I molecule is A*02:07, C*08:02, or A*02:01; r. The restriction peptide contains the BRAF_G466V mutation, and the HLA class I molecule is B*15:01 or B*15:03; s. the restriction peptide contains a KRAS_G12V mutation and the HLA class I molecule is A*03:01, A*03:02, A*11:01, or C*01:02; t. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is A*01:01; u. The restriction peptide contains the NRAS_Q61H mutation and the HLA class I molecule is A*01:01; v. the restriction peptide comprises the CTNNB1_S37F mutation and the HLA class I molecule is A*01:01, A*23:01, A*24:02, B*15:10, B*39:06, C*05:01, C*14:02, or C*14:03; w. the restriction peptide contains the TP53_S127Y mutation and the HLA class I molecule is A*11:01 or A*03:01; x. the restriction peptide contains the TP53_K132E mutation and the HLA class I molecule is A*24:02, C*14:03, or A*23:01; y. the restricted peptide contains a KRAS_G12C mutation and the HLA class I molecule is A*02:01, A*11:01, or A*03:01; z. the restriction peptide contains the NRAS_G12C mutation and the HLA class I molecule is A*02:01, A*11:01, or A*03:01; aa. The restricted peptide contains the EGFR_L858R mutation and the HLA class I molecule is A*11:01 or A*03:01; bb. the restriction peptide comprises the TP53_Y220C mutation and the HLA class I molecule is A*02:01; or cc. The restriction peptide comprises the TP53_R175H mutation, and the HLA class I molecule is A*02:01; Any of ABPs 1001 to 1003 of the present invention. [The present invention 1007] The HLA-peptide antigen is CTNNB1_S45P MHC class I antigen containing aA*11:01 and the constraining peptide TTAPPLSGK; CTNNB1_T41AMHC class I antigen containing bA*11:01 and the constraint peptide ATAPSLSGK; RAS_G12D MHC class I antigen containing cA*11:01 and the constraint peptide VVVGADGVGK; RAS_G12V MHC class I antigen containing dA*03:01 and the constraint peptide VVGAVGVGK; RAS_G12V MHC class I antigen containing eA*03:01 and the constraint peptide VVVGAVGVGK; RAS_G12V MHC class I antigen containing fA*11:01 and the constraint peptide VVGAVGVGK; RAS_G12V MHC class I antigen containing gA*11:01 and the constraint peptide VVVGAVGVGK; KRAS_Q61R MHC class I antigen comprising hA*01:01 and the constraint peptide ILDTAGREEY; and TP53_R213L MHC class I antigen containing iA*02:01 and the restriction peptide YLDDRNTFL Any of ABPs 1001 to 1003 of the present invention selected from the following. [The present invention 1008] The ABP of any one of 1001 to 1003, wherein the HLA-restricted peptide comprises a RAS G12 mutation. [The present invention 1009] 1008. The ABP of the present invention, wherein the G12 mutation is a G12C, G12D, G12V, or G12A mutation. [The present invention 1010] 1008. The ABP of the present invention, wherein the HLA-peptide antigen comprises an HLA class I molecule selected from HLA-A*02:01, HLA-A*11:01, HLA-A*31:01, HLA-C*01:02, and HLA-A*03:01. [The present invention 1011] The ABP of any one of 1008 to 1010, wherein the RAS G12 mutation is any one or more of a KRAS mutation, an NRAS mutation, and an HRAS mutation. [The present invention 1012] The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12C MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGACGVGK; c. RAS_G12C MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGACGVGK; d. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGADGVGK; e. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGADGVGK; f. RAS_G12D MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGADGVGK; g. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK; h. RAS_G12V MHC class I antigen containing HLA-A*31:01 and the restriction peptide VVVGAVGVGK; i. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGAVGVGK; j. RAS_G12V MHC class I antigen containing HLA-C*01:02 and the restriction peptide AVGVGKSAL; and k. RAS_G12V MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGAVGVGK 1009 ABPs of the present invention selected from: [The present invention 1013] The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGADGVGK; c. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGADGVGK; d. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK; e. RAS_G12V MHC class I antigen containing HLA-A*31:01 and the restriction peptide VVVGAVGVGK; f. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGAVGVGK; g. RAS_G12V MHC class I antigen containing HLA-C*01:02 and the restriction peptide AVGVGKSAL; and h. RAS_G12V MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGAVGVGK 1009 ABPs of the present invention selected from: [The present invention 1014] The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGADGVGK; and c. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK 1009 ABPs of the present invention selected from: [The present invention 1015] 1009. The ABP of the present invention, wherein said HLA-peptide antigen is a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV. [The present invention 1016] 1009. The ABP of the present invention, wherein said HLA-peptide antigen is a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK. [The present invention 1017] 1009. The ABP of the present invention, wherein said HLA-peptide antigen is a RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGAVGVGK. [The present invention 1018] 1004. The ABP of any one of claims 1001 to 1003, wherein the HLA-restricted peptide comprises a RAS Q61 mutation. [The present invention 1019] 1018. The ABP of the present invention, wherein the Q61 mutation is a Q61H, Q61K, Q61R, or Q61L mutation. [The present invention 1020] 1018. The ABP of the present invention, wherein the HLA-peptide antigen is a RAS_Q61H MHC class I antigen comprising HLA-A*01:01 and the restriction peptide ILDTAGHEEY. [The present invention 1021] 1004. The ABP of any one of claims 1001 to 1003, wherein the HLA-restricted peptide comprises a TP53 mutation. [The present invention 1022] 1021. The ABP of the present invention, wherein the TP53 mutation comprises a R213L, S127Y, Y220C, R175H, or R249M mutation. [The present invention 1023] 1021. The ABP of the present invention, wherein said HLA-peptide antigen is a TP53 R213L MHC class I antigen comprising A*02:01 and the restriction peptide YLDDRNTFL. [The present invention 1024] Any of the preceding ABPs of the invention, which bind to the HLA-peptide antigen through at least one contact point with the HLA class I molecule and through at least one contact point with the HLA-restricted peptide. [The present invention 1025] 10. The ABP of any of the prior inventions, wherein said antigen binding protein binds to a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV, and wherein said ABP binds to said RAS_G12C MHC class I antigen with higher affinity than an HLA-peptide antigen comprising a different RAS G12 mutation. [The present invention 1026] 1025 ABP of the present invention, which binds to said RAS_G12C MHC class I antigen with higher affinity than an HLA-peptide antigen comprising the restriction peptide KLVVVGAVGV and an HLA-A2 molecule. [The present invention 1027] 1026 ABP of the present invention, which does not bind to HLA-peptide antigens containing the restriction peptide KLVVVGAVGV and HLA-A2 molecules. [The present invention 1028]

[0023] Any of the preceding ABPs of the invention, wherein said antigen binding protein is linked to a scaffold, optionally said scaffold comprising serum albumin or Fc, optionally wherein Fc is human Fc and is an IgG (IgG1, IgG2, IgG3, IgG4), IgA (IgA1, IgA2), IgD, IgE, or IgM isotype Fc. [The present invention 1029] Any of the prior ABPs of the invention, wherein said antigen binding protein is linked to a scaffold via a linker, optionally said linker is a peptide linker, optionally said peptide linker is a hinge region of a human antibody. [The present invention 1030] Any of the preceding ABPs of the invention, wherein said TCR or antigen binding portion thereof comprises a TCR variable region. [The present invention 1031] Any of the preceding ABPs of the invention, wherein said TCR or antigen binding portion thereof comprises one or more TCR complementarity determining regions (CDRs). [The present invention 1032] Any of the prior ABPs of the invention, wherein said TCR comprises an alpha chain and a beta chain. [The present invention 1033] Any of the prior ABPs of the invention, wherein said TCR comprises a gamma chain and a delta chain. [The present invention 1034] Any of the prior ABPs of the invention, wherein said TCR comprises a single chain TCR (scTCR). [This invention 1035] Any of the preceding ABPs of the invention, wherein said TCR comprises a recombinant TCR sequence. [The present invention 1036] Any of the prior ABPs of the invention, wherein said TCR comprises a human TCR sequence, and optionally said human TCR sequence is a fully human TCR sequence. [This invention 1037] Any of the preceding ABPs of the invention, wherein said TCR comprises a modified TCR alpha constant (TRAC) region, a modified TCR beta constant (TRBC) region, or a modified TRAC region and a modified TRBC region. [The present invention 1038] Any of the preceding ABPs of the present invention comprising a modification that extends its half-life. [This invention 1039]

[0023] Any of the preceding ABPs of the present invention, which is part of a chimeric antigen receptor (CAR) comprising an extracellular portion comprising an antigen binding protein and an intracellular signaling domain. [The present invention 1040] The ABP of the present invention, wherein the intracellular signaling domain comprises an ITAM. [The present invention 1041] The ABP of the present invention 1039 or 1040, wherein the intracellular signaling domain comprises the signaling domain of the zeta chain of the CD3-zeta (CD3) chain. [The present invention 1042] The antigen-binding protein of any of claims 1039 to 1041, further comprising a transmembrane domain linking said extracellular domain and said intracellular signaling domain. [This invention 1043] 1042. The ABP of the present invention, wherein the transmembrane domain comprises the transmembrane portion of CD28. [This invention 1044] The antigen-binding protein of any of claims 1039 to 1043, further comprising an intracellular signaling domain of a T cell costimulatory molecule. [This invention 1045] The ABP of the present invention 1044, wherein said T cell costimulatory molecule is CD28, 4-1BB, OX-40, ICOS, or any combination thereof. [The present invention 1046] Any ABP of the preceding invention for use as a medicament. [This invention 1047] The ABP of any of the preceding inventions for use in treating said cancer, optionally wherein said cancer expresses or is predicted to express said HLA-peptide antigen. [This invention 1048] The ABP of any of the preceding inventions for use in treating cancer, wherein the cancer is selected from a solid tumor and a hematological tumor. [This invention 1049] An antigen binding protein (ABP) that competes for binding with any of the preceding ABPs of the invention. [The present invention 1050] An antigen binding protein (ABP) that binds to the same HLA-peptide antigen epitope as that bound by any of the preceding ABPs of the invention. [This invention 1051] An engineered cell expressing a receptor comprising any of the antigen binding proteins of the preceding invention. [This invention 1052] The engineered cells of the present invention 1051 are T cells. [This invention 1053] 1052. The engineered cell of the present invention, wherein the T cells are selected from the group consisting of naive T (TN) cells, effector T cells (TEFF), memory T cells, stem cell memory T cells (TSCM), central memory T cells (TCM), effector memory T cells (TEM), terminally differentiated effector memory T cells, tumor infiltrating lymphocytes (TIL), immature T cells, mature T cells, helper T cells, cytotoxic T cells, mucosal-associated invariant T (MALT) cells, regulatory T cells (Treg), TH1 cells, TH2 cells, TH3 cells, TH17 cells, TH9 cells, TH22 cells, follicular helper T cells, natural killer T cells (NKT), alpha-beta T cells, and gamma-delta T cells. [This invention 1054] The engineered cell of claim 1052, wherein said T cells are cytotoxic T cells (CTLs). [This invention 1055] The engineered cell of any one of claims 1051 to 1054, which is a human cell or a human-derived cell. [The present invention 1056] The engineered cell of any one of 1051 to 1055 of the present invention, which is an autologous cell of the subject. [This invention 1057] The engineered cell of claim 1056, wherein said subject is known to have or suspected of having cancer. [This invention 1058] The engineered cell of any one of claims 1056 to 1057, wherein the autologous cell is a cell isolated from the subject. [This invention 1059] 1058. The engineered cell of claim 1058, wherein said isolated cells are ex vivo cultured cells, and optionally said ex vivo cultured cells are stimulated cells. [The present invention 1060] The engineered cell of any one of claims 1056 to 1057, wherein the autologous cell is an in vivo engineered cell. [The present invention 1061] 1060. The engineered cell of any of claims 1051 to 1060, wherein said antigen binding protein is expressed by a heterologous promoter. [The present invention 1062] 1061. The engineered cell of any of claims 1051 to 1061, wherein the ABP comprises a T cell receptor (TCR) or an antigen-binding portion thereof, and a polynucleotide encoding the T cell receptor (TCR) or an antigen-binding portion thereof is inserted into an endogenous TCR locus. [This invention 1063] The engineered cell of any of claims 1051 to 1062, which does not express endogenous ABP. [This invention 1064] An isolated polynucleotide encoding any of the preceding ABPs of the invention or antigen-binding portions thereof, or a set of polynucleotides encoding any of the preceding ABPs of the invention or antigen-binding portions thereof. [This invention 1065] A vector or set of vectors comprising a polynucleotide or set of polynucleotides of the present invention 1064. [The present invention 1066] A virus comprising an isolated polynucleotide or set of polynucleotides of the present invention. [This invention 1067] 1066. The virus of the present invention, which is a filamentous phage. [The present invention 1068] A yeast cell comprising any of the isolated polynucleotides or sets of polynucleotides of the preceding invention. [This invention 1069] A host cell comprising a polynucleotide or set of polynucleotides of any of the preceding inventions or a vector or set of vectors of the invention 1065, and which is optionally a CHO or HEK293 or optionally a T cell. [The present invention 1070] A method of producing an antigen binding protein, comprising expressing said antigen binding protein using the host cell of the invention 1069 and isolating said expressed antigen binding protein. [This invention 1071] A pharmaceutical composition comprising any of the preceding antigen binding proteins of the invention and a pharmaceutically acceptable excipient. [This invention 1072] A method of treating cancer in a subject, comprising administering to said subject any of the ABPs of the preceding inventions, any of the engineered cells of inventions 1051-1063, or the pharmaceutical composition of invention 1071, optionally wherein said cancer is selected from solid tumors and hematological tumors. [This invention 1073] A method of stimulating an immune response in a subject, comprising administering to said subject any of the ABPs of the preceding inventions, any of the engineered cells of inventions 1051-1063, or the pharmaceutical composition of invention 1071, optionally wherein said subject has cancer, and optionally wherein said cancer is selected from a solid tumor and a hematological tumor. [This invention 1074] A method of killing target cells in a subject, comprising administering to said subject any of the ABPs of the preceding inventions, any of the engineered cells of inventions 1051-1063, or the pharmaceutical composition of invention 1071, optionally wherein said subject has cancer and said target cells are cancer cells, optionally wherein said cancer is selected from a solid tumor and a hematological tumor. [This invention 1075] 1075. The method of any one of claims 1072 to 1074, wherein the subject is a human subject. [This invention 1076] Any of the methods of claims 1072 to 1074, wherein the cancer expresses or is predicted to express an HLA-peptide antigen or an HLA class I molecule set forth in any one of SEQ ID NOs: 10,755 to 29,364, and the ABP binds to the HLA-peptide antigen. [This invention 1077] Any of the methods of claims 1072 to 1074, wherein the cancer expresses or is predicted to express an HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, the HLA-restricted peptide being located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, wherein the HLA class I molecule and the HLA-restricted peptide are each selected from the HLA-peptide antigens set forth in any one of SEQ ID NOs: 10,755 to 29,364, and wherein the ABP binds to the HLA-peptide antigen. [This invention 1078] The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGADGVGK; c. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK; d. CTNNB1_S45P MHC class I antigen containing HLA-A*03:01 and the restriction peptide TTAPPLSGK; e. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGADGVGK; f. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGAVGVGK; g. RAS_G12V MHC class I antigen containing HLA-C*01:02 and the restriction peptide AVGVGKSAL; h. RAS_G12V MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGAVGVGK; i. TP53_K132N MHC class I antigen containing HLA-A*24:02 and the restrictive peptide TYSPALNNMF; j.CTNNB1_S37Y MHC class I antigen containing HLA-A*02:01 and the restriction peptide YLDSGIHYGA; k. RAS_G12C MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGACGVGK; l. RAS_G12C MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGACGVGK; m.RAS_G12D MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGADGVGK; n. RAS_Q61H MHC class I antigen containing HLA-A*01:01 and the restriction peptide ILDTAGHEEY; and TP53_R213L MHC class I antigen containing oA*02:01 and the restriction peptide YLDDRNTFL 1077. The method of claim 1077, wherein the method is selected from the group consisting of: [This invention 1079] a. the restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-A*02:01; b. the restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-A*02:06; c. The restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*27:05; d. The restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*35:01; e. the restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*41:02; f. The restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*48:01; g. The restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-C*08:03; h. The restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; i. the restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; j. The restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*03:01; k. The restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*03:02; l. The restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*68:01; m. The restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-B*27:05; n. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*02:01; o. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*02:05; p. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*03:01; q. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*11:01; r. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*11:01; s. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*26:01; t. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*31:01; u. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*68:01; v. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*07:02; w. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*08:01; x. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*13:02; y. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*15:01; z. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*27:05; aa. the restriction peptide comprises the RAS_G12D mutation and the HLA class I molecule is HLA-B*35:01; bb. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*37:01; cc. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*38:01; dd. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*40:01; ee. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*40:02; ff. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*44:02; gg. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*44:03; hh. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*48:01; ii. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*50:01; jj. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*57:01; kk. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*01:02; ll. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*02:02; mm. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*03:03; nn. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*03:04; oo. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*04:01; pp. whether the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*05:01; qq. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*07:04; rr. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*08:02; ss. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*08:03; tt. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*16:01; uu. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*17:01; vv. the restriction peptide contains the RAS_G12R mutation and the HLA class I molecule is HLA-B*41:02; ww. the restriction peptide contains the RAS_G12R mutation and the HLA class I molecule is HLA-C*07:04; xx. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:01; yy. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:05; zz. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:06; aaa. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*03:01; bbb. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*03:01; ccc. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*11:01; ddd. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*11:01; eee. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*25:01; fff. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*26:01; ggg. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*30:01; hhh. The restriction peptide contains the RAS_G12V mutation, and the HLA class I molecule is HLA-A*31:01; iii. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*31:01; jjj. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*32:01; kkk. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*68:02; III. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*07:02; mmm. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*08:01; nnn. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*13:02; ooo. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*14:02; ppp. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*15:01; qqq. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*27:05; rrr. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*39:01; sss. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*40:01; ttt. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*40:02; uuu. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*41:02; vvv. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*44:05; www. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*50:01; xxx. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*51:01; yyy. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*01:02; zzz. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*01:02; aaaa. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*03:03; bbbb. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*03:04; cccc. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*08:02; dddd. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*14:02; eeee. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*17:01; ffff. the restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-A*02:01; gggg. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*07:02; hhhh. The restriction peptide contains the KRAS_G13D mutation, and the HLA class I molecule is HLA-B*08:01; iiii. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:01; jjjj. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:03; kkkk. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:08; III. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*38:01; mmmm. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-C*04:01; nnnn. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*01:01; oooo. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*02:01; pppp. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*23:01; qqqq. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*29:01; rrrr. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*30:02; ssss. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*33:01; tttt. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*68:01; uuuu. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*07:02; vvvv. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*08:01; wwww. the restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*18:01; xxxx. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*35:01; yyyy. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*38:01; zzzz. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*40:01; aaaaa. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*44:02; bbbbb. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-C*03:04; ccccc. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-C*05:01; or ddddd. The restriction peptide contains the KRAS_Q61H mutation, and the HLA class I molecule is HLA-C*08:02; The method of the present invention 1077. [The present invention 1080] a. the restricted peptide contains a KRAS_G13D mutation and the HLA class I molecule is C*08:02 or A*11:01; b. The restriction peptide contains the KRAS_Q61K mutation and the HLA class I molecule is A*01:01; c. The restriction peptide contains the NRAS_Q61K mutation and the HLA class I molecule is A*01:01; d. The restriction peptide comprises the TP53_R249M mutation and the HLA class I molecule is B*35:12, B*35:03, or B*35:01; e. the restriction peptide comprises the CTNNB1_S45P mutation and the HLA class I molecule is A*03:01, A*11:01, A*68:01, or A*03:02; f. the restriction peptide comprises the CTNNB1_S45F mutation and the HLA class I molecule is A*03:01, A*11:01, or A*68:01; g. The restriction peptide comprises the ERBB2_Y772_A775dup mutation and the HLA class I molecule is B*18:01; h. the restricted peptide contains a KRAS_G12D mutation and the HLA class I molecule is A*11:01, A*03:01, or C*08:02; i. the restriction peptide contains the NRAS_G12D mutation and the HLA class I molecule is A*11:01, A*03:01, or C*08:02; j. The restriction peptide contains the KRAS_Q61R mutation and the HLA class I molecule is A*01:01; k. The restriction peptide contains the NRAS_Q61R mutation and the HLA class I molecule is A*01:01; l. the restriction peptide comprises the CTNNB1_T41A mutation and the HLA class I molecule is A*03:01, A*0302, A*11:01, B*15:10, C*03:03, or C*03:04; m. The restriction peptide contains the TP53_K132N mutation and the HLA class I molecule is A*24:02 or A*23:01; n. The restriction peptide contains the KRAS_G12A mutation and the HLA class I molecule is A*03:01 or A*11:01; o. The restriction peptide contains the KRAS_Q61L mutation and the HLA class I molecule is A*01:01; p. the restriction peptide contains the NRAS_Q61L mutation and the HLA class I molecule is A*01:01; q. the restriction peptide comprises the TP53_R213L mutation and the HLA class I molecule is A*02:07, C*08:02, or A*02:01; r. The restriction peptide contains the BRAF_G466V mutation, and the HLA class I molecule is B*15:01 or B*15:03; s. the restriction peptide contains a KRAS_G12V mutation and the HLA class I molecule is A*03:01, A*03:02, A*11:01, or C*01:02; t. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is A*01:01; u. The restriction peptide contains the NRAS_Q61H mutation and the HLA class I molecule is A*01:01; v. the restriction peptide comprises the CTNNB1_S37F mutation and the HLA class I molecule is A*01:01, A*23:01, A*24:02, B*15:10, B*39:06, C*05:01, C*14:02, or C*14:03; w. the restriction peptide contains the TP53_S127Y mutation and the HLA class I molecule is A*11:01 or A*03:01; x. the restriction peptide contains the TP53_K132E mutation and the HLA class I molecule is A*24:02, C*14:03, or A*23:01; y. the restricted peptide contains a KRAS_G12C mutation and the HLA class I molecule is A*02:01, A*11:01, or A*03:01; z. the restriction peptide contains the NRAS_G12C mutation and the HLA class I molecule is A*02:01, A*11:01, or A*03:01; aa. The restricted peptide contains the EGFR_L858R mutation and the HLA class I molecule is A*11:01 or A*03:01; bb. the restriction peptide comprises the TP53_Y220C mutation and the HLA class I molecule is A*02:01; or cc. The restriction peptide comprises the TP53_R175H mutation, and the HLA class I molecule is A*02:01; The method of the present invention 1077. [This invention 1081] The HLA-peptide antigen is CTNNB1_S45P MHC class I antigen containing aA*11:01 and the constraining peptide TTAPPLSGK; CTNNB1_T41A MHC class I antigen containing bA*11:01 and the constraint peptide ATAPSLSGK; RAS_G12D MHC class I antigen containing cA*11:01 and the constraint peptide VVVGADGVGK; RAS_G12V MHC class I antigen containing dA*03:01 and the constraint peptide VVGAVGVGK; RAS_G12V MHC class I antigen containing eA*03:01 and the constraint peptide VVVGAVGVGK; RAS_G12V MHC class I antigen containing fA*11:01 and the constraint peptide VVGAVGVGK; RAS_G12V MHC class I antigen containing gA*11:01 and the constraint peptide VVVGAVGVGK; KRAS_Q61R MHC class I antigen comprising hA*01:01 and the constraint peptide ILDTAGREEY; and TP53_R213L MHC class I antigen containing iA*02:01 and the restriction peptide YLDDRNTFL 1077. The method of claim 1077, wherein the method is selected from the group consisting of: [This invention 1082] 1078. The method of claim 1077, wherein said HLA-peptide antigen comprises an HLA-restricted peptide that is a peptide fragment of RAS containing the RAS G12 mutation. [This invention 1083] 1083. The method of claim 1082, wherein said G12 mutation is a G12C, G12D, G12V, or G12A mutation. [This invention 1084] 1082. The method of claim 1082, wherein the HLA-peptide antigen comprises an HLA class I molecule selected from HLA-A*02:01, HLA-A*11:01, HLA-A*31:01, HLA-C*01:02, and HLA-A*03:01. [This invention 1085] The method of any one of claims 1082 to 1084, wherein the RAS G12 mutation is any one or more of a KRAS mutation, an NRAS mutation, and an HRAS mutation. [The present invention 1086] The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12C MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGACGVGK; c. RAS_G12C MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGACGVGK; d. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGADGVGK; e. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGADGVGK; f. RAS_G12D MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGADGVGK; g. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK; h. RAS_G12V MHC class I antigen containing HLA-A*31:01 and the restriction peptide VVVGAVGVGK; i. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGAVGVGK; j. RAS_G12V MHC class I antigen containing HLA-C*01:02 and the restriction peptide AVGVGKSAL; and k. RAS_G12V MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGAVGVGK The method of the present invention 1083, selected from the following. [This invention 1087] The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGADGVGK; c. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGADGVGK; d. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK; e. RAS_G12V MHC class I antigen containing HLA-A*31:01 and the restriction peptide VVVGAVGVGK; f. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGAVGVGK; g. RAS_G12V MHC class I antigen containing HLA-C*01:02 and the restriction peptide AVGVGKSAL; and h. RAS_G12V MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGAVGVGK The method of the present invention 1083, selected from the following. [This invention 1088] The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGADGVGK; or c. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK The method of the present invention 1083, selected from the following. [This invention 1089] 1083. The method of claim 1083, wherein said antigen binding protein binds to a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV, and said ABP binds to said RAS_G12C MHC class I antigen with higher affinity than an HLA-peptide antigen comprising a different RAS G12 mutation. [The present invention 1090] 1089. The method of claim 1089, wherein said ABP binds to said RAS_G12C MHC class I antigen with higher affinity than an HLA-peptide antigen comprising the restriction peptide KLVVVGAVGV and an HLA-A2 molecule. [This invention 1091] 1089. The method of claim 1089, wherein said ABP does not bind to an HLA-peptide antigen comprising the restriction peptide KLVVVGAVGV and an HLA-A2 molecule. [This invention 1092] 1078. The method of claim 1077, wherein said HLA-peptide antigen comprises an HLA-restricted peptide that is a peptide fragment of RAS containing the RAS Q61 mutation. [This invention 1093] 1092. The method of claim 1092, wherein the Q61 mutation is a Q61H, Q61K, Q61R, or Q61L mutation. [This invention 1094] 1092. The method of claim 1092, wherein said HLA-peptide antigen is a RAS_Q61H MHC class I antigen comprising HLA-A*01:01 and the restriction peptide ILDTAGHEEY. [This invention 1095] 1078. The method of claim 1077, wherein said HLA-peptide antigen comprises an HLA-restricted peptide that is a peptide fragment of TP53 comprising a TP53 mutation. [This invention 1096] 1095. The method of claim 1095, wherein the TP53 mutation comprises a R213L, S127Y, Y220C, R175H, or R249M mutation. [This invention 1097] 1095. The method of claim 1095, wherein said HLA-peptide antigen is a TP53 R213L MHC class I antigen comprising A*02:01 and the restriction peptide YLDDRNTFL. [This invention 1098] Any of the methods of claims 1072 to 1097, comprising determining or having determined the presence of any one or more of the HLA-peptide antigen, a peptide of the HLA-peptide antigen, a somatic mutation associated with the HLA-peptide antigen, and an HLA molecule of the HLA-peptide antigen in a biological sample obtained from the subject prior to said administering. [This invention 1099] 1098. The method of claim 8, wherein said biological sample is a blood sample or a tumor sample. [The present invention 1100] 1099. The method of claim 1099, wherein said blood sample is a plasma sample or a serum sample. [The present invention 1101] 1098. The method of claim 1098, wherein said determining comprises RNASeq, microarray, PCR, nanostring, in situ hybridization (ISH), mass spectrometry, sequencing, or immunohistochemistry (IHC). [The present invention 1102] 1098. The method of claim 1098, wherein after determining the presence of the HLA-peptide antigen, peptide, or HLA in the biological sample obtained from the subject, an ABP that selectively binds to the HLA-peptide antigen is administered to the subject. [The present invention 1103] A kit comprising any of the preceding antigen binding proteins of the invention or a pharmaceutical composition of the invention 1071 and instructions for use. [The present invention 1104] a. an isolated HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, and the HLA-peptide antigen is selected from the HLA-peptide antigens set forth in any one of SEQ ID NOs: 10,755 to 29,364; b. Phage display libraries and Including, the system. [This invention 1105] 1104. The system of claim 1104, wherein said HLA-peptide antigen is bound to a solid support. [The present invention 1106] The system of the present invention 1105, wherein said solid support comprises a bead, a well, a membrane, a tube, a column, a plate, sepharose, a magnetic bead, a cell, or a chip. [This invention 1107] 1107. The system of claim 1105 or 1106, wherein said HLA-peptide antigen comprises a first member of an affinity binding pair and said solid support comprises a second member of said affinity binding pair. [This invention 1108] The system of claim 1107, wherein said first member is streptavidin and said second member is biotin. [This invention 1109] The system of any one of 1104 to 1108, wherein the phage display library is a human library. [The present invention 1110] The system of any one of 1104 to 1108, wherein the phage display library is a humanized library. [The present invention 1111] Any of the systems of the present inventions 1104 to 1110, further comprising a negative control HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, and the negative control HLA-peptide antigen comprises a different restricted peptide, a different HLA class I molecule, or a different restricted peptide and a different HLA class I molecule. [The present invention 1112] 1111. The system of claim 1111, wherein the negative control HLA-peptide antigen comprises a different restriction peptide but the same HLA class I molecule as the HLA-peptide antigen. [The present invention 1113] The system of any one of claims 1104 to 1112, comprising a reaction mixture, said reaction mixture comprising said HLA-peptide antigen and a plurality of phages derived from said phage display library. [This invention 1114] Use of the system of any of claims 1104 to 1113 for identifying an antigen-binding protein that selectively binds to the isolated HLA-peptide antigen. [This invention 1115] A composition comprising an HLA-peptide antigen set forth by any one of SEQ ID NOs: 10,755 to 29,364, wherein the HLA-peptide antigen is covalently linked to an affinity tag. [The present invention 1116] 1115. The composition of claim 1115, wherein said affinity tag is a biotin tag. [This invention 1117] A composition comprising an HLA-peptide antigen set forth by any one of SEQ ID NOs: 10,755 to 29,364 complexed to a detectable label. [This invention 1118] 1117. The composition of claim 1117, wherein said detectable label comprises a β2-microglobulin binding molecule. [This invention 1119] 1118. The composition of claim 1118, wherein the β2-microglobulin binding molecule is a labeled antibody. [The present invention 1120] The composition of the present invention 1119, wherein the labeled antibody is a fluorescent dye-labeled antibody. [This invention 1121] A composition comprising an HLA-peptide antigen, wherein the HLA-peptide antigen is set forth in any one of SEQ ID NOs: 10,755 to 29,364, and is bound to a solid support. [This invention 1122] 1121. The composition of claim 1121, wherein said solid support comprises a bead, a well, a membrane, a tube, a column, a plate, sepharose, a magnetic bead, a cell, or a chip. [This invention 1123] 1123. The composition of claim 1121 or 1122, wherein said HLA-peptide antigen comprises a first member of an affinity binding pair and said solid support comprises a second member of said affinity binding pair. [This invention 1124] 1124. The composition of claim 1123, wherein said first member is streptavidin and said second member is biotin. [This invention 1125] A host cell comprising a heterologous HLA-peptide antigen as set forth in any one of SEQ ID NOs: 10,755 to 29,364. [The present invention 1126] A host cell that expresses an HLA subtype defined by any one of the HLA-peptide antigens set forth in SEQ ID NOs: 10,755 to 29,364. [This invention 1127] A host cell comprising a polynucleotide encoding an HLA-restricted peptide defined by any one of the HLA-peptide antigens in SEQ ID NOs: 10,755 to 29,364. [This invention 1128] A host cell of the present invention 1127 that does not contain endogenous MHC. [This invention 1129] A host cell of the present invention 1128 containing exogenous HLA. [The present invention 1130] The host cell of the present invention 1129, which is a K562 or A375 cell. [This invention 1131] The host cell of any one of 1125 to 1130 of the present invention, which is a cultured cell derived from a tumor cell line. [This invention 1132] The host cell of the present invention 1131, wherein said tumor cell line expresses an HLA subtype defined by the same HLA-peptide antigen as that described for the HLA-restricted peptide of the present invention 1127. [This invention 1133] The host cell of the present invention 1131, wherein the tumor cell line is selected from the group consisting of HCC-1599, NCI-H510A, A375, LN229, NCI-H358, ZR-75-1, MS751, OE19, MOR, BV173, MCF-7, NCI-H82, Colo829, SK-MEL-28, KYSE270, 59M, and NCI-H146. [This invention 1134] a. A host cell according to any one of claims 1125 to 1133 of the present invention; b. Cell culture medium and A cell culture system comprising: [This invention 1135] The cell culture system of the present invention 1134, wherein the host cells express an HLA subtype defined by any one of the HLA-peptide antigens in SEQ ID NOs: 10,755 to 21,015 and SEQ ID NOs: 21,016 to 29,364, and the cell culture medium contains a restriction peptide defined by the same HLA-peptide antigen as the HLA subtype. [This invention 1136] The cell culture system of the present invention 1134, wherein the host cells are K562 cells containing exogenous HLA, the exogenous HLA is an HLA subtype defined by any one of HLA-peptide antigens in SEQ ID NOs: 10,755 to 29,364, and the cell culture medium contains a restriction peptide defined by the same HLA-peptide antigen that defines the HLA subtype. [This invention 1137] A method of identifying an antigen binding protein, comprising providing at least one HLA-peptide antigen as set forth in SEQ ID NOs: 10,755 to 29,364; and binding at least one target to any of the antigen binding proteins of the preceding invention, thereby identifying said antigen binding protein. [This invention 1138] 1137. The method of claim 1137, wherein said antigen binding protein is present in a phage display library comprising a plurality of distinct antigen binding proteins. [This invention 1139] 1138. The method of claim 1138, wherein said phage display library is substantially free of antigen-binding proteins that non-specifically bind to HLA of said HLA-peptide antigens. [The present invention 1140] 1139. The method of any of claims 1137 to 1139, wherein said combining step is performed two or more times, optionally at least three times. [This invention 1141] 1141. Any of the methods of inventions 1137 to 1140, further comprising contacting said antigen binding protein with one or more peptide-HLA complexes distinct from said HLA-peptide antigen to determine whether said antigen binding protein selectively binds to said HLA-peptide antigen, optionally wherein selectivity is determined by measuring the binding affinity of said antigen binding protein to a soluble target HLA-peptide complex and to a soluble HLA-peptide complex distinct from the target complex, and optionally wherein selectivity is determined by measuring the binding affinity of said antigen binding protein to a target HLA-peptide complex expressed on the surface of one or more cells and to an HLA-peptide complex distinct from the target complex expressed on the surface of one or more cells. [This invention 1142] 1. A method of identifying an antigen binding protein comprising obtaining at least one HLA-peptide antigen set forth in SEQ ID NOs: 10,755-29,364; administering said HLA-peptide antigen to a subject, optionally in combination with an adjuvant; and isolating any of the antigen binding proteins of the preceding invention from said subject. [This invention 1143] 1143. The method of claim 1142, wherein isolating said antigen binding protein comprises screening serum of said subject to identify said antigen binding protein. [This invention 1144] 1143. The method of claim 1142, further comprising contacting said antigen binding protein with one or more peptide-HLA complexes distinct from said HLA-peptide antigen to determine whether said antigen binding protein selectively binds to said HLA-peptide antigen, optionally wherein selectivity is determined by measuring the binding affinity of said antigen binding protein to said HLA-peptide antigen and to a soluble HLA-peptide complex distinct from said HLA-peptide antigen, optionally wherein selectivity is determined by measuring the binding affinity of said antigen binding protein to said HLA-peptide antigen expressed on the surface of one or more cells and to an HLA-peptide complex distinct from said HLA-peptide antigen expressed on the surface of one or more cells. [Invention 1145] 1142. The method of claim 1142, wherein said subject is a mouse, rabbit, or llama. [Invention 1146] 1143. The method of claim 1142, wherein isolating said antigen binding protein comprises isolating B cells from said subject which express said antigen binding protein, and optionally directly cloning a sequence encoding said antigen binding protein from said isolated B cells. [This invention 1147] The method of claim 1146, further comprising using said B cells to produce hybridomas. [This invention 1148] The method of claim 1146, further comprising cloning CDRs from said B cells. [This invention 1149] 1147. The method of claim 1146, further comprising immortalizing said B cells, optionally via EBV transformation. [This invention 1150] 1147. The method of claim 1146, further comprising generating a library comprising said antigen binding proteins of said B cells, optionally wherein said library is a phage display or yeast display. [This invention 1151] 1143. The method of claim 1142, further comprising humanizing said antigen-binding protein. [This invention 1152] A method for identifying an antigen-binding protein of any of the preceding inventions, comprising: obtaining a cell comprising the antigen-binding protein; contacting the cell with an HLA multimer comprising at least one HLA-peptide antigen set forth in SEQ ID NOs: 10,755 to 29,364; and identifying the antigen-binding protein via binding between the HLA multimer and the antigen-binding protein. [This invention 1153] 1152. The method of claim 1152, further comprising contacting said cell comprising said antigen binding protein with an HLA multimer comprising a corresponding wild-type sequence of said at least one HLA-peptide antigen as set forth in SEQ ID NOs: 10,755 to 29,364, and excluding said antigen binding protein if said antigen binding protein binds to said HLA multimer comprising said corresponding wild-type sequence. [This invention 1154] A method of identifying an antigen binding protein comprising providing at least one HLA-peptide antigen set forth in SEQ ID NOs: 10,755-29,364; and identifying said antigen binding protein of any of the preceding inventions using the target. [This invention 1155] 1. An antigen binding protein (ABP) that specifically binds to an HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimeric portion of the HLA class I molecule, wherein the HLA class I molecule and the HLA-restricted peptide are each selected from the HLA-peptide antigens set forth in any one of SEQ ID NOS: 10,755 to 29,364, and the ABP comprises an alpha CDR3 amino acid sequence and a corresponding beta CDR3 amino acid sequence selected from the group consisting of the sequences set forth in Tables 1C.1, 1C.2, 1C.3, and 1D. [Invention 1156] The ABP of the present invention 1155 further comprises an alpha variable ("V") segment, an alpha joining ("J") segment, a beta variable ("V") segment, a beta joining ("J") segment, optionally a beta diversity ("D") segment, and optionally a beta constant region selected from the group consisting of the regions set forth in Tables 1C.1, 1C.2, 1C.3, and 1D corresponding to said alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence. [This invention 1157] The ABP of any of 1155 or 1156 of the invention, comprising an alpha variable region and a corresponding beta variable region comprising an amino acid sequence selected from the sequences set forth in Tables 1A.1, 1A.2, 1A.3, and 1B corresponding to said alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence. [This invention 1158] 1. An antigen binding protein (ABP) that specifically binds to an HLA-peptide antigen comprising an HLA-restricted RAS peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, the HLA-restricted RAS peptide comprises at least one alteration that renders the HLA-restricted RAS peptide sequence different from the corresponding peptide sequence of a wild-type RAS peptide, and the ABP comprises an alpha CDR3 amino acid sequence and a corresponding beta CDR3 amino acid sequence selected from the group consisting of the sequences shown in Tables 1C.1, 1C.2, 1C.3, and 1D. [This invention 1159] a. the restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-A*02:01; b. the restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-A*02:06; c. The restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*27:05; d. The restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*35:01; e. the restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*41:02; f. The restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*48:01; g. The restriction peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-C*08:03; h. The restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; i. the restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; j. The restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*03:02; k. The restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*68:01; l. The restriction peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-B*27:05; m. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*02:01; n. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*02:05; o. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*03:01; p. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*11:01; q. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*11:01; r. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*26:01; s. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*31:01; t. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*68:01; u. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*07:02; v. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*08:01; w. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*13:02; x. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*15:01; y. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*27:05; z. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*35:01; aa. the restriction peptide comprises the RAS_G12D mutation and the HLA class I molecule is HLA-B*37:01; bb. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*38:01; cc. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*40:01; dd. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*40:02; ee. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*44:02; ff. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*44:03; gg. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*48:01; hh. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*50:01; ii. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*57:01; jj. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*01:02; kk. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*02:02; ll. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*03:03; mm. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*03:04; nn. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*04:01; oo. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*05:01; pp. whether the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*07:04; qq. The restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*08:02; rr. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*08:03; ss. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*16:01; tt. the restriction peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*17:01; uu. The restriction peptide contains the RAS_G12R mutation and the HLA class I molecule is HLA-B*41:02; vv. the restriction peptide contains the RAS_G12R mutation and the HLA class I molecule is HLA-C*07:04; ww. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:01; xx. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:05; yy. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:06; zz. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*03:01; aaa. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*03:01; bbb. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*11:01; ccc. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*11:01; ddd. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*25:01; eee. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*26:01; fff. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*30:01; ggg. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*31:01; hhh. The restriction peptide contains the RAS_G12V mutation, and the HLA class I molecule is HLA-A*31:01; iii. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*32:01; jjj. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*68:02; kkk. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*07:02; III. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*08:01; mmm. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*13:02; nnn. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*14:02; ooo. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*15:01; ppp. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*27:05; qqq. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*39:01; rrr. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*40:01; sss. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*40:02; ttt. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*41:02; uuu. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*44:05; vvv. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*50:01; www. the restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*51:01; xxx. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*01:02; yyy. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*01:02; zzz. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*03:03; aaaa. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*03:04; bbbb. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*08:02; cccc. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*14:02; dddd. The restriction peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*17:01; eeee. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-A*02:01; ffff. the restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*07:02; gggg. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*08:01; hhhh. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:01; iiii. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:03; jjjj. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:08; kkkk. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*38:01; III. The restriction peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-C*04:01; mmmm. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*01:01; nnnn. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*02:01; oooo. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*23:01; pppp. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*29:01; qqqq. Whether the restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*30:02; rrrr. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*33:01; ssss. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*68:01; tttt. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*07:02; uuuu. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*08:01; vvvv. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*18:01; wwww. the restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*35:01; xxxx. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*38:01; yyyy. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*40:01; zzzz. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*44:02; aaaaa. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-C*03:04; bbbbb. The restriction peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-C*05:01; or ccccc. The restriction peptide contains the KRAS_Q61H mutation, and the HLA class I molecule is HLA-C*08:02. ABP of the present invention 1158. [The present invention 1160] 1158. The ABP of claim 1159, wherein said HLA-peptide antigen is a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV. [This invention 1161] An ABP of the present invention 1160 comprising an alpha CDR3 amino acid sequence and a corresponding beta CDR3 amino acid sequence selected from the group consisting of the sequences shown in Table 1C.2. [This invention 1162] The ABP of the present invention 1161 further comprising an alpha variable ("V") segment, an alpha joining ("J") segment, a beta variable ("V") segment, a beta joining ("J") segment, optionally a beta diversity ("D") segment, and optionally a beta constant region selected from the group consisting of the regions set forth in Table 1C.2 corresponding to said alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence. [This invention 1163] The ABP of the invention 1161 or 1162, comprising an alpha variable region and a corresponding beta variable region comprising an amino acid sequence selected from the sequences shown in Table 1A.2 corresponding to said alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence. [Invention 1164] 1158. The ABP of claim 1159, wherein said HLA-peptide antigen is a RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVGAVGVGK. [This invention 1165] An ABP of the present invention 1164 comprising an alpha CDR3 amino acid sequence and a corresponding beta CDR3 amino acid sequence selected from the group consisting of the sequences shown in Table 1C.3. [Invention 1166] The ABP of the present invention 1165 further comprising an alpha variable ("V") segment, an alpha joining ("J") segment, a beta variable ("V") segment, a beta joining ("J") segment, optionally a beta diversity ("D") segment, and optionally a beta constant region selected from the group consisting of the regions set forth in Table 1C.3 corresponding to said alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence. [This invention 1167] The ABP of the invention 1165 or 1166, comprising an alpha variable region and a corresponding beta variable region comprising an amino acid sequence selected from the sequences shown in Table 1A.3 corresponding to said alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence. [Brief explanation of the drawings]

[0086] These and other features, aspects and advantages of the present invention will become better understood with reference to the following description and accompanying drawings. [Figure 1] General structure of a human leukocyte antigen (HLA) class I molecule. By user atropos235 on en.wikipedia - Own work, CC BY 2.5, https: / / commons.wikimedia.org / w / index.php?curid=1805424. [Figure 2] Flow cytometry analysis of enriched naive and memory T cells is shown. Cells labeled with a pool of six neoantigen-MHC tetramers ("HLA / SNA") are shown to identify neoantigen-specific T cells (left panel, X-axis) and the corresponding wild-type peptide MHC-tetramer pool ("HLA / wildtype"; left panel, Y-axis). Cells labeled for the memory T cell phenotypic marker CD45RO are also shown (right panel). [Figure 3A]

[0023] Figure 1 shows flow cytometry analysis of expanded T cells previously sorted using a pool of six neoantigen-MHC tetramers ("HLA / SNA"). Shown are expanded cells labeled with each of the six neoantigen-MHC tetramers and their corresponding wild-type peptide-MHC tetramers. [Figure 3B]

[0023] Figure 1 shows flow cytometry analysis of expanded T cells previously sorted using neoantigen-MHC tetramers ("HLA / SNA"). Shown are expanded cells labeled with each of the four neoantigen-MHC tetramers and their corresponding wild-type peptide-MHC tetramers. [Figure 4] 1 shows the correlation between EDGE score and the probability of detecting a candidate shared neo-antigenic peptide by targeted mass spectrometry. [Figure 5] Figure 5A shows the flow cytometry gating strategy for detecting CD8+ T cells, and Figure 5B shows flow cytometry results demonstrating that the majority of CD8+ T cells exhibit binding to RAS G12V:HLA*1101 PHLA. [Figure 6] Flow cytometry analysis of expanded T cells previously sorted using a single neoantigen-MHC tetramer for two different donors. Shown are expanded cells labeled with each of the three neoantigen-MHC tetramers and their corresponding wild-type peptide-MHC tetramers. [Figure 7] 1 shows the titration of DOX administration in modulating the expression of representative neoantigens under the Tet-On system in multiple K562-HLA cell lines. [Figure 8] A representative KRAS G12V peptide, VVGAVGVGK, observed by mass spectrometry in the HLA-A*11:01 expressing K562 cell line is shown. The top panel shows that detection was dependent on DOX (left column no DOX, right panel with DOX added), and the bottom panel shows equivalent detection of the heavy peptide control. [Figure 9] Proliferating naive CD8 T cells gated on CD137+ after neoantigen (left panel) and DMSO (right panel) stimulation are shown. [Figure 10] A summary of in silico analysis of TCR sequences shared between (i) neoantigen-tetramer labeled cells; (ii) CD137+ neoantigen stimulated cells; and (iii) CD137+ DMSO stimulated cells is shown. [Figure 11A]Representative flow cytometry evaluation of TCR clone 01CA019_064_F05_0047 is shown. Activation markers CD25 (left panel), CD69 (middle panel), and CD137 (right panel) are shown for primary T cells transduced with the indicated TCRs and stimulated with the cognate neoantigen (lower panel) or the corresponding wild-type peptide (upper panel). [Figure 11B] Representative flow cytometry evaluation of TCR clone 01CA019_064_F05_0005 shows activation markers CD25 (left panel), CD69 (middle panel), and CD137 (right panel) of primary T cells transduced with the indicated TCRs and stimulated with the cognate neoantigen (lower panel) or the corresponding wild-type peptide (upper panel). [Figure 12] Figure 1 shows the proliferation of primary T cells transduced with the indicated candidate TCRs. The percentage of T cells containing diluted CellTraceViolet dye after co-culture with peptide-loaded APCs is shown. DETAILED DESCRIPTION OF THE INVENTION

[0087] Detailed Description Unless otherwise defined, all technical terms, notations, and other scientific terms used herein are intended to have the meaning commonly understood by those of ordinary skill in the art. In some cases, terms with commonly understood meanings are defined herein for clarity and / or ready reference, and the inclusion of such definitions herein should not necessarily be construed as representing a difference from that commonly understood in the art. The techniques and procedures described or referenced herein are generally well understood and routinely employed by those of ordinary skill in the art using conventional methodology, such as the widely used molecular cloning method described in Sambrook et al., Molecular Cloning: A LABORATORY MANUAL (4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY). Where appropriate, procedures involving the use of commercially available kits and reagents are generally performed according to manufacturer-defined protocols and conditions unless otherwise noted.

[0088] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Terms such as "including," "e.g.," and the like are intended to convey inclusion without limitation unless specifically stated otherwise.

[0089] As used herein, the term "comprising" specifically includes embodiments "consisting of" and "consisting essentially of" the listed elements, unless expressly stated otherwise.

[0090] The term "about" refers to and encompasses the indicated value and the range above and below that value. In certain embodiments, the term "about" refers to the specified value ±10%, ±5%, or ±1%. In certain embodiments, where applicable, the term "about" refers to the specified value(s) ±1 standard deviation of that value(s).

[0091] The term "antigen binding protein" or "ABP" is used herein in the broadest sense and includes a specific type of molecule that contains one or more antigen-binding domains that specifically bind to an antigen or epitope.

[0092] In some embodiments, the ABP comprises a TCR. In some embodiments, the ABP consists of a TCR. In some embodiments, the ABP consists essentially of a TCR. ABPs specifically include intact TCRs, TCR fragments, and ABP fragments. In some embodiments, the ABP comprises an alternative scaffold. In some embodiments, the ABP consists of an alternative scaffold. In some embodiments, the ABP consists essentially of an alternative scaffold. In some embodiments, the ABP comprises a TCR fragment. In some embodiments, the ABP consists of a TCR fragment. In some embodiments, the ABP consists essentially of a TCR fragment.

[0093] As provided herein, an "HLA-peptide ABP," "anti-HLA-peptide ABP," or "HLA-peptide-specific ABP" is an ABP that specifically binds to an antigen HLA-peptide. ABPs include proteins that contain one or more antigen-binding domains that specifically bind to an antigen or epitope via a variable region, such as a variable region derived from a T cell (e.g., a TCR).

[0094] As used herein, "variable region" refers to a variable nucleotide sequence that results from a recombination event; for example, a variable region may include the V, J, and / or D segments of a T cell receptor (TCR) sequence from a T cell, such as an activated T cell.

[0095] The term "antigen-binding domain" refers to a portion of an ABP that can specifically bind to an antigen or epitope. The antigen-binding domain may include the CDRs of a TCR, such as αCDR1, αCDR2, αCDR3, βCDR1, βCDR2, and βCDR3. The CDRs of a TCR are described herein.

[0096] The amino acid sequence boundaries of TCR CDRs can be determined by one of skill in the art using any of a number of known numbering schemes, including, but not limited to, the IMGT specific numbering scheme described in LeFranc, M.-P, Immunol Today. 1997 Nov;18(11):509; Lefranc, M.-P., "IMGT Locus on Focus: A new section of Experimental and Clinical Immunogenetics", Exp. Clin. Immunogenet., 15, 1-7 (1998); Lefranc and Lefranc, The T Cell Receptor FactsBook; and M.-P. Lefranc / Developmental and Comparative Immunology 27 (2003) 55-77, all of which are incorporated by reference.

[0097] An "ABP fragment" comprises a portion of an intact ABP, such as the antigen-binding or variable region of the intact ABP. ABP fragments include, for example, TCR fragments.

[0098] The term "alternative scaffold" refers to a molecule in which one or more regions can be diversified to generate one or more antigen-binding domains that specifically bind to an antigen or epitope. In some embodiments, the antigen-binding domain binds to an antigen or epitope with specificity and affinity comparable to that of an ABP. Exemplary alternative scaffolds include those derived from fibronectin (e.g., Adnectin™), β-sandwich (e.g., iMab), lipocalin (e.g., Anticalin®), EETI-II / AGRP, BPTI / LACI-D1 / ITI-D2 (e.g., Kunitz domain), thioredoxin peptide aptamer, protein A (e.g., Affibody®), ankyrin repeat (e.g., DARPin), gamma-B-crystallin / ubiquitin (e.g., Affilin), CTLD3 (e.g., tetranectin), Fynomer, and (LDLR-A module) (e.g., Avimer). Further information on alternative scaffolds is provided in Binz et al., Nat. Biotechnol., 2005 23:1257-1268; Skerra, Current Opin. in Biotech., 2007 18:295-304; and Silacci et al., J. Biol. Chem., 2014, 289:14392-14398, each of which is incorporated by reference in its entirety. Alternative scaffolds are a type of ABP.

[0099] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an ABP) and its binding partner (e.g., an antigen or epitope). Unless otherwise specified, as used herein, "affinity" refers to the intrinsic binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., an ABP and an antigen or epitope). The affinity of a molecule X for its partner Y is determined by the dissociation equilibrium constant (K D) The kinetic components that contribute to the dissociation equilibrium constant are described in detail below. Affinity can be measured by common methods known in the art, including those described herein, such as surface plasmon resonance (SPR) technology (e.g., BIACORE®) or biolayer interferometry (e.g., FORTEBIO®).

[0100] The terms "bind," "specific binding," "specifically binds," "specific," "selectively binds," and "selective" refer to binding of an ABP to a target molecule that is measurably different from nonspecific or nonselective interactions (e.g., interactions with non-target molecules) of a particular antigen (e.g., a polypeptide target) or epitope on a particular antigen. Specific binding can be measured, for example, by measuring binding to a target molecule and comparing it to binding to a non-target molecule. Specific binding can also be determined by competition with a control molecule that mimics the epitope recognized by the target molecule. Specific binding is indicated when binding of the ABP to the target molecule is competitively inhibited by the control molecule. In some embodiments, the affinity of the HLA peptide ABP for the non-target molecule is less than about 50% of its affinity for the HLA-peptide. In some embodiments, the affinity of the HLA peptide ABP for the non-target molecule is less than about 40% of its affinity for the HLA-peptide. In some embodiments, the affinity of the HLA peptide ABP for the non-target molecule is less than about 30% of its affinity for the HLA-peptide. In some embodiments, the affinity of the HLA peptide ABP for the non-target molecule is less than about 20% of its affinity for the HLA-peptide. In some embodiments, the affinity of the HLA peptide ABP for the non-target molecule is less than about 10% of its affinity for the HLA-peptide. In some embodiments, the affinity of the HLA peptide ABP for the non-target molecule is less than about 1% of its affinity for the HLA-peptide. In some embodiments, the affinity of the HLA peptide ABP for the non-target molecule is less than about 0.1% of its affinity for the HLA-peptide.

[0101] As used herein, "k d ”(seconds -1 The term k ) refers to the dissociation rate constant of a particular ABP-antigen interaction. This value is off Also called value.

[0102] As used herein, "k a " (M -1 × seconds -1 The term k ) refers to the association rate constant for a particular ABP-antigen interaction. This value is on Also called value.

[0103] As used herein, "K D The term "(M)" refers to the dissociation equilibrium constant of a particular ABP-antigen interaction. D =k d / k a In some embodiments, the affinity of an ABP is determined by the K D For clarity, as known in the art, K D A smaller value indicates a higher affinity interaction, and K D Higher values ​​indicate lower affinity interactions.

[0104] As used herein, "K A " (M -1 The term K ) refers to the association equilibrium constant of a particular ABP-antigen interaction. A =k a / k d .

[0105] An "immunoconjugate" is an ABP conjugated to one or more heterologous molecule(s), eg, a therapeutic agent (eg, a cytokine) or a diagnostic agent.

[0106] The terms "compete with" or "cross-compete with," when used herein in the context of two or more ABPs, indicate that the two or more ABPs compete for binding to an antigen (e.g., an HLA-peptide). In one exemplary assay, an HLA-peptide is coated on a surface and contacted with a first HLA-peptide ABP, followed by the addition of a second HLA-peptide ABP. In another exemplary assay, a first HLA-peptide ABP is coated on a surface and contacted with an HLA-peptide, followed by the addition of a second HLA-peptide ABP. In either assay, if the presence of the first HLA-peptide ABP reduces the binding of the second HLA-peptide ABP, then the ABPs compete with each other. The term "compete with" also includes combinations of ABPs in which one ABP reduces the binding of another ABP, but where no competition is observed when the ABPs are added in the reverse order. However, in some embodiments, the first and second ABPs inhibit each other's binding, regardless of the order in which they are added. In some embodiments, one ABP reduces the binding of another ABP to its antigen by at least 25%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, or at least 95%. One skilled in the art can select the concentration of ABP used in a competition assay based on the affinity of the ABP for the HLA-peptide and the valency of the ABP. The assays described in this definition are exemplary, and one skilled in the art can use any suitable assay to determine whether ABPs compete with each other.Suitable assays are described, for example, in Cox et al., "Immunoassay Methods," in Assay Guidance Manual [Internet], Updated December 24, 2014 (www.ncbi.nlm.nih.gov / books / NBK92434 / ; accessed September 29, 2015); Silman et al., Cytometry, 2001, 44:30-37; and Finco et al., J. Pharm. Biomed. Anal., 2011, 54:351-358, which are incorporated by reference in their entireties.

[0107] The term "epitope" refers to the portion of an antigen that specifically binds to an ABP. Epitopes often consist of surface-accessible amino acid residues and / or sugar side chains and may have specific three-dimensional structural and charge characteristics. Conformational and nonconformational epitopes are distinguished in that binding to the former, but not the latter, can be lost in the presence of denaturing solvents. An epitope may include amino acid residues directly involved in binding and other amino acid residues not directly involved in binding. The epitope to which an ABP binds can be determined using known techniques for epitope determination, such as testing ABP binding to HLA-peptide variants with different point mutations or chimeric HLA-peptide variants.

[0108] As used herein, the term "percent identity," in the context of two or more nucleic acid or polypeptide sequences, refers to two or more sequences or subsequences that have a specified percentage of nucleotides or amino acid residues that are identical when compared and aligned for maximum correspondence, as measured using one of the sequence comparison algorithms described below (e.g., BLASTP and BLASTN or other algorithms available to those of skill in the art) or by visual inspection. Depending on the application, the percent "identity" may exist over a region of the sequences being compared, e.g., over a functional domain, or over the entire length of the two sequences being compared.

[0109] For sequence comparison, typically, one sequence serves as a reference sequence to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are input into a computer, subsequence coordinates are designated as necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity of the test sequence(s) relative to the reference sequence based on the designated program parameters. Alternatively, sequence similarity or dissimilarity can be established by the presence or absence of specific nucleotides, or in the case of translated sequences, amino acid combinations at selected sequence positions (e.g., sequence motifs).

[0110] Optimal alignment of sequences for comparison may be achieved, for example, by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA, in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection (see generally Ausubel et al., infra).

[0111] One example of an algorithm that is suitable for determining percent sequence identity and sequence similarity is the BLAST algorithm described in Altschul et al., J. Mol. Biol. 215:403-410 (1990). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information.

[0112] "Conservative substitution" or "conservative amino acid substitution" refers to the substitution of an amino acid with a chemically or functionally similar amino acid. Conservative substitution tables providing similar amino acids are well known in the art. As examples, the groups of amino acids provided in Tables 2-4 are, in some embodiments, considered conservative substitutions for each other.

[0113] Table 2: Selected groups of amino acids that are considered conservative substitutions for one another in certain embodiments. TIFF2025176716000002.tif32128

[0114] Table 3: Additional selected groups of amino acids that are considered conservative substitutions for one another in certain embodiments. TIFF2025176716000003.tif32128

[0115] Table 4: Further selected groups of amino acids that are considered conservative substitutions for one another in certain embodiments. TIFF2025176716000004.tif43128

[0116] Additional conservative substitutions can be found, for example, in Creighton, Proteins: Structures and Molecular Properties 2nd ed. (1993) W.H. Freeman & Co., New York, NY. ABPs generated by making one or more conservative substitutions of amino acid residues of a parent ABP are referred to as "conservatively modified variants."

[0117] The term "amino acid" refers to the 20 common naturally occurring amino acids, including alanine (Ala; A), arginine (Arg; R), asparagine (Asn; N), aspartic acid (Asp; D), cysteine ​​(Cys; C), glutamic acid (Glu; E), glutamine (Gln; Q), glycine (Gly; G), histidine (His; H), isoleucine (Ile; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P), serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr; Y), and valine (Val; V).

[0118] As used herein, the term "vector" refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes vectors as self-replicating nucleic acid structures as well as vectors that integrate into the genome of a host cell into which they are introduced. Certain vectors are capable of driving the expression of nucleic acids to which they are operatively linked. Such vectors are referred to herein as "expression vectors."

[0119] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, and the progeny of such cells. Host cells include "transformants" (or "transformed cells") and "transformants" (or "transfected cells"), which include the primary transformed or transfected cell, respectively, and their derived progeny. Such progeny may not necessarily be completely identical in nucleic acid content to the parent cell and may contain mutations.

[0120] The term "treat" (and variations thereof, such as "treating" or "treatment") refers to clinical intervention in an attempt to alter the natural course of a disease or condition in a subject in need thereof. Treatment can be performed both prophylactically and during the course of clinical pathology. Desirable effects of treatment include prevention of disease onset or recurrence, alleviation of symptoms, reduction of the direct or indirect pathological effects of disease, prevention of metastasis, slowing the rate of disease progression, improvement or palliation of the disease state, and remission or improved prognosis.

[0121] As used herein, the term "therapeutically effective amount" or "effective amount" refers to the amount of an ABP or pharmaceutical composition provided herein that, when administered to a subject, is effective to treat a disease or disorder.

[0122] As used herein, the term "subject" refers to a mammalian subject. Exemplary subjects include humans, monkeys, dogs, cats, mice, rats, cows, horses, camels, goats, rabbits, and sheep. In certain embodiments, the subject is a human. In some embodiments, the subject's disease or condition can be treated with the ABPs provided herein. In some aspects, the disease or condition is cancer. In some aspects, the disease or condition is a viral infection.

[0123] The term "package insert" is used to refer to the instructions normally included in the commercial packaging of a therapeutic or diagnostic product, which contain information about the indications, uses, dosage, administration, combination therapy, contraindications and / or warnings regarding the use of such therapeutic or diagnostic product.

[0124] The term "tumor" refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all pre-cancerous and cancerous cells and tissues. The terms "cancer," "cancerous," "cell proliferative disorder," "proliferative disorder," and "tumor" are not mutually exclusive when referred to herein. The terms "cell proliferative disorder" and "proliferative disorder" refer to disorders associated with some degree of abnormal cell proliferation. In some embodiments, the cell proliferative disorder is cancer. In some aspects, the tumor is a solid tumor. In some aspects, the tumor is a hematological malignancy.

[0125] The term "pharmaceutical composition" refers to a preparation that is in a form that allows the biological activity of the active ingredient contained therein to be effective in treating a subject, and that does not contain additional ingredients that are unacceptably toxic to a subject in the amounts provided in the pharmaceutical composition.

[0126] The terms "modulate" and "modulation" refer to decreasing or inhibiting, or alternatively activating or increasing, the recited variable.

[0127] The terms "increase" and "activate" refer to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold or greater increment in the recited variable.

[0128] The terms "reduce" and "inhibit" refer to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold or greater decrement in the recited variable.

[0129] The term "stimulate" refers to activating receptor signaling to induce a biological response associated with receptor activation. An "agonist" is an entity that binds to and stimulates a receptor.

[0130] The term "antagonize" refers to inhibiting a biological response associated with receptor activation by inhibiting receptor signaling. An "antagonist" is an entity that binds to and antagonizes a receptor.

[0131] The terms "nucleic acid" and "polynucleotide" may be used interchangeably herein and refer to a polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or their analogs. Polynucleotides can include, but are not limited to, coding or non-coding regions of a gene or gene fragment, loci (gene loci) revealed by linkage analysis, exons, introns, messenger RNA (mRNA), cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA, isolated RNA, nucleic acid probes, and primers. Polynucleotides can contain modified nucleotides, such as methylated nucleotides and nucleotide analogs. Exemplary modified nucleotides include, for example, 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxymethyl)uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, β-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-substituted adenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, and N6-substituted adenine. Examples include denine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, β-D-mannosylqueosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthioN6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxocine, pseudouracil, quosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methyl ester, 3-(3-amino-3-N-2-carboxypropyl)uracil, and 2,6-diaminopurine.

[0132] As used herein, the term "antigen" refers to a substance that induces an immune response. The antigen may be a neoantigen. The antigen may also be a "shared antigen," which is an antigen found among a particular population, for example, a particular population of cancer patients. Antigens include HLA-peptide antigens.

[0133] As used herein, the term "neoantigen" refers to an antigen that has at least one alteration that distinguishes it from the corresponding wild-type antigen, for example, through a mutation in tumor cells or a tumor cell-specific post-translational modification. In some embodiments, the alteration occurs in tumor or cancer cells. In some embodiments, the alteration does not occur in non-tumor or non-cancer cells. In some embodiments, the alteration is not present in normal tissues. Neoantigens can include polypeptide or nucleotide sequences. Mutations can include frameshift or non-frameshift indels, missense or nonsense substitutions, splice site alterations, genomic rearrangements or gene fusions, or any genomic or expression alterations that result in neo-ORFs. Mutations can also include splice variants. Tumor cell-specific post-translational modifications can include aberrant phosphorylation. Tumor cell-specific post-translational modifications can also include spliced ​​antigens generated by the proteasome. See Liepe et al., A large fraction of HLA class I ligands are proteasome-generated spliced ​​peptides; Science. 2016 Oct 21;354(6310):354-358. Neoantigens may be shared neoantigens if they are found among multiple patients in a particular population (e.g., a particular population of cancer patients). Neoantigens may also include HLA-peptide neoantigens.

[0134] As used herein, the terms "HLA-peptide," "pHLA," "peptide-HLA," and "peptide-HLA complex" are used interchangeably herein to refer to an antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, where the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule. Such antigens are defined by a specific HLA-restricted peptide having a defined amino acid sequence complexed with a specific HLA class I subtype.

[0135] In some embodiments, "HLA-peptide neoantigen," "pHLA neoantigen," and "peptide-HLA neoantigen" are used interchangeably herein to refer to an HLA-peptide that contains at least one alteration that distinguishes it from the corresponding wild-type HLA-peptide antigen, e.g., via a mutation in a tumor cell or a tumor cell-specific post-translational modification. In some embodiments, the at least one alteration is in the constrained peptide sequence, such that the constrained peptide of the HLA-peptide neoantigen is distinguishable from a constrained peptide that contains the corresponding constrained peptide sequence without the alteration, e.g., the wild-type sequence.

[0136] Exemplary HLA-peptide neoantigens and shared HLA-peptide neoantigens are shown in Table A (SEQ ID NOS: 10,755-21,015), AACR GENIE results (SEQ ID NOS: 21,016-29,357), and SEQ ID NOS: 29358-29364; the corresponding genes and somatic alterations associated with each antigen are also shown. Such pHLA neoantigens and shared pHLA neoantigens are useful for inducing an immune response in a subject via administration. Subjects for administration may be identified using various diagnostic methods, such as the patient selection methods described herein.

[0137] As used herein, the term "tumor antigen" is an antigen derived from a polypeptide that is known or found to be present in tumor cells or tissues of a subject but not in the subject's corresponding normal cells or tissues, or whose expression is altered in tumor cells or cancerous tissues compared to normal cells or tissues.

[0138] As used herein, the term "candidate antigen" is a mutation or other abnormality that gives rise to a sequence that may represent an antigen.

[0139] As used herein, the term "coding region" is the portion or portions of a gene that encode a protein.

[0140] As used herein, the term "coding mutation" is a mutation that occurs in a coding region.

[0141] As used herein, the term "ORF" means open reading frame.

[0142] As used herein, the term "neo-ORF" is a tumor-specific ORF that results from mutation or other deviations such as splicing.

[0143] As used herein, the term "missense mutation" is a mutation that results in the substitution of one amino acid for another.

[0144] As used herein, the term "nonsense mutation" is a mutation that results in the substitution of an amino acid with a stop codon or the removal of the canonical start codon.

[0145] As used herein, the term "frameshift mutation" is a mutation that causes an alteration in the frame of a protein.

[0146] As used herein, the term "indel" is an insertion or deletion of one or more nucleic acids.

[0147] As used herein, the term "non-stop or read-through" refers to a mutation that results in the removal of the natural stop codon.

[0148] HLA-peptide antigen The major histocompatibility complex (MHC) is a complex encoded by a group of linked genetic loci, collectively referred to as H-2 in mice and HLA in humans. The two major classes of MHC antigens, class I and class II, each contain a series of cell surface glycoproteins that play a role in determining tissue type and transplant compatibility. In the transplant response, cytotoxic T cells (CTLs) respond primarily to class I glycoproteins, whereas helper T cells respond primarily to class II glycoproteins.

[0149] Human major histocompatibility complex (MHC) class I molecules, referred to interchangeably herein as HLA class I molecules, are expressed on the surface of nearly all cells. These molecules function to present peptides, most of which are derived from endogenously synthesized proteins, to, for example, CD8+ T cells via interaction with the α-β T cell receptor. Class I MHC molecules contain heterodimers composed of a 46 kDa α chain noncovalently associated with a 12 kDa light chain β2-microglobulin. The α chain generally contains α1 and α2 domains, which form a groove for presenting HLA-restricted peptides and an α3 transmembrane domain that interacts with the CD8 coreceptor on T cells. Figure 1 depicts the general structure of a class I HLA molecule. Some TCRs can bind to MHC class I independently of the CD8 co-receptor (see, e.g., Kerry SE, Buslepp J, Cramer LA, et al. Interplay between TCR Affinity and Necessity of Coreceptor Ligation: High-Affinity Peptide-MHC / TCR Interaction Overcomes Lack of CD8 Engagement. Journal of immunology (Baltimore, Md: 1950). 2003;171(9):4493-4503).

[0150] Class I MHC-restricted peptides (also referred to herein interchangeably as HLA-restricted antigens, HLA-restricted peptides, antigenic peptides, MHC-restricted antigens, restricted peptides, or peptides) typically bind to the α1-α2 groove of the heavy chain via approximately two or three anchor residues that interact with the corresponding binding pocket of the MHC molecule. The β-2 microglobulin chain plays a critical role in intracellular trafficking, peptide binding, and structural stability of MHC class I. For most class I molecules, the formation of a heterotrimeric complex of the MHC class I heavy chain, peptide (self, non-self, and / or antigenic), and β2 microglobulin results in protein maturation and transport to the cell surface.

[0151] Binding of a given HLA subtype to an HLA-restricted peptide results in the formation of a complex with a unique and novel surface that can be specifically recognized by an ABP, for example, a TCR on a T cell.

[0152] Thus, provided herein are HLA-peptide antigens that include specific HLA-restricted peptides having defined amino acid sequences that complex with specific HLA subtypes.

[0153] The HLA-peptide antigens identified herein can be useful for cancer immunotherapy. In some embodiments, the HLA-peptide targets identified herein are presented on the surface of tumor cells. The HLA-peptide antigens identified herein can be expressed by tumor cells of human subjects. The HLA-peptide antigens identified herein can be expressed by tumor cells in a population of human subjects. For example, the HLA-peptide antigens identified herein can be shared HLA-peptide antigens that are commonly expressed in a population of human subjects with cancer.

[0154] The HLA-peptide antigens identified herein may have a prevalence of individual tumor types, which may be approximately 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, It can be 5%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. The prevalence of individual tumor types can be approximately 0.1% to 100%, 0.2 to 50%, 0.5 to 25%, or 1 to 10%.

[0155] Exemplary HLA class I subtypes of pHLA neoantigens In humans, many MHC haplotypes exist (interchangeably referred to herein as MHC subtypes, HLA subtypes, MHC types, and HLA types). Exemplary HLA subtypes include, by way of example only, 2-digit, 4-digit, 6-digit, and 8-digit subtypes. A complete list of HLA class alleles can be found at http: / / hla.alleles.org / alleles / . For example, a complete list of HLA class I alleles can be found at http: / / hla.alleles.org / alleles / class1.html. Exemplary HLA class I subtypes include any of the HLA subtypes disclosed in Table A (see SEQ ID NOS: 10,755-21,015), in the AACR GENIE results (see SEQ ID NOS: 21,016-29,357), and in SEQ ID NOS: 29358-29364 disclosed herein. The neoantigens in Table A and the AACR GENIE results are disclosed in PCT / US2019 / 033830, filed May 23, 2019, which is incorporated herein by reference in its entirety.

[0156] Exemplary HLA-Restricted Peptides An HLA-restricted peptide (interchangeably referred to herein as a "restricted peptide") can be a peptide fragment of a tumor-associated neoantigen, e.g., a shared neoantigen. The peptide fragment can include any of the amino acid sequences disclosed in Table A (see SEQ ID NOS: 10,755-21,015), in the AACR GENIE results (see SEQ ID NOS: 21,016-29,357), and in SEQ ID NOS: 29358-29364 disclosed herein. The neoantigens of Table A and the AACR GENIE results are disclosed in PCT / US2019 / 033830, filed May 23, 2019, which application is incorporated herein by reference in its entirety.

[0157] Thus, disclosed herein are isolated peptides containing tumor-specific mutations identified by the methods disclosed herein, peptides containing known tumor-specific mutations, and mutant polypeptides or fragments thereof identified by the methods disclosed herein. Neoantigen peptides can be described in the context of their coding sequences, and neoantigens include nucleotide sequences (e.g., DNA or RNA) that encode the relevant polypeptide sequence.

[0158] Also disclosed herein are peptides, such as restriction peptides derived from any polypeptide known or found to have altered expression in tumor cells or cancerous tissues compared to normal cells or tissues, for example, any polypeptide known or found to be abnormally expressed in tumor cells or cancerous tissues compared to normal cells or tissues. Suitable polypeptides from which restriction peptides can be derived can be found, for example, in the COSMIC database. COSMIC organizes comprehensive information on somatic mutations in human cancers. In some embodiments, the restriction peptide comprises a tumor-specific mutation.

[0159] The one or more constrained peptides may include at least one of the following: a binding affinity to MHC with an IC50 value of less than 1000 nM for MHC class I peptides 8-15, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids in length, the presence of a sequence motif within or near the peptide that promotes proteasomal cleavage, and the presence of a sequence motif that promotes TAP transport.

[0160] The constrained peptides may have a size of about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, or about 15 amino acid residues, or any range derivable therein. In certain embodiments, the constrained peptides have a size of about 8, about 9, about 10, about 11, or about 12 amino acid molecule residues. The constrained peptides may be about 5-15 amino acids in length, preferably about 7-13 amino acids in length, or more preferably about 8-12 amino acids in length.

[0161] Exemplary Shared HLA-Peptide Neoantigens Exemplary shared HLA-peptide neoantigens are shown in Table A (see SEQ ID NOS: 10,755-21,015), the AACR GENIE results (see SEQ ID NOS: 21,016-29,357), and SEQ ID NOS: 29358-29364 disclosed herein. The neoantigens in Table A and the AACR GENIE results are disclosed in PCT / US2019 / 033830, filed May 23, 2019, which application is incorporated herein by reference in its entirety.

[0162] One or more HLA-peptide neoantigens may be presented on the surface of the tumor.

[0163] One or more HLA-peptide neoantigens may be immunogenic in a tumor-bearing subject, for example, capable of eliciting a T-cell or B-cell response in the subject.

[0164] If necessary, longer peptides can be designed in several ways. In some cases, when the potential for peptide presentation on HLA alleles is predicted or known, longer peptides can consist of either: (1) individual presented peptides with 2–5 amino acid extensions to the N- and C-termini of the corresponding gene product; or (2) concatenation of part or all of the presented peptide with the respective extension sequence. In other cases, when sequencing reveals the presence of long (>10 residues) neoepitope sequences present in the tumor (e.g., due to frameshifts, readthrough, or intron inclusion leading to new peptide sequences), longer peptides can consist of: 3) entire stretches of novel tumor-specific amino acids (thereby avoiding the need for computational or in vitro test-based selection of shorter peptides for strongest HLA presentation). In both cases, using longer peptides allows for endogenous processing by patient cells, potentially leading to more effective antigen presentation and induction of T cell responses.

[0165] Antigenic peptides and polypeptides can be presented on HLA proteins. In some embodiments, antigenic peptides and polypeptides are presented on HLA proteins with higher affinity than wild-type peptides. In some embodiments, the antigenic peptide or polypeptide can have an IC50 of at least 5000 nM or less, at least 1000 nM or less, at least 500 nM or less, at least 250 nM or less, at least 200 nM or less, at least 150 nM or less, at least 100 nM or less, or at least 50 nM or less.

[0166] In some embodiments, the antigenic peptides and polypeptides do not induce an autoimmune response and / or do not induce immune tolerance when administered to a subject.

[0167] Compositions containing at least two or more antigenic peptides are also provided. In some embodiments, the composition comprises at least two distinct peptides. The at least two distinct peptides may be derived from the same polypeptide. Distinct polypeptides mean that the peptides vary in length, amino acid sequence, or both. The peptides may be derived from any polypeptide known or found to contain tumor-specific mutations, or any polypeptide known or found to have altered expression in tumor cells or cancerous tissues compared to normal cells or tissues, e.g., any polypeptide known or found to be aberrantly expressed in tumor cells or cancerous tissues compared to normal cells or tissues. Suitable polypeptides from which antigenic peptides can be derived can be found, for example, in the COSMIC database or the AACR Genomics Evidence Neoplasia Information Exchange (GENIE) database. COSMIC organizes comprehensive information on somatic mutations in human cancers. AACR GENIE aggregates clinical-grade cancer genomic data and associates it with clinical outcomes from tens of thousands of cancer patients. The peptides include tumor-specific mutations. In some embodiments, the tumor-specific mutation is a driver mutation of a particular cancer type.

[0168] Antigenic peptides and polypeptides with desired activities or properties can be modified to provide certain desired attributes, such as improved pharmacological properties, while increasing or at least retaining substantially all of the biological activity of the unmodified peptide for binding to desired MHC molecules and activating appropriate T cells. For example, antigenic peptides and polypeptides can undergo various changes, such as conservative or non-conservative substitutions, which may provide certain advantages in their use, such as improved MHC binding, stability, or presentation. Conservative substitutions refer to the replacement of an amino acid residue with another that is biologically and / or chemically similar, for example, a hydrophobic residue for another, or a polar residue for another. Substitutions include combinations such as Gly, Ala; Val, Ile, Leu, Met; Asp, Glu; Asn, Gln; Ser, Thr; Lys, Arg; and Phe, Tyr. The effects of single amino acid substitutions can also be examined using D-amino acids. Such modifications may be carried out using well-known peptide synthesis procedures, as described, for example, in Merrifield, Science 232:341-347 (1986), Barany & Merrifield, The Peptides, Gross & Meienhofer, eds. (NY, Academic Press), pp. 1-284 (1979); and Stewart & Young, Solid Phase Peptide Synthesis, (Rockford, Ill., Pierce), 2nd Ed. (1984).

[0169] Modification of peptides and polypeptides with various amino acid mimetics or unnatural amino acids can be particularly useful for increasing the stability of peptides and polypeptides in vivo. Stability can be analyzed in a number of ways. For example, peptidases and various biological media, such as human plasma and serum, have been used to test stability. See, for example, Verhoef et al., Eur. J. Drug Metab Pharmacokin. 11:291-302 (1986). Peptide half-life can be conveniently determined using a 25% human serum (v / v) assay. The protocol generally follows: Pooled human serum (type AB, non-heat-inactivated) is defatted by centrifugation before use. The serum is then diluted to 25% with RPMI tissue culture medium and used to test peptide stability. At predetermined time intervals, a small amount of the reaction solution is removed and added to either 6% aqueous trichloroacetic acid or ethanol. The cloudy reaction sample is cooled (4°C) for 15 minutes and then spun to pellet precipitated serum proteins. The presence of the peptide is then confirmed by reverse-phase HPLC using stability-specific chromatographic conditions.

[0170] Peptides and polypeptides can be modified to provide desired attributes other than improved serum half-life. For example, the ability of a peptide to induce CTL activity can be enhanced by linking it to a sequence containing at least one epitope capable of inducing a T helper cell response. The immunogenic peptide / T helper conjugate can be linked by a spacer molecule. The spacer is usually composed of relatively small, neutral molecules, such as amino acids or amino acid mimetics, that are substantially uncharged under physiological conditions. Spacers are typically selected from, for example, Ala, Gly, or other neutral spacers of nonpolar or neutral polar amino acids. It will be understood that the optional spacer need not be composed of the same residues and can therefore be a hetero- or homo-oligomer. If present, the spacer will typically be at least one or two residues, more usually three to six residues. Alternatively, the peptide can be linked to the T helper peptide without a spacer.

[0171] The antigenic peptide can be linked to a T helper peptide either directly or via a spacer at either the amino or carboxy terminus of the peptide. The amino terminus of either the antigenic peptide or the T helper peptide can be acylated. Exemplary T helper peptides include tetanus toxoid 830-843, influenza 307-319, malaria circumsporozoite 382-398, and 378-389.

[0172] Proteins or peptides can be produced by any technique known to those of skill in the art, including expressing the protein, polypeptide, or peptide by standard molecular biology techniques, isolating the protein or peptide from a natural source, or chemically synthesizing the protein or peptide. Nucleotide and protein, polypeptide, and peptide sequences corresponding to various genes have been previously disclosed and can be found in computerized databases known to those of skill in the art. One such database is the Genbank and GenPept databases of the National Center for Biotechnology Information at the National Institutes of Health website. The coding regions of known genes can be amplified and / or expressed using the techniques disclosed herein or as known to those of skill in the art. Alternatively, various commercially available preparations of proteins, polypeptides, and peptides are known to those of skill in the art.

[0173] In some embodiments, an antigen may comprise a nucleic acid (e.g., a polynucleotide) encoding an antigenic peptide or a portion thereof, which may be, for example, DNA, cDNA, PNA, CNA, RNA (e.g., mRNA), either single-stranded and / or double-stranded, or a naturally occurring or stabilized form of a polynucleotide, e.g., a polynucleotide having a phosphorothioate backbone, or a combination thereof, and may or may not contain introns.

[0174] A further embodiment provides an expression vector capable of expressing a polypeptide or a portion thereof. Expression vectors for different cell types are well known in the art and can be selected without undue experimentation. Generally, DNA is inserted into an expression vector, such as a plasmid, in the appropriate orientation and in the correct reading frame for expression. If necessary, the DNA can be linked to appropriate transcriptional and translational regulatory control nucleotide sequences recognized by the intended host; such controls are generally available in expression vectors. The vector is then introduced into the host using standard techniques. Guidance can be found, for example, in Sambrook et al. (1989) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

[0175] HLA class I molecules that do not associate with a tethered peptide ligand are generally unstable. Therefore, the association of a tethered peptide with the α1 / α2 groove of an HLA molecule may stabilize the noncovalent association of the β2-microglobulin subunit of an HLA subtype with the α subunit of an HLA subtype.

[0176] The stability of the non-covalent association between the β2 microglobulin subunit of an HLA subtype and the α subunit of an HLA subtype can be determined by any suitable means.For example, such stability can be evaluated by dissolving insoluble aggregates of HLA molecules in a high concentration of urea (for example, about 8M urea), and determining the ability of the HLA molecules to refold in the presence of a constrained peptide during urea removal, for example, by dialysis.Such refolding approaches are described, for example, in Proc.Natl.Acad.Sci.USA Vol.89, pp.3429-3433, April 1992, the entire contents of which are incorporated by reference.

[0177] In another example, such stability can be assessed using conditional HLA class I ligands. Conditional HLA class I ligands are generally designed as short constrained peptides that bind to the α1 / α2 groove of HLA molecules, thereby stabilizing the association of the β2 and α subunits of HLA class I molecules, and contain one or more amino acid modifications that allow the constrained peptide to cleave upon exposure to a conditional stimulus. Upon cleavage of the conditional ligand, the β2 and α subunits of the HLA molecule dissociate unless the conditional ligand is replaced with a constrained peptide that binds to the α1 / α2 groove and stabilizes the HLA molecule. Conditional ligands can be designed by introducing amino acid modifications into known or predicted high-affinity HLA-peptide ligands. Furthermore, for HLA alleles for which structural information is available, side chain water accessibility can be exploited to select positions for introducing amino acid modifications. The use of conditional HLA ligands can be advantageous because it allows batch preparation of stable HLA-peptide complexes, which can then be used to investigate test constrained peptides in a high-throughput manner.Conditional HLA class I ligands and methods for producing them are described, for example, in Proc Natl Acad Sci US A. 2008 Mar 11;105(10): 3831-3836; Proc Natl Acad Sci US A. 2008 Mar 11;105(10): 3825-3830; J Exp Med. 2018 May 7;215(5): 1493-1504; Choo, JAL et al. Bioorthogonal cleavage and exchange of major histocompatibility complex ligands by employing azobenzene-containing peptides. Angew Chem Int Ed Engl 53, 13390-13394 (2014); Amore, A. et al. Development of a Hypersensitive Periodate-Cleavable Amino Acid that is Methionine- and Disulfide-Compatible and Its Application in MHC Exchange. Reagents for T Cell Characterization. ChemBioChem 14, 123-131 (2012); Rodenko, B. et al. Class I Major Histocompatibility Complexes Loaded by a Periodate Trigger. J Am Chem Soc 131, 12305-12313 (2009); and Chang, CXL et al. Conditional ligands for Asian HLA variants facilitate the definition of CD8+ T-cell responses in acute and chronic viral diseases. Eur J Immunol 43, 1109-1120 (2013). These references are incorporated by reference in their entireties.

[0178] Thus, in some embodiments, the ability of HLA-restricted peptides described herein, e.g., in Table A (SEQ ID NOS: 10,755-21,015), in the AACR GENIE results (SEQ ID NOS: 21,016-29,357), or in SEQ ID NOS: 29358-29364, to stabilize the association of the β2 and α subunits of HLA molecules is assessed by performing a conditional ligand-mediated exchange reaction and an HLA stability assay. HLA stability can be assayed using any suitable method, including, for example, mass spectrometry, immunoassays (e.g., ELISA), size exclusion chromatography, HLA multimer staining followed by flow cytometric evaluation of T cells, etc.

[0179] Other exemplary methods for evaluating the stability of the non-covalent association between the β2 microglobulin subunit of HLA subtype and the α subunit of HLA subtype include peptide exchange using dipeptide.Peptide exchange using dipeptide is described in, for example, Proc Natl Acad Sci US A. 2013 Sep 17,110(38):15383-8; Proc Natl Acad Sci US A. 2015 Jan 6,112(1):202-7, all of which are incorporated by reference.

[0180] The HLA-peptide antigen may be isolated and / or in a substantially pure form. For example, the HLA-peptide antigen may be isolated from its natural environment or may be produced by a technological process. In some cases, the HLA-peptide antigen is provided in a form that is substantially free of other peptides or proteins.

[0181] The HLA-peptide antigen may be provided in a soluble form, or optionally, as a recombinant HLA-peptide antigen complex. Those skilled in the art may use any suitable method for producing and purifying recombinant HLA-peptide antigens. Suitable methods include, for example, the use of E. coli expression systems, insect cells, etc. Other methods include, for example, synthetic production using cell-free systems. An exemplary suitable cell-free system is described in WO2017089756, the entire contents of which are incorporated by reference.

[0182] Also provided herein are compositions comprising HLA-peptide antigens.

[0183] In some cases, the composition comprises an HLA-peptide target bound to a solid support. Examples of solid supports include, but are not limited to, beads, wells, membranes, tubes, columns, plates, Sepharose, magnetic beads, and chips. Exemplary solid supports are described, for example, in Catalysts 2018, 8, 92; doi:10.3390 / catal8020092, the entire contents of which are incorporated by reference.

[0184] The HLA-peptide antigen can be bound to the solid support by any suitable method known in the art. Optionally, the HLA-peptide antigen is covalently bound to the solid support.

[0185] In some cases, the HLA-peptide antigen is bound to the solid support via an affinity binding pair. Affinity binding pairs generally involve specific interactions between two molecules. A ligand having affinity for its binding partner molecule may be covalently bound to the solid support, and the ligand is then used as a bait for immobilization. Common affinity binding pairs include, for example, streptavidin and biotin, avidin and biotin, and polyhistidine tags with metal ions such as copper, nickel, zinc, and cobalt. Thus, the present specification provides a composition comprising the HLA-peptide antigen disclosed herein, wherein the HLA-peptide antigen is covalently bound to an affinity tag.

[0186] The HLA-peptide antigen may comprise a detectable label. In some embodiments, the HLA-peptide antigen is complexed with a detectable label. In some embodiments, the detectable label comprises a β2-microglobulin binding molecule, for example, a labeled antibody, for example, a fluorochrome-labeled antibody.

[0187] Also referred to herein is a pharmaceutical composition comprising an HLA-peptide antigen.

[0188] The composition containing the HLA-peptide target may be a pharmaceutical composition. Such a composition may contain multiple HLA-peptide antigens. Exemplary pharmaceutical compositions are described herein. The composition may be capable of eliciting an immune response. The composition may also include an adjuvant. Examples of suitable adjuvants include, but are not limited to, 1018 ISS, alum, aluminum salts, Amplivax, AS15, BCG, CP-870, 893, CpG7909, CyaA, dSLIM, GM-CSF, IC30, IC31, imiquimod, ImuFact IMP321, IS patch, ISS, ISCOMATRIX, JuvImmune, LipoVac, MF59, monophosphoryl lipid A, Montanide IMS 1312, Montanide ISA 206, and Montanide ISA. Adjuvants include 50V, Montanide ISA-51, OK-432, OM-174, OM-197-MP-EC, ONTAK, PepTel vector system, PLG microparticles, resiquimod, SRL172, virosomes and other virus-like particles, YF-17D, VEGF trap, R848, β-glucan, Pam3Cys, and Aquila's QS21 Stimulon (Aquillia Biotech, Worcester, Mass., USA), which contains saponins, mycobacterial extracts, synthetic bacterial cell wall mimics, and other proprietary adjuvants, such as those derived from Ribi's Detox, Quill, or Superfos. Adjuvants such as incomplete Freund's or GM-CSF are also useful. Several immunological adjuvants specific for dendritic cells and their preparation have been previously described (e.g., MF59). Cytokines may also be used.Several cytokines are directly involved in influencing dendritic cell migration to lymphoid tissues (e.g., TNF-α), promoting the maturation of dendritic cells into efficient antigen-presenting cells for T lymphocytes (e.g., GM-CSF, IL-1, and IL-4) (U.S. Pat. No. 5,849,589, specifically incorporated herein by reference in its entirety), and acting as immune adjuvants (e.g., IL-12) (Gabrilovich DI, et al., J Immunother Emphasis Tumor Immunol. 1996(6):414-418). Additionally, the surface expression of intracellular proteins and the processing of peptides into HLA for presentation on HLA can be enhanced by interferon-γ (IFN-γ). See, e.g., York IA, Goldberg AL, Mo XY, Rock KL. Proteolysis and class I major histocompatibility complex antigen presentation. Immunol Rev. 1999;172:49-66; and Rock KL, Goldberg AL. Degradation of cell proteins and the generation of MHC class I-presented peptides. Ann Rev Immunol. 1999;17:12. 739-779, all of which are incorporated by reference.

[0189] Also provided herein are host cells comprising the HLA-peptide antigens disclosed herein. In some embodiments, the host cells comprise a polynucleotide encoding an HLA-restricted peptide defined by the HLA-peptide antigen. In some embodiments, the polynucleotide is heterologous to the host cell. In some embodiments, the host cell does not contain endogenous MHC. In some embodiments, the host cell comprises exogenous HLA class I molecules. In some embodiments, the host cell is a K562 or A375 cell. In some embodiments, the host cell is a cultured cell derived from a tumor cell line. In some embodiments, the tumor cell line expresses the HLA subtype defined by the HLA-peptide antigen.

[0190] Also provided herein is a cell culture system comprising the host cells and cell culture medium disclosed herein. In some embodiments, the host cells express an HLA class I subtype defined by an HLA-peptide antigen, and the cell culture medium comprises a restricted peptide defined by the HLA-peptide antigen.

[0191] ABP Also provided herein are ABPs that specifically bind to the HLA-peptide antigens disclosed herein. In some embodiments, the ABPs disclosed herein specifically bind to HLA-peptide neoantigens comprising an HLA-restricted peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, wherein the HLA class I molecule and the HLA-restricted peptide are each selected from the HLA-peptide neoantigens set forth in any one of SEQ ID NOS: 10,755-29,364, and the ABP comprises a TCR or an antigen-binding fragment thereof. For example, in the case of the ABPs disclosed herein, the targets of the ABP are an HLA class I molecule and an associated HLA-restricted peptide, each selected from a single HLA-peptide neoantigen set forth in any one of the foregoing SEQ ID NOS: i.e., the HLA class I molecule and the HLA-restricted peptide are each selected from the same SEQ ID NOS. For example, an ABP targeting SEQ ID NO: 19865 will bind to HLA-A*11:01 in a complex with a restriction peptide of the sequence VVVGADGVGK.

[0192] HLA-peptide neoantigens can be expressed on the surface of any suitable target cell, including tumor cells.

[0193] In some embodiments, the ABP specifically binds to a complex comprising HLA and an HLA-restricted peptide (HLA-peptide), e.g., derived from a tumor. In some embodiments, the ABP does not bind to HLA in the absence of the HLA-restricted peptide. In some embodiments, the ABP binds to tumor cells that present human MHC in complex with an HLA-restricted peptide, optionally the HLA-restricted peptide being a tumor antigen that characterizes the cancer. In some aspects, the ABP binds to a complex comprising HLA and an HLA-restricted peptide when naturally presented on a cell, such as a tumor cell.

[0194] ABPs can bind to each portion of the HLA-peptide complex (i.e., the HLA and the peptide representing each portion of the complex), and when bound together, create new targets and protein surfaces for interaction and binding with the ABP, distinct from the surfaces presented by the peptide alone or the HLA subtype alone. Generally, the new targets and protein surfaces formed by HLA binding to the peptide do not exist in the absence of the portions of the HLA-peptide complex. In some embodiments, the ABP binds to the HLA-peptide neoantigen through at least one contact point with an HLA class I molecule and at least one contact point with the HLA-restricted peptide.

[0195] In some embodiments, the ABPs provided herein modulate binding of an HLA-peptide to one or more ligands of the HLA-peptide.

[0196] In more specific embodiments, the ABP specifically binds to a neo-antigen listed in Table 5A. In more specific embodiments, the ABP specifically binds to a neo-antigen listed in Table 5B. In more specific embodiments, the ABP specifically binds to a neo-antigen listed in Table 6. In more specific embodiments, the ABP specifically binds to a neo-antigen listed in Table 7.

[0197] In some embodiments of the ABP, the HLA-restricted peptide comprises a RAS mutation. In some embodiments of the ABP, the RAS mutation is a RAS G12 mutation. RAS may be KRAS, NRAS, or HRAS. In some embodiments of the ABP, the HLA-restricted peptide comprises a RAS G12 mutation. In some embodiments of the ABP, the HLA-restricted peptide comprises an NRAS G12 mutation. In some embodiments of the ABP, the HLA-restricted peptide comprises an HRAS G12 mutation. Because amino acid positions 1-50 of HRAS, KRAS, and NRAS are identical, one skilled in the art will understand that an HLA class I-restricted peptide comprising a RAS G12 mutation corresponds to KRAS G12, NRAS G12, and HRAS G12 mutations. By way of example only, SEQ ID NO: 14954, described as a KRAS G12C neoantigen, and SEQ ID NO: 14955, described as a NRAS G12C neoantigen, both have the same HLA-peptide pair (HLA-A*02:01_KLVVVGACGV). Thus, SEQ ID NOs: 14954 and 14955 describe the same KRAS / NRAS / HRAS G12C HLA-peptide neoantigen.

[0198] In some embodiments, the G12 mutation is a G12C, G12D, G12V, or G12A mutation. In some embodiments, the HLA-restricted peptide comprises a RAS G12 mutation and the HLA class I molecule is selected from HLA-A*02:01, HLA-A*11:01, HLA-A*31:01, HLA-C*01:02, and HLA-A*03:01.

[0199] In certain embodiments of the ABP, the HLA-peptide neoantigen is selected from the following: a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide Lys Leu Val Val Val Gly Ala Cys Gly Val; a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide Val Val Val Gly Ala Asp Gly Val Gly Lys; a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide Val Val Gly Ala Asp Gly Val Gly Lys; a RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide Val Val Val Gly Ala Val Gly Val Gly Lys; and a RAS_G12V MHC class I antigen comprising HLA-A*31:01 and the restriction peptide Val Val Val Gly Ala Val Gly Val Gly Lys. MHC class I antigens; RAS_G12V MHC class I antigens comprising HLA-A*11:01 and the restriction peptide Val Val Gly Ala Val Gly Val Gly Lys; RAS_G12V MHC class I antigens comprising HLA-C*01:02 and the restriction peptide Ala Val Gly Val Gly Lys Ser Ala Leu; and RAS_G12V MHC class I antigens comprising HLA-A*03:01 and the restriction peptide Val Val Val Gly Ala Val Gly Val Gly Lys.

[0200] In some embodiments of the ABP, the HLA-peptide neoantigen is selected from the following: a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide Lys Leu Val Val Val Gly Ala Cys Gly Val; a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide Val Val Val Gly Ala Asp Gly Val Gly Lys; a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide Val Val Gly Ala Asp Gly Val Gly Lys; a RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide Val Val Val Gly Ala Val Gly Val Gly Lys; or a RAS_G12V MHC class I antigen comprising HLA-A*31:01 and the restriction peptide Val Val Val Gly Ala Val Gly Val Gly Lys. MHC class I antigens; RAS_G12V MHC class I antigens comprising HLA-A*11:01 and the restriction peptide Val Val Gly Ala Val Gly Val Gly Lys; RAS_G12V MHC class I antigens comprising HLA-C*01:02 and the restriction peptide Ala Val Gly Val Gly Lys Ser Ala Leu; and RAS_G12V MHC class I antigens comprising HLA-A*03:01 and the restriction peptide Val Val Val Gly Ala Val Gly Val Gly Lys.

[0201] In some embodiments of the ABP, the HLA-peptide neoantigen is selected from: a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide LysLeuValValValGlyAlaCysGlyVal; a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide ValValValGlyAlaAspGlyValGlyLys; and a RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide ValValValGlyAlaValGlyValGlyLys. In some embodiments of the ABP, the antigen comprises HLA-A*02:01 and the restriction peptide LysLeuValValValGlyAlaCysGlyVal. In some embodiments of the ABP, the antigen comprises HLA-A*11:01 and the restriction peptide ValValValGlyAlaAspGlyValGlyLys. In some embodiments of the ABP, the antigen comprises HLA-A*11:01 and the restriction peptide Val Val Val Gly Ala Val Gly Val Gly Lys.

[0202] In some embodiments of an ABP that binds to a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the constraint peptide LysLeuValValGlyAlaCysGlyVal, the ABP binds to such a RAS_G12 MHC class I antigen with higher affinity than a RAS_G12C MHC class I antigen comprising the constraint peptide LysLeuValValGlyAlaCysGlyVal and a different HLA subtype. In some embodiments, the ABP binds to such a RAS_G12 MHC class I antigen with higher affinity than a RAS_G12 MHC class I antigen comprising the constraint peptide LysLeuValValGlyAlaCysGlyVal and a different HLA-A2 subtype. In some embodiments, the ABP does not bind to a RAS_G12C MHC class I antigen comprising the constraint peptide LysLeuValValGlyAlaCysGlyVal and a different HLA-A2 subtype.

[0203] In some embodiments of ABPs that bind to antigens containing a specific RAS G12 mutation, the ABP does not bind to the specific antigen with lower affinity than antigens containing a different RAS G12 mutation. For example, an ABP that binds to a RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide LysLeuValValGlyAlaCysGlyVal does not bind to that RAS_G12C MHC class I antigen with lower affinity than antigens containing a different RAS G12 mutation. In some embodiments of ABPs that bind to antigens containing a specific RAS G12 mutation, the ABP binds to the specific antigen with higher affinity than antigens containing a different RAS G12 mutation. For example, an ABP that binds to a RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide LysLeuValValGlyAlaCysGlyVal may bind to the RAS_G12C MHC class I antigen with higher affinity than antigens containing a different RAS G12 mutation. In some embodiments, an ABP binds to a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide Lys Leu Val Val Val Gly Ala Cys Gly Val with higher affinity than to an antigen comprising the restriction peptide KLVVVGAVGV and an HLA-A2 molecule. In certain embodiments, such an ABP does not bind to an antigen comprising the restriction peptide KLVVVGAVGV and an HLA-A2 molecule.

[0204] In some embodiments, the higher affinity is at least 2-fold, at least 5-fold, or at least 10-fold.

[0205] Affinity differences can be determined by any means known in the art, hi some embodiments, such affinity differences are assessed by MSD-ECL, SPR, BLI, or flow cytometry.

[0206] In some embodiments, the ABP is an ABP that competes with the exemplary ABPs provided herein. In some aspects, the ABP that competes with the exemplary ABPs provided herein binds to the same epitope as the exemplary ABPs provided herein.

[0207] In some embodiments, the ABPs described herein are referred to herein as "variants." In some embodiments, variants are derived from any of the sequences provided herein with one or more conservative amino acid substitutions. Conservative amino acid substitutions are described herein. In preferred embodiments, the non-conservative amino acid substitutions do not interfere with or inhibit the biological activity of the functional variant. In even more preferred embodiments, the non-conservative amino acid substitutions enhance the biological activity of the functional variant, such that the biological activity of the functional variant is improved compared to the parent ABP.

[0208] TCR In one aspect, an ABP provided herein, e.g., an ABP that specifically binds to an HLA-peptide target disclosed herein, comprises a T cell receptor (TCR). The TCR may be isolated and purified.

[0209] In the majority of T cells, the TCR is a heterodimeric polypeptide consisting of an α (α) chain and a β (β) chain, encoded by TRA and TRB, respectively. The α chain generally includes an α variable region encoded by TRAV, an α joining region encoded by TRAJ, and an α constant region encoded by TRAC. The β chain generally includes a β variable region encoded by TRBV, a β diversity region encoded by TRBD, a β joining region encoded by TRBJ, and a β constant region encoded by TRBC. The TCR-α chain is generated by V-J recombination of the αV and J segments, while the β chain receptor is generated by V(D)J recombination of the βV, D, and J segments. Additional TCR diversity results from junctional diversity. Within each junction, several bases (called N and P nucleotides) can be deleted or added. In a minority of T cells, the TCR includes a γ chain and a δ chain. The TCR γ chain is generated by V(D)J rearrangement, and the TCR δ chain is generated by V(D)J rearrangement (Kenneth Murphy, Paul Travers, and Mark Walport, Janeway's Immunology 7th edition, Garland Science, 2007, incorporated by reference in its entirety). The antigen-binding site of a TCR generally comprises six complementarity-determining regions (CDRs). The α chain contributes three CDRs: alpha ("α") CDR1, αCDR2, and αCDR3. Similarly, the β chain contributes three CDRs: beta ("β") CDR1, βCDR2, and βCDR3. In general, αCDR3 and βCDR3 are the regions most affected by V(D)J rearrangement and account for much of the variation in the TCR repertoire.

[0210] The TCR can specifically recognize an HLA-peptide target, such as those disclosed in Table 7, Table A, AACR GENIE results, or SEQ ID NOs: 29358-29364 (SEQ ID NOs: 10,755-29,364) described herein. Therefore, the TCR can be an ABP that specifically binds to an HLA-peptide. The TCR can be soluble, similar to antibodies secreted by B cells, for example. Alternatively, the TCR can be membrane-bound on cells such as T cells or natural killer (NK) cells. Therefore, the TCR can be used in contexts corresponding to soluble antibodies and / or membrane-bound CARs.

[0211] Any TCR disclosed herein may comprise an alpha variable ("V") segment, an alpha joining ("J") segment, optionally an alpha constant region, a beta variable ("V") segment, optionally a beta diversity ("D") segment, a beta joining ("J") segment, and optionally a beta constant region.

[0212] In some embodiments, the TCR or CAR is a recombinant TCR or CAR. The recombinant TCR or CAR may comprise any of the TCRs identified herein, but also include one or more modifications. Exemplary modifications, such as amino acid substitutions, are described herein. The amino acid substitutions described herein can be made with reference to the IMGT nomenclature and amino acid numbering, as referenced at www.imgt.org.

[0213] The recombinant TCR or CAR may be a human TCR or CAR that contains a fully human sequence, e.g., a naturally occurring human sequence. The recombinant TCR or CAR may retain its naturally occurring human variable domain sequence, but include modifications to the alpha constant region, the beta constant region, or both the alpha and beta constant regions. Such modifications to the TCR constant region may improve TCR assembly and expression for TCR gene therapy, for example, by driving preferential pairing of exogenous TCR chains.

[0214] In some embodiments, the α and β constant regions are modified by replacing the murine constant region sequences with the entire human constant region sequences. Such "murinized" TCRs and methods for making them are described in Cancer Res. 2006 Sep 1;66(17):8878-86, which is incorporated by reference in its entirety.

[0215] In some embodiments, the α and β constant regions are modified by replacing specific human residues with mouse residues (human-to-mouse amino acid exchange), making one or more amino acid substitutions in the human TCR α constant (TRAC) region, TCR β constant (TRBC) region, or TRAC and TRAB regions. The one or more amino acid substitutions in the TRAC region can include a Ser substitution at residue 90, an Asp substitution at residue 91, a Val substitution at residue 92, a Pro substitution at residue 93, or any combination thereof. The one or more amino acid substitutions in the human TRBC region can include a Lys substitution at residue 18, an Ala substitution at residue 22, an Ile substitution at residue 133, a His substitution at residue 139, or any combination thereof. Such targeted amino acid substitutions are described in J Immunol June 1, 2010, 184 (11) 6223-6231, the entire contents of which are incorporated by reference.

[0216] In some embodiments, human TRAC contains an Asp substitution at residue 210, and human TRBC contains a Lys substitution at residue 134. Such substitutions may facilitate salt bridge formation between the α and β chains and TCR interchain disulfide bond formation. These targeted substitutions are described in J Immunol June 1, 2010, 184 (11) 6232-6241, which is incorporated by reference in its entirety.

[0217] In some embodiments, the human TRAC and human TRBC regions are modified to contain an introduced cysteine ​​that can improve preferential pairing of exogenous TCR chains by forming an additional disulfide bond. For example, human TRAC can contain a Cys substitution at residue 48, and human TRBC can contain a Cys substitution at residue 57. This is described in Cancer Res. 2007 Apr 15;67(8):3898-903 and Blood. 2007 Mar 15;109(6):2331-8, which are incorporated by reference in their entireties.

[0218] In recombinant TCRs or CARs, the α and β chains may also contain other modifications.

[0219] In some embodiments, the α and β chains are modified by linking the extracellular domains of the α and β chains to a complete human CD3ζ (CD3-ζ) molecule. Such modifications are described in J Immunol June 1, 2008, 180(11)7736-7746; Gene Ther. 2000 Aug;7(16):1369-77; and Open Gene Therapy Journal, 2011, 4:11-22, which are incorporated by reference in their entirety.

[0220] In some embodiments, the transmembrane region of the alpha chain is modified by introducing hydrophobic amino acid substitutions, as described in J Immunol June 1, 2012, 188(11)5538-5546, which is incorporated by reference in its entirety.

[0221] The α or β chain can be modified by modifying any one of the N-glycosylation sites within the amino acid sequence, as described in J Exp Med. 2009 Feb 16;206(2):463-475, which is incorporated by reference in its entirety.

[0222] The α and β chains can each contain a dimerization domain, e.g., a heterologous dimerization domain. Such heterologous domains can be leucine zippers, 5H3 domains, or hydrophobic proline-rich counter domains, or other similar domains, as known in the art. In one example, the α and β chains can be modified by introducing 30-mer segments into the carboxyl termini of the α and β extracellular domains, so that the segments selectively associate to form stable leucine zippers. Such modifications are described in PNAS November 22, 1994. 91(24)11408-11412; https: / / doi.org / 10.1073 / pnas.91.24.11408, which are incorporated by reference in their entirety.

[0223] The TCRs identified herein may be modified to contain mutations that result in increased affinity or half-life, such as those described in WO2012 / 013913, which is incorporated by reference in its entirety.

[0224] The recombinant TCR or CAR may be a single-chain TCR (scTCR). Such scTCRs may include an α chain variable region sequence fused to the N-terminus of a TCR α chain constant region extracellular sequence, a TCR β chain variable region fused to the N-terminus of a TCR β chain constant region extracellular sequence, and a linker sequence connecting the C-terminus of the α segment to the N-terminus of the β segment, or vice versa. In some embodiments, the constant region extracellular sequences of the α and β segments of the scTCR are linked by a disulfide bond. In some embodiments, the length of the linker sequence and the position of the disulfide bond are such that the variable region sequences of the α and β segments are oriented relative to each other substantially similar to a native αβ T cell receptor. Exemplary scTCRs are described in U.S. Patent No. 7,569,664, incorporated by reference in its entirety.

[0225] In some cases, the variable regions of the scTCR may be covalently linked by a short peptide linker, such as those described in Gene Therapy volume 7, pages 1369-1377 (2000). The short peptide linker may be a serine-rich linker or a glycine-rich linker. For example, the linker may be (Gly4Ser)3, as described in Cancer Gene Therapy (2004) 11, 487-496, the entire contents of which are incorporated by reference.

[0226] The recombinant TCR or its antigen-binding fragment may be expressed as a fusion protein. For example, the TCR or its antigen-binding fragment may be fused with a toxin. Such a fusion protein is described in Cancer Res. 2002 Mar 15;62(6):1757-60. The TCR or its antigen-binding fragment may be fused with the Fc region of an antibody. Such a fusion protein is described in J Immunol May 1, 2017, 198(1 Supplement)120.9.

[0227] The antigen recognition domain of a receptor, such as a TCR or CAR, can be linked to one or more intracellular signaling components, such as signaling components. These intracellular signaling components mimic activation through an antigen receptor complex, such as a TCR complex, and / or a signal through another cell surface receptor. For example, an HLA-peptide-specific binding component (e.g., an ABP, such as a TCR) can be linked to one or more transmembrane and intracellular signaling domains. In some embodiments, the transmembrane domain is fused to the extracellular domain. In one embodiment, a transmembrane domain that naturally associates with one of the domains in the receptor, e.g., a CAR, is used. In some instances, the transmembrane domain is selected or modified by amino acid substitution to avoid such domains being bound to the transmembrane domain of the same or a different surface membrane protein and minimize interaction with other members of the receptor complex.

[0228] In some embodiments, the transmembrane domain is derived from either a natural or synthetic source. If the source is natural, the domain in some aspects is derived from a membrane-bound or transmembrane protein. Transmembrane regions include those derived from the α, β, or ζ chain (i.e., comprising at least the transmembrane region(s)) of the T cell receptor, CD28, CD3γ, CD45, CD4, CD5, CDS, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, or CD154. Alternatively, the transmembrane domain in some embodiments is synthetic. In some aspects, synthetic transmembrane domains are dominated by hydrophobic residues such as leucine and valine. In some aspects, triplets of phenylalanine, tryptophan, and valine are found at both ends of the synthetic transmembrane domain. In some embodiments, the linkage is via a linker, spacer, and / or transmembrane domain(s).

[0229] Prominent intracellular signaling domains are those that mimic or approximate signaling through natural antigen receptors, through such receptors in combination with costimulatory receptors, and / or through costimulatory receptors alone. In some embodiments, a short oligo- or polypeptide linker, e.g., a linker 2-10 amino acids in length, such as one containing glycine and serine, e.g., a glycine-serine doublet, is present between the transmembrane domain and the cytoplasmic signaling domain of the receptor to form the link.

[0230] A receptor, e.g., a TCR or CAR, can comprise at least one intracellular signaling component. In some embodiments, the receptor comprises an intracellular component of the TCR complex, such as the TCR CD3 chain, e.g., the CD3ζ chain, which mediates T cell activation and cytotoxicity. For example, an HLA-peptide-binding ABP (e.g., a TCR or CAR) is linked to one or more cell signaling modules. In some embodiments, the cell signaling modules include a CD3 transmembrane domain, a CD3 intracellular signaling domain, and / or other CD transmembrane domains. In some embodiments, the receptor, e.g., a TCR or CAR, further comprises one or more additional molecules, e.g., a portion of Fc receptor-γ, CD8, CD4, CD25, or CD16. For example, in some aspects, the TCR or CAR comprises a chimeric molecule between CD3ζ or Fc receptor γ and CD8, CD4, CD25, or CD16.

[0231] In some embodiments, upon ligation of the TCR or CAR, the cytoplasmic domain or intracellular signaling domain of the receptor activates at least one of the normal effector functions or responses of an immune cell, e.g., a T cell engineered to express the receptor. For example, in some situations, the receptor induces a T cell function, e.g., cytolytic activity or T helper activity, such as secretion of cytokines or other factors. In some embodiments, a truncated portion of the intracellular signaling domain of an antigen receptor component or costimulatory molecule is used in place of an intact immunostimulatory chain, e.g., if it transmits an effector function signal. In some embodiments, the intracellular signaling domain(s) include the cytoplasmic sequence of a T cell receptor (TCR), and in some aspects, the cytoplasmic sequence of a co-receptor that, in its natural environment, acts in concert with such receptor to initiate signal transduction following antigen receptor binding, and / or any derivative or variant of such molecule, and / or any synthetic sequence having the same functional capability.

[0232] In the context of natural TCRs, full activation generally requires not only TCR-mediated signaling but also a costimulatory signal. Thus, in some embodiments, the receptor also contains components for generating a secondary or costimulatory signal to promote full activation. In other embodiments, the receptor does not contain components for generating a costimulatory signal. In some aspects, additional receptors are expressed in the same cell and provide components for generating a secondary or costimulatory signal.

[0233] In some embodiments, T cell activation has been described as being mediated by two classes of cytoplasmic signaling sequences: those that initiate antigen-dependent primary activation via the TCR (primary cytoplasmic signaling sequences), and those that act antigen-independently to provide secondary or costimulatory signals (secondary cytoplasmic signaling sequences). In some embodiments, a receptor contains one or both of these signaling components.

[0234] In some aspects, the receptor comprises a primary cytoplasmic signaling sequence that regulates primary activation of the TCR complex. Primary cytoplasmic signaling sequences that act in a stimulatory manner may comprise signaling motifs known as immunoreceptor tyrosine-based activation motifs, or ITAMs. Examples of ITAMs that comprise primary cytoplasmic signaling sequences include those derived from TCR or CD3ζ, FcRγ, FcRβ, CD3γ, CD3δ, CD3ε, CDS, CD22, CD79a, CD79b, and CD66d. In some embodiments, the cytoplasmic signaling molecule(s) of the CAR comprise a cytoplasmic signaling domain, a portion thereof, or a sequence derived from CD3ζ.

[0235] In some embodiments, the receptor comprises the signaling domain and / or transmembrane portion of a costimulatory receptor such as CD28, 4-1BB, OX40, DAP10, and ICOS. In some aspects, the same receptor comprises both an activating component and a costimulatory component.

[0236] In some embodiments, the activation domain is contained in one receptor, but the costimulatory component is provided by another receptor that recognizes a different antigen. In some embodiments, the receptor includes an activating or stimulatory receptor and a costimulatory receptor, both of which are expressed in the same cell (see WO2014 / 055668). The HLA-peptide targeting receptor is a stimulatory or activation receptor in some aspects, and a costimulatory receptor in other aspects. In some embodiments, the cell further includes an inhibitory receptor (e.g., iCAR, see Fedorov et al., Sci. Transl. Medicine, 5(215) (December, 2013)), e.g., a receptor that recognizes an antigen other than an HLA-peptide, thereby attenuating or inhibiting the activation signal delivered via the HLA-peptide targeting receptor by binding of the inhibitory receptor to its ligand, e.g., reducing off-target effects.

[0237] In certain embodiments, the intracellular signaling domain comprises a CD28 transmembrane and signaling domain linked to a CD3 (e.g., CD3-zeta) intracellular domain, hi some embodiments, the intracellular signaling domain comprises a chimeric CD28 and CD137 (4-1BB, TNFRSF9) costimulatory domain linked to a CD3zeta intracellular domain.

[0238] In some embodiments, the receptor includes one or more, e.g., two or more, costimulatory domains and an activation domain, e.g., a primary activation domain, in the cytoplasmic portion. Exemplary receptors include the intracellular components of CD3ζ, CD28, and 4-1BB.

[0239] In some embodiments, the CAR (or other antigen receptor, such as a TCR) further comprises a marker, such as a cell surface marker. This marker can be used to transduce or engineer cells to confirm expression of the receptor, including, for example, truncated forms of the cell surface receptor, such as truncated EGFR (tEGFR). In some aspects, the marker includes all or a portion (e.g., truncated forms) of CD34, nerve growth factor receptor (NGFR), or epidermal growth factor receptor (e.g., tEGFR). In some embodiments, the nucleic acid encoding the marker is operably linked to a polynucleotide encoding a cleavable linker sequence or a ribosomal skip sequence, such as T2A. See WO2014031687. In some embodiments, two proteins may be expressed from the same construct by introducing constructs encoding the CAR and EGFRt separated by a T2A ribosomal switch. EGFRt can be used as a marker to detect cells expressing such a construct. In some embodiments, the marker, and optional linker sequence, can be any of those disclosed in Patent Application Publication No. WO 2014031687. For example, the marker can be a truncated EGFR (tEGFR) optionally linked to a linker sequence, such as a T2A ribosomal skip sequence.

[0240] In some embodiments, the marker is a molecule, e.g., a cell surface protein, that is not naturally found within or on the surface of a T cell or part of a T cell.

[0241] In some embodiments, the molecule is a non-self molecule, eg, a non-self protein, that is not recognized as "self" by the immune system of the host into which the cells are adoptively transferred.

[0242] In some embodiments, the marker serves as a marker for genetic engineering, e.g., selection of successfully engineered cells, but does not otherwise serve any therapeutic function and / or exert any efficacy. In other embodiments, the marker may be a therapeutic molecule or a molecule that exerts some other desired effect, e.g., a ligand for cells encountered in vivo, e.g., a costimulatory or immune checkpoint molecule to enhance and / or suppress the response of cells upon adoptive transfer and encounter with the ligand.

[0243] A TCR or CAR may include one or more modified synthetic amino acids in place of one or more naturally occurring amino acids. Exemplary modified amino acids include, but are not limited to, aminocyclohexanecarboxylic acid, norleucine, alpha-amino n-decanoic acid, homoserine, S-acetylaminomethylcysteine, trans-3- and trans-4-hydroxyproline, 4-aminophenylalanine, 4-nitrophenylalanine, 4-chlorophenylalanine, 4-carboxyphenylalanine, (3-phenylserine, 3-hydroxyphenylalanine, phenylglycine, alpha-naphthylalanine, cyclohexylalanine, cyclohexylglycine, indoline-2- Carboxylic acid, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, aminomalonic acid, aminomalonic acid monoamide, N'-benzyl-N'-methyl-lysine, N',N'-dibenzyl-lysine, 6-hydroxylysine, ornithine, α-aminocyclopentanecarboxylic acid, α-aminocyclohexanecarboxylic acid, α-aminocycloheptanecarboxylic acid, α(2-amino-2-norbornane)-carboxylic acid, α,γ-diaminobutyric acid, α,γ-diaminopropionic acid, homophenylalanine, and α-tertbutylglycine.

[0244] In some embodiments, CARs are referred to as first-, second-, and / or third-generation CARs. In some embodiments, first-generation CARs are CARs that provide only a CD3 chain-inducing signal upon antigen binding. In some embodiments, second-generation CARs are those that provide such a signal as well as a costimulatory signal, for example, those that include an intracellular signaling domain from a costimulatory receptor such as CD28 or CD137. In some embodiments, third-generation CARs are CARs that include multiple costimulatory domains from different costimulatory receptors.

[0245] In some embodiments, the chimeric antigen receptor comprises an extracellular portion comprising a TCR or fragment described herein. In some aspects, the chimeric antigen receptor comprises an extracellular portion comprising a TCR or fragment described herein and an intracellular signaling domain. In some embodiments, the intracellular domain comprises an ITAM. In some aspects, the intracellular signaling domain comprises the signaling domain of the zeta chain of the CD3-zeta (CD3) chain. In some embodiments, the chimeric antigen receptor comprises a transmembrane domain connecting the extracellular domain and the intracellular signaling domain.

[0246] In some aspects, the transmembrane domain comprises the transmembrane portion of CD28. The extracellular domain and the transmembrane may be directly or indirectly linked. In some embodiments, the extracellular domain and the transmembrane are linked by a spacer as described herein. In some embodiments, the chimeric antigen receptor comprises the intracellular domain of a T cell costimulatory molecule, such as between the transmembrane domain and the intracellular signaling domain. In some aspects, the T cell costimulatory molecule is CD28 or 41BB.

[0247] In some embodiments, the CAR comprises a TCR, e.g., a TCR fragment, a transmembrane domain that is or comprises the transmembrane portion of CD28 or a functional variant thereof, and an intracellular signaling domain that comprises the signaling portion of CD28 or a functional variant thereof and the signaling portion of CD3ζ or a functional variant thereof. In some embodiments, the CAR comprises a TCR, e.g., a TCR fragment, a transmembrane domain that is or comprises the transmembrane portion of CD28 or a functional variant thereof, and an intracellular signaling domain that comprises the signaling portion of 4-1BB or a functional variant thereof and the signaling portion of CD3ζ or a functional variant thereof. In some such embodiments, the receptor further comprises a spacer that comprises a portion of an Ig molecule, such as a hinge-only spacer, e.g., a human Ig molecule, such as an Ig hinge, e.g., an IgG4 hinge.

[0248] In some embodiments, the transmembrane domain of a receptor, e.g., a CAR, is the transmembrane domain of human CD28 or a variant thereof, e.g., the 27 amino acid transmembrane domain of human CD28 (Accession Number: P10747.1).

[0249] In some embodiments, the chimeric antigen receptor comprises the intracellular domain of a T cell costimulatory molecule. In some aspects, the T cell costimulatory molecule is CD28 or 41BB.

[0250] In some embodiments, the intracellular signaling domain comprises the intracellular costimulatory signaling domain of human CD28 or a functional variant or portion thereof, e.g., the 41 amino acid domain thereof and / or such a domain having an LL to GG substitution at positions 186-187 of the native CD28 protein. In some embodiments, the intracellular domain comprises the intracellular costimulatory signaling domain of 41BB or a functional variant or portion thereof, e.g., the 42 amino acid cytoplasmic domain of human 4-1BB (Accession No.: Q07011.1) or a functional variant or portion thereof.

[0251] In some embodiments, the intracellular signaling domain comprises a human CD3ζ stimulatory signaling domain or a functional variant thereof, such as the 112AA cytoplasmic domain of human CD3ζ isoform 3 (Accession Number: P20963.2) or a CD3ζ signaling domain, as described in U.S. Pat. No. 7,446,190 or U.S. Pat. No. 8,911,993.

[0252] In some aspects, the spacer comprises only the hinge region of an IgG, e.g., only an IgG4 or IgG1 hinge. In other embodiments, the spacer is an Ig hinge linked to the CH2 and / or CH3 domains, e.g., an IgG4 hinge. In some embodiments, the spacer is an Ig hinge linked to the H2 and CH3 domains, e.g., an IgG4 hinge. In some embodiments, the spacer is an Ig hinge linked to the CH3 domain only, e.g., an IgG4 hinge. In some embodiments, the spacer is or comprises a glycine-serine rich sequence or other flexible linker, e.g., a known flexible linker.

[0253] For example, in some embodiments, a CAR comprises a TCR or a fragment thereof, e.g., any HLA-peptide specific TCR, a spacer such as an optional Ig hinge-containing spacer, a CD28 transmembrane domain, a CD28 intracellular signaling domain, and a CD3ζ signaling domain. In some embodiments, a CAR comprises a TCR or a fragment thereof, e.g., any HLA-peptide specific TCR, a spacer such as an optional Ig hinge-containing spacer, a CD28 transmembrane domain, a CD28 intracellular signaling domain, and a CD3ζ signaling domain.

[0254] Nucleotides, Vectors, Host Cells, and Related Methods Also provided are isolated nucleic acids encoding the ABPs or antigens disclosed herein, vectors containing the nucleic acids, and host cells containing the vectors and nucleic acids, as well as recombinant techniques for producing the ABPs.

[0255] Nucleic acids may be recombinant. Recombinant nucleic acids can be constructed outside of living cells by joining natural or synthetic nucleic acid segments to a nucleic acid molecule capable of replicating within a living cell, or the product of that replication. For purposes herein, replication can be in vitro or in vivo.

[0256] For recombinant production of an ABP, the nucleic acid(s) encoding the ABP may be isolated and inserted into a replicable vector for further cloning (i.e., amplification of the DNA) or expression. In some embodiments, the nucleic acid may be generated by homologous recombination, as described, for example, in U.S. Patent No. 5,204,244, which is incorporated by reference in its entirety.

[0257] Many different vectors are known in the art. The vector components generally include one or more of the following: a signal sequence, an origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence, as described, for example, in U.S. Patent No. 5,534,615, which is incorporated by reference in its entirety.

[0258] Exemplary vectors or constructs suitable for expressing an ABP, e.g., a CAR, or an antigen-binding fragment thereof, include, for example, the pUC series (Fermentas Life Sciences), the pBluescript series (Stratagene, LaJolla, CA), the pET series (Novagen, Madison, WI), the pGEX series (Pharmacia, The PFies), and the pEX series (Clontech, Palo Alto, CA). Bacteriophage vectors such as AGT10, AGT11, AZapII (Stratagene), AEMBL4, and ANM1149 are also suitable for expressing the ABPs disclosed herein.

[0259] Illustrative examples of suitable host cells are provided below. These host cells are not meant to be limiting, and any suitable host cell can be used to produce the ABPs provided herein.

[0260] Suitable host cells include prokaryotic (eg, bacterial), lower eukaryotic (eg, yeast), or higher eukaryotic (eg, mammalian) cells. Suitable prokaryotes include eubacteria, such as gram-negative or gram-positive organisms, for example, Escherichia (E. coli), Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella (S. typhimurium), Serratia (S. marcescans), Shigella, Bacillus (B. subtilis and B. licheniformis), Pseudomonas (P. aeruginosa), and Streptomyces. One useful E. coli cloning host is E. coli 294, although other strains such as E. coli B, E. coli X1776, and E. coli W3110 are also suitable.

[0261] In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for ABP-encoding vectors. Saccharomyces cerevisiae, or common baker's yeast, is a commonly used lower eukaryotic host microorganism. However, Schizosaccharomyces pombe, Kluyveromyces species (K. lactis, K. fragilis, K. bulgaricus, K. wickeramii, K. waltii, K. drosophilarum, K. thermotolerans, and K. marxianus), Yarrowia, Pichia pastoris, Candida species (C. albicans), Trichoderma reesei, Neurospora crassa, Many other genera, species, and strains of fungi are available and useful, such as S. crassa, Schwanniomyces (S. occidentalis), and filamentous fungi, e.g., Penicillium, Tolypocladium, and Aspergillus (A. nidulans and A. niger).

[0262] Useful mammalian host cells include COS-7 cells, HEK293 cells, baby hamster kidney (BHK) cells; Chinese hamster ovary (CHO); mouse Sertoli cells; African green monkey kidney cells (VERO-76); and the like.

[0263] The host cells used to produce HLA-peptide ABPs can be cultured in a variety of media. For example, commercially available media such as Ham's F10, minimal essential medium (MEM), RPMI-1640, and Dulbecco's modified Eagle's medium (DMEM) are suitable for culturing host cells. In addition, any of the media described in Ham et al., Meth. Enz., 1979, 58:44; Barnes et al., Anal. Biochem., 1980, 102:255; and U.S. Patent Nos. 4,767,704, 4,657,866, 4,927,762, 4,560,655, and 5,122,469; or WO 90 / 03430 and WO 87 / 00195 may be used. Each of the aforementioned references is incorporated by reference in its entirety.

[0264] Any of these media can be supplemented as needed with hormones and / or other growth factors (such as insulin, transferrin, or epidermal growth factor), salts (such as sodium chloride, calcium, magnesium, and phosphate), buffers (such as HEPES), nucleotides (such as adenosine and thymidine), antibiotics, trace elements (usually defined as inorganic compounds present at final concentrations in the micromolar range), and glucose or an equivalent energy source. Any other necessary supplements can also be included at appropriate concentrations that would be known to those skilled in the art.

[0265] Culture conditions such as temperature, pH, etc. will be those previously used with the host cell selected for expression and will be apparent to one skilled in the art.

[0266] When using recombinant techniques, ABPs can be produced intracellularly in the periplasmic space or directly secreted into the medium. If ABPs are produced intracellularly, the first step is to remove particulate debris, either host cells or lysed fragments, for example, by centrifugation or ultrafiltration. Procedures for isolating ABPs secreted into the periplasmic space of E. coli are described, for example, in Carter et al. (Bio / Technology, 1992, 10:163-167), the entire contents of which are incorporated by reference. Briefly, cell paste is thawed in the presence of sodium acetate (pH 3.5), EDTA, and phenylmethylsulfonyl fluoride (PMSF) for about 30 minutes. Cell debris can be removed by centrifugation.

[0267] In some embodiments, the ABP is produced in a cell-free system. In some aspects, the cell-free system is an in vitro transcription and translation system, such as that described in Yin et al., mAbs, 2012, 4:217-225, which is incorporated by reference in its entirety. In some aspects, the cell-free system utilizes cell-free extracts derived from eukaryotic or prokaryotic cells. In some aspects, the prokaryotic cell is E. coli. Cell-free expression of the ABP can be useful, for example, when ATP accumulates in the cell as insoluble aggregates or when yields from periplasmic expression are low.

[0268] When ABPs are secreted into the medium, supernatants from such expression systems are typically first concentrated using commercially available protein concentration filters, such as Amicon® or Millipore® Pellcon® ultrafiltration units. A protease inhibitor, such as PMSF, may be included in any of the foregoing steps to inhibit proteolysis, and antibiotics may be included to prevent the growth of adventitious contaminants.

[0269] ABP compositions prepared from cells can be purified using, for example, hydroxylapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography. Affinity chromatography is a particularly useful purification technique. The suitability of protein A as an affinity ligand varies depending on the type and isotype of immunoglobulin Fc domain present in the ABP. Protein A can be used to purify ABPs containing human γ1, γ2, or γ4 heavy chains (Lindmark et al., J. Immunol. Meth., 1983, 62:1-13, incorporated by reference in its entirety). Protein G is useful for all mouse isotypes and human γ3 (Guss et al., EMBO J., 1986, 5:1567-1575, incorporated by reference in its entirety).

[0270] The matrix to which the affinity ligand is attached is most often agarose, although other matrices are available. Mechanically stable matrices such as controlled pore glass or poly(styrenedivinyl)benzene allow for faster flow rates and shorter processing times than can be achieved with agarose. H3 If the domain is involved, BakerBond ABX® resin is useful for purification.

[0271] Other techniques for protein purification, such as fractionation on ion exchange columns, ethanol precipitation, reverse-phase HPLC, chromatography on silica, chromatography on heparin Sepharose®, chromatofocusing, SDS-PAGE, and ammonium sulfate precipitation, are also available and can be applied by one skilled in the art.

[0272] After the preliminary purification step(s), the mixture containing the ABP of interest and contaminants may be subjected to low pH hydrophobic interaction chromatography, typically performed at low salt concentrations (e.g., about 0 to about 0.25 M salt), using an elution buffer at a pH of about 2.5 to about 4.5.

[0273] Method for producing HLA-peptide ABP Preparation of HLA-peptide antigens The HLA-peptide antigens used to isolate or generate the ABPs provided herein can be intact HLA-peptides or fragments of HLA-peptides, for example, in the form of isolated proteins or proteins expressed on the surface of cells.

[0274] In some embodiments, the HLA peptiantigen is a non-naturally occurring variant of an HLA-peptide, such as an HLA-peptide protein having an amino acid sequence or post-translational modification that does not occur in nature.

[0275] In some embodiments, the HLA-peptide antigen is truncated, for example, by removal of an intracellular or transmembrane sequence, or a signal sequence, hi some embodiments, the HLA-peptide antigen is fused at its C-terminus to a human IgG1 Fc domain or a polyhistidine tag.

[0276] Methods and systems for identifying ABPs ABPs that bind to HLA-peptides can be identified using any method known in the art, such as phage display, immunization of a subject, or isolation of ABP-expressing cells and subsequent sequencing of the ABP.

[0277] One method for identifying an antigen-binding protein includes providing at least one HLA-peptide target and allowing the at least one target to bind to an antigen-binding protein, thereby identifying the antigen-binding protein. The antigen-binding protein may be present in a library comprising a plurality of individual antigen-binding proteins.

[0278] In some embodiments, the library is a phage display library. The phage display library can be developed to be substantially free of antigen binding proteins that nonspecifically bind to the HLA of the HLA-peptide target. The antigen binding proteins can be present in a yeast display library that comprises a plurality of individual antigen binding proteins. The yeast display library can be developed to be substantially free of antigen binding proteins that nonspecifically bind to the HLA of the HLA-peptide target.

[0279] In some embodiments, the library is a yeast display library.

[0280] In some embodiments, the binding step is performed more than once, optionally at least three times, e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times.

[0281] Additionally, the method may also include contacting the antigen binding protein with one or more peptide-HLA complexes that are different from the HLA-peptide target to determine whether the antigen binding protein selectively binds to the HLA-peptide target.

[0282] Thus, provided herein is a system for identifying ABPs that selectively bind to one or more antigens described herein. In some embodiments, the system includes: (a) an isolated antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, the HLA-restricted peptide being located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, the antigen being selected from the antigens set forth in any one of SEQ ID NOS: 10,755-29,364; and (b) a library comprising a plurality of different antigen-binding proteins. In some embodiments, the library is a phage display library.

[0283] In some embodiments of this system, the antigen is bound to a solid support. The solid support may include, for example, beads, wells, membranes, tubes, columns, plates, sepharose, magnetic beads, cells, or chips. In some embodiments, the antigen comprises a first member of an affinity binding pair and the solid support comprises a second member of the affinity binding pair. In some embodiments, the first member is streptavidin and the second member is biotin. In some embodiments, the antigen bound to the solid support is an HLA multimer (e.g., a tetramer) comprising at least one HLA-peptide target.

[0284] In some embodiments of this system, the library (e.g., phage display library) is a human library. In some embodiments of this system, the library (e.g., phage display library) is a humanized library.

[0285] In some embodiments, the system further includes a negative control antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, where the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, and the negative control antigen comprises a different restricted peptide, a different HLA class I molecule, or a different restricted peptide and a different HLA class I molecule. In some embodiments, the negative control antigen comprises a different restricted peptide but the same HLA class I molecule as the antigen.

[0286] In some embodiments, the system includes a reaction mixture comprising an antigen and a plurality of phages from a phage display library.

[0287] Another method for identifying an antigen-binding protein may include obtaining at least one HLA-peptide target, administering the HLA-peptide target, optionally in combination with an adjuvant, to a subject (e.g., a mouse, rabbit, or llama), and isolating the antigen-binding protein from the subject. Isolating the antigen-binding protein may include screening the subject's serum to identify the antigen-binding protein. The method may also include contacting the antigen-binding protein with one or more peptide-HLA complexes different from the HLA-peptide target, for example, to determine whether the antigen-binding protein selectively binds to the HLA-peptide target. The identified antigen-binding protein may be humanized.

[0288] In some embodiments, isolating the antigen-binding protein comprises isolating T cells from a subject that express the antigen-binding protein. The T cells can be used to generate hybridomas. The T cells can also be used to clone one or more of the CDRs. The T cells can also be immortalized, for example, by using EBV transformation. The sequence encoding the antigen-binding protein can be cloned from the immortalized T cells or directly from T cells isolated from an immunized subject. A library comprising the antigen-binding proteins of T cells can also be generated, and optionally the library is phage display or yeast display.

[0289] Another method for identifying an antigen binding protein may include obtaining a cell containing the antigen binding protein, contacting the cell with an HLA multimer (e.g., a tetramer) comprising at least one HLA-peptide target, and identifying the antigen binding protein via binding between the HLA multimer and the antigen binding protein.

[0290] Another method for identifying an antigen-binding protein can include obtaining cells containing an antigen-binding protein (ABP) and determining the sequence of the ABP. For example, this method can include contacting cells with an HLA multimer (e.g., a tetramer) containing at least one HLA-peptide target; optionally isolating the cells using flow cytometry (e.g., fluorescence-activated cell sorting "FACS"), magnetic separation, or single-cell separation; and sequencing polynucleotides from the isolated cells to determine the sequence of the ABP.

[0291] In some embodiments, isolation is performed by enriching for a particular cell population by positive selection, or by depleting a particular cell population by negative selection. In some embodiments, positive or negative selection results in a relatively high level (marker) in the positively or negatively selected cells, respectively. 高 This is achieved by incubating cells with one or more antibodies or other binding agents that specifically bind to one or more surface markers expressed or expressed (marker+). For example, a population of cells known or suspected to contain T cells can be positively sorted based on binding to tetramers containing the HLA-peptide of interest (e.g., neoantigen). FACS isolation can also include removal of cells that bind to non-desired HLA-peptide targets. For example, cells can be positively sorted based on binding to tetramers containing the HLA-peptide of interest (e.g., neoantigen) or negatively sorted based on binding to tetramers containing the non-desired HLA-peptide (e.g., wild-type peptide sequence corresponding to the neoantigen of interest).

[0292] Isolation of cells expressing an ABP-containing protein (e.g., FACS-based isolation of T cells) can include isolation of cells from a subject. Cells from a subject can be isolated from a variety of biological samples, including, but not limited to, bodily fluids such as blood, plasma, serum, cerebrospinal fluid, synovial fluid, urine, and sweat, tissue, and organ samples. Biological samples can be samples obtained directly from biological sources or processed samples. The sample from which cells from a subject are derived or isolated can be blood or a blood-derived sample, or can be derived from an apheresis or leukapheresis product. Exemplary samples include whole blood, peripheral blood mononuclear cells (PBMCs), white blood cells, bone marrow, thymus, tissue biopsy, tumor, leukemia, lymphoma, lymph node, gut-associated lymphoid tissue, mucosa-associated lymphoid tissue, spleen, other lymphoid tissue, liver, lung, stomach, intestine, colon, kidney, pancreas, breast, bone, prostate, cervix, testis, ovary, tonsil, or other organ, and / or cells derived therefrom. Exemplary cells and cell populations that express ABP-containing proteins include, but are not limited to, activated T cells, tumor infiltrating lymphocytes (TILs), PBMCs, cultured (e.g., expanded) T cells, naive T (TN) cells, effector T cells (TEFF), memory T cells, stem cell memory T cells (TSCM), central memory T cells (TCM), effector memory T cells (TEM), terminally differentiated effector memory T cells, immature T cells, mature T cells, helper T cells, cytotoxic T cells, mucosal-associated invariant T (MALT) cells, regulatory T cells (Tregs), TH1 cells, TH2 cells, TH3 cells, TH17 cells, TH9 cells, TH22 cells, follicular helper T cells, natural killer T cells (NKT), alpha beta T cells, and gamma delta T cells.

[0293] Sequencing of cells expressing ABP-containing proteins can be performed by techniques known to those skilled in the art, such as the Chromium Single Cell Immune Profiling System (10x Genomics).

[0294] Another method of identifying an antigen-binding protein may include obtaining one or more cells containing the antigen-binding protein; activating the one or more cells using at least one HLA-peptide target presented on at least one antigen-presenting cell (APC); and identifying the antigen-binding protein by selecting the one or more cells activated by interaction with the at least one HLA-peptide target.

[0295] The cell can be, for example, T cell, optionally CTL or NK cell.The method can further comprise isolating the cell optionally using flow cytometry, magnetic separation or single cell separation.The method can further comprise sequencing the antigen-binding protein.

[0296] Methods for manipulating cells with ABPs Also provided are methods, nucleic acids, compositions, and kits for expressing ABPs, including receptors, including TCRs, CARs, etc., and for producing genetically engineered cells that express such ABPs. Genetic engineering generally involves introducing nucleic acids encoding the recombinant or engineered components into cells, such as by retroviral transduction, transfection, or transformation.

[0297] In some embodiments, gene transfer is achieved by first stimulating the cells with a combination of stimuli that induce a response such as proliferation, survival, and / or activation, e.g., as measured by expression of cytokines or activation markers, followed by transduction of the activated cells and expansion in culture to numbers sufficient for clinical use.

[0298] In some situations, overexpression of a stimulatory factor (e.g., a lymphokine or cytokine) can be toxic to a subject. Thus, in some situations, engineered cells include a segment that renders the cells susceptible to negative selection in vivo, such as upon administration in adoptive immunotherapy. For example, in some embodiments, the cells are engineered so that they can be eliminated as a result of changes in the in vivo conditions of the patient to whom they are administered. A negatively selectable phenotype can be generated from the insertion of a gene that confers sensitivity to an administered agent, e.g., a compound. Negative selectable genes include the herpes simplex virus type I thymidine kinase (HSV-I TK) gene (Wigler et al., Cell II:223, 1977), which confers ganciclovir sensitivity, the cellular hypoxanthine phosphoribosyltransferase (HPRT) gene, the cellular adenine phosphoribosyltransferase (APRT) gene, and bacterial cytosine deaminase (Mullen et al., Proc. Natl. Acad. Sci. USA. 89:33 (1992)).

[0299] In some embodiments, the cells are further engineered to promote the expression of cytokines or other factors. Various methods for introducing genetically engineered components, such as antigen receptors (e.g., TCRs), are well known and may be used with the provided methods and compositions. Exemplary methods include methods for introducing nucleic acids encoding receptors, including viruses, such as retroviruses or lentiviruses, transduction, transposons, nuclease-mediated gene editing (e.g., CRISPR, TALEN, meganuclease, or ZFN editing systems), and electroporation. For example, nuclease-mediated gene editing, particularly for editing T cells, is described in detail in International Applications WO / 2018 / 232356 and PCT / US2018 / 058230, which are incorporated herein by reference for all purposes.

[0300] In some embodiments, recombinant nucleic acids are introduced into cells using recombinant infectious viral particles, such as vectors derived from Simian Virus 40 (SV40), adenovirus, or adeno-associated virus (AAV). In some embodiments, recombinant nucleic acids are introduced into T cells using retroviral vectors, such as recombinant lentiviral or gamma retroviral vectors (see, e.g., Koste et al. (2014) Gene Therapy 2014 Apr. 3. doi:10.1038 / gt.2014.25; Carlens et al. (2000) Exp Hematol 28(10):1137-46; Alonso-Camino et al. (2013) Mol Ther Nucl Acids 2,e93; Park et al., Trends Biotechnol. 2011 Nov. 29(11):550-557).

[0301] In some embodiments, the retroviral vector has a long terminal repeat (LTR), such as a retroviral vector derived from Moloney murine leukemia virus (MoMLV), myeloproliferative sarcoma virus (MPSV), murine embryonic stem cell virus (MESV), murine stem cell virus (MSCV), spleen follicular fungus virus (SFFV), or adeno-associated virus (AAV). Most retroviral vectors are derived from murine retroviruses. In some embodiments, retroviruses include those derived from avian or mammalian cell sources. Retroviruses are typically amphotropic, meaning that they can infect host cells of several species, including humans. In one embodiment, the retroviral gag, pol, and / or env sequences are replaced with the gene to be expressed. Several exemplary retroviral systems have been described (e.g., U.S. Patent Nos. 5,219,740; 6,207,453; 5,219,740; Miller and Rosman (1989) BioTechniques 7:980-990; Miller, AD (1990) Human Gene Therapy 1:5-14; Scarpa et al. (1991) Virology 180:849-852; Burns et al. (1993) Proc. Natl. Acad. Sci. USA 90:8033-8037; and Boris-Lawrie and Temin (1993) Cur. Opin. Genet. Develop. 3:102-109).

[0302] Methods for lentiviral transduction are known. Exemplary methods are described, for example, in Wang et al. (2012) J. Immunother. 35(9)689-701; Cooper et al. (2003) Blood. 101:1637-1644; Verhoeyen et al. (2009) Methods Mol Biol. 506:97-114; and Cavalieri et al. (2003) Blood. 102(2)497-505).

[0303] In some embodiments, recombinant nucleic acids are transferred into T cells via electroporation (see, e.g., Chicaybam et al. (2013) PLoS ONE 8(3):e60298; Van Tedeloo et al. (2000) Gene Therapy 7(16):1431-1437, and Roth et al. (2018) Nature 559:405-409). In some embodiments, recombinant nucleic acids are introduced into T cells via transposition (see, e.g., Manuri et al. (2010) Hum Gene Ther 21(4):427-437; Sharma et al. (2013) Molec Ther Nucl Acids 2,e74, and Huang et al. (2009) Methods Mol Biol 506:115-126). Other methods for introducing and expressing genetic material into immune cells include calcium phosphate transfection (e.g., as described in Current Protocols in Molecular Biology, John Wiley & Sons, New York, NY), protoplast fusion, cationic liposome-mediated transfection; tungsten particle-facilitated particle bombardment (Johnston, Nature, 346:776-777 (1990)), and strontium phosphate DNA coprecipitation (Brash et al., Mol. Cell Biol., 7:2031-2034 (1987)).

[0304] Other approaches and vectors for introducing nucleic acids encoding recombinant products include those described in, for example, International Patent Application Publication No. WO2014055668 and U.S. Patent No. 7,446,190.

[0305] Among the additional nucleic acids, e.g., genes for transfer, particularly noteworthy are those that improve the efficacy of therapy, such as by promoting the survival and / or function of transplanted cells; genes that provide genetic markers for cell selection and / or evaluation, e.g., genes for evaluating in vivo survival or localization; and genes that improve safety, e.g., by making cells susceptible to negative selection in vivo, as described in Lupton SD et al., Mol. and Cell Biol., 11:6 (1991) and Riddell et al., Human Gene Therapy 3:319-338 (1992); see also, e.g., PCT / US91 / 08442 and PCT / US94 / 05601 by Lupton et al., which describe the use of bifunctional selectable fusion genes derived from the fusion of dominant positive and negative selectable markers. See, e.g., Riddell et al., U.S. Patent No. 6,040,177, paragraphs 14-17.

[0306] Preparation of engineered cells In some embodiments, preparation of engineered cells involves one or more culture and / or preparation steps. Cells for introducing HLA-peptide-ABP, e.g., TCR, can be isolated from a biological sample, e.g., a sample obtained from or derived from a subject. In some embodiments, the subject from whom the cells are isolated has a disease or condition, or is in need of cell therapy, or is a subject to whom a cell therapy drug will be administered. In some embodiments, the subject is a human in need of a particular therapeutic intervention, such as adoptive cell therapy, in which cells are isolated, treated, and / or engineered.

[0307] Thus, in some embodiments, the cells are primary cells, e.g., primary human cells. Samples include tissues, body fluids, and other samples taken directly from a subject, as well as samples obtained by one or more processing steps, such as separation, centrifugation, genetic manipulation (e.g., transduction with a virus vector), washing, and / or incubation. Biological samples can be samples obtained directly from a biological source or processed samples. Biological samples include, but are not limited to, body fluids such as blood, plasma, serum, cerebrospinal fluid, synovial fluid, urine, and sweat, tissue and organ samples, e.g., processed samples derived therefrom.

[0308] In some embodiments, the sample from which the cells are derived or isolated is a blood or blood-derived sample, or is or is derived from an apheresis or leukapheresis product. Exemplary samples include whole blood, peripheral blood mononuclear cells (PBMCs), white blood cells, bone marrow, thymus, tissue biopsy, tumor, leukemia, lymphoma, lymph node, gut-associated lymphoid tissue, mucosa-associated lymphoid tissue, spleen, other lymphoid tissue, liver, lung, stomach, intestine, colon, kidney, pancreas, breast, bone, prostate, cervix, testis, ovary, tonsil, or other organ, and / or cells derived therefrom. Samples include autologous and allogeneic samples in the context of cell therapy, e.g., adoptive cell therapy.

[0309] In some embodiments, the cells are derived from a cell line, e.g., a T cell line. The cells are, in some embodiments, obtained from a heterologous source, e.g., mouse, rat, non-human primate, or pig.

[0310] In some embodiments, cell isolation involves one or more preparative and / or affinity-based cell separation steps. In some instances, cells are washed, centrifuged, and / or incubated in the presence of one or more reagents, e.g., to remove unwanted components, enrich for desired components, or lyse or remove cells sensitive to a particular reagent. In some instances, cells are separated based on one or more properties, such as density, adhesion properties, size, sensitivity, or resistance to a particular component.

[0311] In some examples, cells from the subject's circulating blood are obtained, for example, by apheresis or leukapheresis. The sample, in some embodiments, includes lymphocytes, e.g., T cells, monocytes, granulocytes, B cells, other nucleated leukocytes, red blood cells, and / or platelets, and in some embodiments, includes cells other than red blood cells and platelets.

[0312] In some embodiments, blood cells collected from a subject are washed, e.g., to remove the plasma fraction and place the cells in an appropriate buffer or medium in preparation for subsequent processing steps. In some embodiments, the cells are washed with phosphate-buffered saline (PBS). In some embodiments, the wash solution lacks calcium and / or magnesium and / or many or all divalent cations. In some aspects, the wash step is performed using a semi-automated "flow-through" centrifuge (e.g., a Cobe 2991 cell processor from Baxter) according to the manufacturer's instructions. In some aspects, the wash step is performed using tangential flow filtration (TFF) according to the manufacturer's instructions. In some embodiments, the cells are resuspended in various biocompatible buffers after washing, such as, for example, Ca++ / Mg++-free PBS. In certain embodiments, the components of the blood cell sample are removed and the cells are resuspended directly in culture medium.

[0313] In some embodiments, the method involves preparing white blood cells from peripheral blood by a density-based cell separation method, for example, by lysing red blood cells and centrifuging through a Percoll or Ficoll gradient.

[0314] In some embodiments, isolation methods involve separating different cell types based on the intracellular expression or presence of one or more specific molecules, e.g., surface markers, such as surface proteins, intracellular markers, or nucleic acids. In some embodiments, any known method for performing separation based on such markers can be used. In some embodiments, separation is affinity- or immunoaffinity-based separation. For example, in some aspects, isolating involves separating cells and cell populations based on the expression or expression level of one or more markers, typically cell surface markers, e.g., by incubation with an antibody or binding partner that specifically binds to such markers, followed, typically by a washing step, and separating cells bound to the antibody or binding partner from cells that are not bound to the antibody or binding partner.

[0315] Such separation steps can be based on positive selection, in which cells that bind to the reagent are retained for further use, and / or negative selection, in which cells that do not bind to the antibody or binding partner are retained. In some instances, both fractions are retained for further use. In some embodiments, negative selection is particularly useful when antibodies that specifically identify cell types in a heterogeneous population are not available, such that separation is optimally performed based on markers expressed by cells other than the desired population.

[0316] Separation does not necessarily require enrichment or removal of 100% of a particular cell population or the cells that express a particular marker.For example, positive selection or enrichment of a particular type of cell, such as the cells that express a marker, refers to increasing the number or proportion of such cells, but does not necessarily require the complete elimination of cells that do not express the marker.Similarly, negative selection, removal or depletion of a particular type of cell, such as the cells that express a marker, refers to reducing the number or proportion of such cells, but does not necessarily require the complete elimination of all such cells.

[0317] In some instances, multiple rounds of separation steps are performed, with fractions positively or negatively selected in one step being subjected to a subsequent separate separation step, e.g., positive or negative selection. In some instances, a single separation step can deplete cells that simultaneously express multiple markers, e.g., by incubating cells with multiple antibodies or binding partners, e.g., each specific for a marker targeted by negative selection. Similarly, multiple cell types can be simultaneously positively selected by incubating cells with multiple antibodies or binding partners expressed in different cell types.

[0318] For example, in some embodiments, specific subpopulations of T cells, e.g., cells that express positive or high levels of one or more surface markers, e.g., CD28+, CD62L+, CCR7+, CD27+, CD127+, CD4+, CD8+, CD45RA+, and / or CD45RO+ T cells, are isolated by positive or negative selection techniques.

[0319] For example, CD3+, CD28+ T cells can be positively selected using CD3 / CD28 conjugated magnetic beads, such as DYNABEADS® M-450 CD3 / CD28 T cell expander.

[0320] In some embodiments, isolation is achieved by enriching for a particular cell population by positive selection or depleting a particular cell population by negative selection. In some embodiments, a specific gene is expressed (marker +) or expressed at relatively high levels (marker +) on positively or negatively selected cells, respectively. 高 Positive or negative selection is achieved by incubating the cells with one or more antibodies or other binding agents that specifically bind to the surface or surfaces identified.

[0321] In some embodiments, T cells are isolated from a peripheral blood mononuclear cell (PBMC) sample by negatively selecting for markers expressed on non-T cells, e.g., B cells, monocytes, or other leukocytes, such as CD14. In some aspects, a CD4+ or CD8+ selection step is used to separate CD4+ helper T cells from CD8+ cytotoxic T cells. Such CD4+ and CD8+ populations can be further sorted into subpopulations by positively or negatively selecting for markers expressed on, or at relatively high levels of, one or more naive, memory, and / or effector T cell subpopulations.

[0322] In some embodiments, naive, central memory, effector memory, and / or central memory stem cells in CD8+ cells are further enriched or depleted, e.g., by positive or negative selection based on surface antigens associated with each subpopulation. In some embodiments, enrichment of central memory T (TCM) cells is achieved to improve efficacy, e.g., long-term survival, proliferation, and / or engraftment following administration, which in some embodiments is particularly robust for such subpopulations. See Terakura et al. (2012) Blood. 1:72-82; Wang et al. (2012) J Immunother. 35(9):689-701. In some embodiments, combining TCM-enriched CD8+ T cells with CD4+ T cells further enhances efficacy.

[0323] In embodiments, memory T cells reside in both the CD62L+ and CD62L- subsets of CD8+ peripheral blood lymphocytes. For example, anti-CD8 and anti-CD62L antibodies may be used to enrich or deplete the CD62L-CD8+ and / or CD62L+CD8+ fractions of peripheral blood mononuclear cells (PBMCs).

[0324] In some embodiments, enrichment of central memory T (TCM) cells is based on positive or high surface expression of CD45RO, CD62L, CCR7, CD28, CD3, and / or CD127, and in some aspects based on negative selection of cells expressing or highly expressing CD45RA and / or granzyme B. In some aspects, a CD8+ population enriched for TCM cells is isolated by depletion of cells expressing CD4, CD14, CD45RA, and positive selection or enrichment of cells expressing CD62L. In one aspect, enrichment of central memory T (TCM) cells is performed first from a negative fraction of cells selected based on CD4 expression, followed by negative selection based on CD14 and CD45RA expression, and positive selection based on CD62L. Such selections are performed simultaneously in some aspects and sequentially in either order in other aspects. In some embodiments, the same CD4 expression-based selection step used in preparing the CD8+ cell population or subpopulation is also used to generate a CD4+ cell population or subpopulation, e.g., both the positive and negative fractions from the CD4-based separation are retained and used in subsequent steps of the method, optionally followed by one or more additional positive or negative selection steps.

[0325] In a particular example, a sample of PBMCs or other leukocytes, retaining both negative and positive fractions, is subjected to selection of CD4+ cells. The negative fraction is then subjected to negative selection based on expression of CD14 and CD45RA or ROR1, and positive selection based on characteristic markers of central memory T cells, such as CD62L or CCR7. The positive and negative selections can be performed in any order.

[0326] CD4+ T helper cells are sorted into naive cells, central memory cells, and effector cells by identifying cell populations that possess cell surface antigens. CD4+ lymphocytes can be obtained using standard methods. In some embodiments, naive CD4+ T lymphocytes are CD45RO-, CD45RA+, CD62L+, CD4+ T cells. In some embodiments, central memory CD4+ cells are CD62L+ and CD45RO+. In some embodiments, effector CD4+ cells are CD62L- and CD45RO-.

[0327] In one example, to enrich for CD4+ cells by negative selection, a monoclonal antibody cocktail typically includes antibodies against CD14, CD20, CD11b, CD16, HLA-DR, and CD8. In some embodiments, antibodies or binding partners are attached to a solid support or substrate, such as magnetic or paramagnetic beads, to allow for cell separation following positive and / or negative selection. For example, in some embodiments, cells and cell populations are separated or isolated using immunomagnetic (or affinity magnetic) separation techniques (reviewed in Methods in Molecular Medicine, vol. 58: Metastasis Research Protocols, Vol. 2: Cell Behavior In Vitro and In Vivo, pp 17-25 Edited by: SA Brooks and U. Schumacher Humana Press Inc., Totowa, NJ).

[0328] In some embodiments, a sample or composition of cells to be separated is incubated with a small, magnetizable or magnetically responsive material, e.g., magnetically responsive particles or microparticles such as paramagnetic beads (e.g., Dynabeads or MACS beads, etc.). The magnetically responsive material, e.g., particles, are generally attached directly or indirectly to a binding partner, e.g., an antibody, that specifically binds to a molecule, e.g., a surface marker, present on the cell(s) or population of cells desired to be separated, e.g., negatively or positively selected.

[0329] In some embodiments, the magnetic particles or beads comprise a magnetically responsive material bound to a specific binding member, e.g., an antibody or other binding partner. There are many well-known magnetically responsive materials for use in magnetic separation methods. Suitable magnetic particles include those described in Molday, U.S. Pat. No. 4,452,773, and European Patent Specification EP 452342(B), each of which is incorporated by reference in its entirety. Other examples include colloidal-sized particles, such as those described in Owen, U.S. Pat. No. 4,795,698, and Liberti et al., U.S. Pat. No. 5,200,084.

[0330] Incubation is generally carried out under conditions in which molecules, such as antibodies or binding partners attached to the magnetic particles or beads, or secondary antibodies or other reagents that specifically bind to such antibodies or binding partners, specifically bind to the cell surface, if present on cells in the sample.

[0331] In some embodiments, the sample is placed in a magnetic field, and cells with attached magnetically responsive or magnetizable particles are attracted to the magnet and separated from unlabeled cells. Positive selection retains cells that are attracted to the magnet, while negative selection retains cells that are not attracted (unlabeled cells). In some embodiments, a combination of positive and negative selection is performed during the same selection step, and the positive and negative fractions are retained for further processing or further separation steps.

[0332] In certain embodiments, magne...

Claims

1. 1. An antigen binding protein (ABP) that specifically binds to an HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, wherein the HLA class I molecule and the HLA-restricted peptide are each selected from the HLA-peptide antigens set forth in any one of SEQ ID NOS: 10,755 to 29,364, and the ABP comprises a T cell receptor (TCR) or an antigen-binding fragment thereof.

2. 2. The ABP of claim 1, wherein the HLA-restricted peptide is about 5 to 15 amino acids in length.

3. 3. The ABP of claim 2, wherein the HLA-restricted peptide is about 8-12 amino acids in length, optionally 8, 9, 10, 11, or 12 amino acids in length.

4. The HLA-peptide antigen is a. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGADGVGK; b. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK; c. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; d. CTNNB1_S45P MHC class I antigen containing HLA-A*03:01 and the restriction peptide TTAPPLSGK; e. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGADGVGK; f. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGAVGVGK; g. RAS_G12V MHC class I antigen comprising HLA-C*01:02 and the restriction peptide AVGVGKSAL; h. RAS_G12V MHC class I antigen comprising HLA-A*03:01 and the restriction peptide VVVGAVGVGK; i. TP53_K132N MHC class I antigen containing HLA-A*24:02 and the restrictive peptide TYSPALNNMF; j. CTNNB1_S37Y MHC class I antigen comprising HLA-A*02:01 and the restriction peptide YLDSGIHYGA; k. RAS_G12C MHC class I antigen comprising HLA-A*03:01 and the restriction peptide VVVGACGVGK; l. RAS_G12C MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGACGVGK; m. RAS_G12D MHC class I antigen comprising HLA-A*03:01 and the restriction peptide VVVGADGVGK; n. RAS_Q61H MHC class I antigen comprising HLA-A*01:01 and the restriction peptide ILDTAGHEEY; and o. TP53_R213L MHC class I antigen containing A*02:01 and the restriction peptide YLDDRNTFL 10. The ABP of any one of the preceding claims, selected from the group consisting of:

5. a. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-A*02:01; b. the restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-A*02:06; c. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*27:05; d. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*35:01; e. the restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*41:02; f. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*48:01; g. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-C*08:03; h. The restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; i. the restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; j. The restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*03:02; k. The restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*68:01; l. The restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-B*27:05; m. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*02:01; n. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*02:05; o. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*03:01; p. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*11:01; q. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*11:01; r. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*26:01; s. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*31:01; t. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*68:01; u. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*07:02; v. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*08:01; w. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*13:02; x. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*15:01; y. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*27:05; z. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*35:01; aa. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*37:01; bb. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*38:01; cc. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*40:01; dd. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*40:02; ee. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*44:02; ff. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*44:03; gg. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*48:01; hh. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*50:01; ii. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*57:01; jj. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*01:02; kk. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*02:02; ll. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*03:03; mm. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*03:04; nn. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*04:01; oo. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*05:01; pp. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*07:04; qq. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*08:02; rr. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*08:03; ss. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*16:01; tt. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*17:01; The restricted peptide contains the RAS_G12R mutation and the HLA class I molecule is HLA-B*41:02; vv. the restricted peptide contains the RAS_G12R mutation and the HLA class I molecule is HLA-C*07:04; The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:01; xx. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:05; yy. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:06; zz. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*03:01; aaa. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*03:01; bbb. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*11:01; ccc. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*11:01; ddd. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*25:01; eee. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*26:01; fff. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*30:01; ggg. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*31:01; hhh. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*31:01; iii. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*32:01; jjj. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*68:02; kkk. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*07:02; lll. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*08:01; mmm. the restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*13:02; nnn. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*14:02; ooo. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*15:01; ppp. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*27:05; qqq. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*39:01; rrr. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*40:01; sss. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*40:02; ttt. the restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*41:02; uuu. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*44:05; vvv. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*50:01; www. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*51:01; xxx. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*01:02; yyy. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*01:02; zzz. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*03:03; aaaa. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*03:04; bbbb. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*08:02; cccc. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*14:02; dddd. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*17:01; eeee. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-A*02:01; ffff. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*07:02; The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*08:01; hhhh. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:01; iii. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:03; jjjj. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:08; kkkk. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*38:01; lllll. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-C*04:01; mmmm. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*01:01; nnnn. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*02:01; The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*23:01; pppp. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*29:01; qqqq. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*30:02; rrrr. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*33:01; ssss. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*68:01; tttt. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*07:02; uuuu. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*08:01; The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*18:01; www. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*35:01; xxxx. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*38:01; yyyy. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*40:01; zzzz. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*44:02; aaaaa. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-C*03:04; bbbbbb. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-C*05:01; or ccccc. The restricted peptide contains the KRAS_Q61H mutation, and the HLA class I molecule is HLA-C*08:02; The ABP according to any one of claims 1 to 3.

6. a. the restricted peptide contains a KRAS_G13D mutation and the HLA class I molecule is C*08:02 or A*11:01; b. The restricted peptide contains the KRAS_Q61K mutation and the HLA class I molecule is A*01:01; c. The restricted peptide contains the NRAS_Q61K mutation and the HLA class I molecule is A*01:01; d. the restricted peptide contains the TP53_R249M mutation and the HLA class I molecule is B*35:12, B*35:03, or B*35:01; e. the restricted peptide comprises the CTNNB1_S45P mutation and the HLA class I molecule is A*03:01, A*11:01, A*68:01, or A*03:02; f. the restricted peptide comprises the CTNNB1_S45F mutation and the HLA class I molecule is A*03:01, A*11:01, or A*68:01; g. The restriction peptide contains the ERBB2_Y772_A775dup mutation and the HLA class I molecule is B*18:01; h. the restricted peptide contains a KRAS_G12D mutation and the HLA class I molecule is A*11:01, A*03:01, or C*08:02; i. the restricted peptide contains the NRAS_G12D mutation and the HLA class I molecule is A*11:01, A*03:01, or C*08:02; j. The restricted peptide contains the KRAS_Q61R mutation and the HLA class I molecule is A*01:01; k. The restricted peptide contains the NRAS_Q61R mutation and the HLA class I molecule is A*01:01; l. The restricted peptide comprises the CTNNB1_T41A mutation and the HLA class I molecule is A*03:01, A*03:02, A*11:01, B*15:10, C*03:03, or C*03:04; m. the restricted peptide contains the TP53_K132N mutation and the HLA class I molecule is A*24:02 or A*23:01; n. The restricted peptide contains a KRAS_G12A mutation and the HLA class I molecule is A*03:01 or A*11:01; o. the restricted peptide contains the KRAS_Q61L mutation and the HLA class I molecule is A*01:01; p. the restricted peptide contains the NRAS_Q61L mutation and the HLA class I molecule is A*01:01; q. the restricted peptide comprises the TP53_R213L mutation and the HLA class I molecule is A*02:07, C*08:02, or A*02:01; r. The restricted peptide contains the BRAF_G466V mutation and the HLA class I molecule is B*15:01 or B*15:03; s. the restricted peptide contains a KRAS_G12V mutation and the HLA class I molecule is A*03:01, A*03:02, A*11:01, or C*01:02; t. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is A*01:01; u. The restricted peptide contains the NRAS_Q61H mutation and the HLA class I molecule is A*01:01; v. the restricted peptide comprises the CTNNB1_S37F mutation and the HLA class I molecule is A*01:01, A*23:01, A*24:02, B*15:10, B*39:06, C*05:01, C*14:02, or C*14:03; w. the restricted peptide contains the TP53_S127Y mutation and the HLA class I molecule is A*11:01 or A*03:01; x. The restricted peptide contains the TP53_K132E mutation and the HLA class I molecule is A*24:02, C*14:03, or A*23:01; y. the restricted peptide contains a KRAS_G12C mutation and the HLA class I molecule is A*02:01, A*11:01, or A*03:01; z. the restricted peptide contains the NRAS_G12C mutation and the HLA class I molecule is A*02:01, A*11:01, or A*03:01; aa. The restricted peptide contains the EGFR_L858R mutation, and the HLA class I molecule is A*11:01 or A*03:01; bb. the restricted peptide comprises the TP53_Y220C mutation and the HLA class I molecule is A*02:01; or cc. The restricted peptide contains the TP53_R175H mutation, and the HLA class I molecule is A*02:01; The ABP according to any one of claims 1 to 3.

7. The HLA-peptide antigen is a. CTNNB1_S45P MHC class I antigen comprising A*11:01 and the constrained peptidic peptide TTAPPLSGK; b. CTNNB1_T41A MHC class I antigen containing A*11:01 and the constraint peptide ATAPSLSGK; c. RAS_G12D MHC class I antigen comprising A*11:01 and the constraint peptide VVVGADGVGK; d. RAS_G12V MHC class I antigen comprising A*03:01 and the constraint peptide VVGAVGVGK; e. RAS_G12V MHC class I antigen comprising A*03:01 and the constraint peptide VVVGAVGVGK; f. RAS_G12V MHC class I antigen comprising A*11:01 and the constraint peptide VVGAVGVGK; g. RAS_G12V MHC class I antigen comprising A*11:01 and the constraint peptide VVVGAVGVGK; h. KRAS_Q61R MHC class I antigen comprising A*01:01 and the constraint peptide ILDTAGREEY; and i. TP53_R213L MHC class I antigen containing A*02:01 and the constraint peptide YLDDRNTFL The ABP according to any one of claims 1 to 3, selected from:

8. The ABP of any one of claims 1 to 3, wherein the HLA-restricted peptide comprises a RAS G12 mutation.

9. 9. The ABP of claim 8, wherein the G12 mutation is a G12C, G12D, G12V, or G12A mutation.

10. 9. The ABP of claim 8, wherein the HLA-peptide antigen comprises an HLA class I molecule selected from HLA-A*02:01, HLA-A*11:01, HLA-A*31:01, HLA-C*01:02, and HLA-A*03:

01.

11. The ABP of any one of claims 8 to 10, wherein the RAS G12 mutation is any one or more of a KRAS mutation, an NRAS mutation, and an HRAS mutation.

12. The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12C MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGACGVGK; c. RAS_G12C MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGACGVGK; d. RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK; e. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGADGVGK; f. RAS_G12D MHC class I antigen comprising HLA-A*03:01 and the restriction peptide VVVGADGVGK; g. RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGAVGVGK; h. RAS_G12V MHC class I antigen comprising HLA-A*31:01 and the restriction peptide VVVGAVGVGK; i. RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVGAVGVGK; j. RAS_G12V MHC class I antigen comprising HLA-C*01:02 and the restriction peptide AVGVGKSAL; and k. RAS_G12V MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGAVGVGK 10. The ABP of claim 9 selected from:

13. The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGADGVGK; c. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGADGVGK; d. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK; e. RAS_G12V MHC class I antigen containing HLA-A*31:01 and the restriction peptide VVVGAVGVGK; f. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGAVGVGK; g. RAS_G12V MHC class I antigen comprising HLA-C*01:02 and the restriction peptide AVGVGKSAL; and h. RAS_G12V MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGAVGVGK 10. The ABP of claim 9 selected from:

14. The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK; and c. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK 10. The ABP of claim 9 selected from:

15. 10. The ABP of claim 9, wherein the HLA-peptide antigen is a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV.

16. 10. The ABP of claim 9, wherein the HLA-peptide antigen is a RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK.

17. 10. The ABP of claim 9, wherein the HLA-peptide antigen is a RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGAVGVGK.

18. The ABP of any one of claims 1 to 3, wherein the HLA-restricted peptide comprises a RAS Q61 mutation.

19. 19. The ABP of claim 18, wherein the Q61 mutation is a Q61H, Q61K, Q61R, or Q61L mutation.

20. 19. The ABP of claim 18, wherein the HLA-peptide antigen is the RAS_Q61H MHC class I antigen comprising HLA-A*01:01 and the restriction peptide ILDTAGHEEY.

21. The ABP of any one of claims 1 to 3, wherein the HLA-restricted peptide comprises a TP53 mutation.

22. 22. The ABP of claim 21, wherein the TP53 mutation comprises an R213L, S127Y, Y220C, R175H, or R249M mutation.

23. 22. The ABP of claim 21, wherein the HLA-peptide antigen is the TP53 R213L MHC class I antigen comprising A*02:01 and the restriction peptide YLDDRNTFL.

24. 10. The ABP of any one of the preceding claims, which binds to the HLA-peptide antigen through at least one contact point with the HLA class I molecule and through at least one contact point with the HLA-restricted peptide.

25. 10. The ABP of any one of the preceding claims, wherein the antigen binding protein binds to a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV, and wherein the ABP binds to the RAS_G12C MHC class I antigen with higher affinity than an HLA-peptide antigen comprising a different RAS G12 mutation.

26. 26. The ABP of claim 25, which binds to the RAS_G12C MHC class I antigen with higher affinity than an HLA-peptide antigen comprising the constraint peptide KLVVVGAVGV and an HLA-A2 molecule.

27. 27. The ABP of claim 26, which does not bind to an HLA-peptide antigen comprising the constraint peptide KLVVVGAVGV and an HLA-A2 molecule.

28. 10. The ABP of any one of the preceding claims, wherein the antigen binding protein is linked to a scaffold, optionally the scaffold comprises serum albumin or Fc, and optionally the Fc is human Fc and is an IgG (IgG1, IgG2, IgG3, IgG4), IgA (IgA1, IgA2), IgD, IgE, or IgM isotype Fc.

29. 10. The ABP of any one of the preceding claims, wherein the antigen binding protein is linked to the scaffold via a linker, optionally the linker is a peptide linker, optionally the peptide linker is a hinge region of a human antibody.

30. 10. The ABP of any one of the preceding claims, wherein the TCR or antigen-binding portion thereof comprises a TCR variable region.

31. 10. The ABP of any one of the preceding claims, wherein the TCR or antigen-binding portion thereof comprises one or more TCR complementarity-determining regions (CDRs).

32. 10. The ABP of any one of the preceding claims, wherein the TCR comprises an alpha chain and a beta chain.

33. 10. The ABP of any one of the preceding claims, wherein the TCR comprises a gamma chain and a delta chain.

34. 10. The ABP of any one of the preceding claims, wherein the TCR comprises a single chain TCR (scTCR).

35. 10. The ABP of any one of the preceding claims, wherein the TCR comprises a recombinant TCR sequence.

36. 10. The ABP of any one of the preceding claims, wherein the TCR comprises a human TCR sequence, and optionally the human TCR sequence is a fully human TCR sequence.

37. 10. The ABP of any one of the preceding claims, wherein the TCR comprises a modified TCR alpha constant (TRAC) region, a modified TCR beta constant (TRBC) region, or a modified TRAC region and a modified TRBC region.

38. 10. The ABP of any one of the preceding claims, comprising a modification that extends its half-life.

39. 10. The ABP of any one of the preceding claims, which is part of a chimeric antigen receptor (CAR) comprising an extracellular portion comprising an antigen binding protein and an intracellular signaling domain.

40. 40. The ABP of claim 39, wherein the intracellular signaling domain comprises an ITAM.

41. 41. The ABP of claim 39 or 40, wherein the intracellular signaling domain comprises the signaling domain of the zeta chain of the CD3-zeta (CD3) chain.

42. 42. The antigen binding protein of any one of claims 39 to 41, further comprising a transmembrane domain linking said extracellular domain and said intracellular signaling domain.

43. 43. The ABP of claim 42, wherein the transmembrane domain comprises the transmembrane portion of CD28.

44. 44. The antigen-binding protein of any one of claims 39 to 43, further comprising an intracellular signalling domain of a T cell co-stimulatory molecule.

45. 45. The ABP of claim 44, wherein the T cell costimulatory molecule is CD28, 4-1BB, OX-40, ICOS, or any combination thereof.

46. 10. An ABP according to any one of the preceding claims for use as a medicament.

47. 10. The ABP of any one of the preceding claims for use in treating cancer, optionally wherein said cancer expresses or is predicted to express said HLA-peptide antigen.

48. 10. The ABP of any one of the preceding claims for use in the treatment of cancer, wherein the cancer is selected from solid tumors and hematological tumors.

49. 10. An antigen binding protein (ABP) that competes for binding with the ABP of any one of the preceding claims.

50. 10. An antigen binding protein (ABP) that binds to the same HLA-peptide antigen epitope as that bound by the ABP of any one of the preceding claims.

51. 10. An engineered cell expressing a receptor comprising an antigen binding protein according to any one of the preceding claims.

52. 52. The engineered cell of claim 51, which is a T cell.

53. 53. The engineered cell of claim 52, wherein the T cells are selected from the group consisting of naive T (TN) cells, effector T cells (TEFF), memory T cells, stem cell memory T cells (TSCM), central memory T cells (TCM), effector memory T cells (TEM), terminally differentiated effector memory T cells, tumor infiltrating lymphocytes (TIL), immature T cells, mature T cells, helper T cells, cytotoxic T cells, mucosal-associated invariant T (MALT) cells, regulatory T cells (Treg), TH1 cells, TH2 cells, TH3 cells, TH17 cells, TH9 cells, TH22 cells, follicular helper T cells, natural killer T cells (NKT), alpha-beta T cells, and gamma-delta T cells.

54. 53. The engineered cell of claim 52, wherein the T cell is a cytotoxic T cell (CTL).

55. 55. The engineered cell of any one of claims 51 to 54, which is a human cell or a human-derived cell.

56. 56. The engineered cell of any one of claims 51 to 55, which is an autologous cell of the subject.

57. 57. The engineered cell of claim 56, wherein the subject is known to have or suspected of having cancer.

58. 58. The engineered cell of any one of claims 56-57, wherein the autologous cell is a cell isolated from the subject.

59. 59. The engineered cell of claim 58, wherein the isolated cells are ex vivo cultured cells, and optionally the in vivo cultured cells are stimulated cells.

60. 58. The engineered cell of any one of claims 56 to 57, wherein the autologous cell is an in vivo engineered cell.

61. 61. The engineered cell of any one of claims 51 to 60, wherein the antigen binding protein is expressed by a heterologous promoter.

62. 62. The engineered cell of any one of Claims 51-61, wherein the ABP comprises a T cell receptor (TCR) or an antigen-binding portion thereof, and wherein a polynucleotide encoding the T cell receptor (TCR) or an antigen-binding portion thereof is inserted into an endogenous TCR locus.

63. 63. The engineered cell of any one of claims 51 to 62, which does not express an endogenous ABP.

64. 10. An isolated polynucleotide encoding an ABP or antigen-binding portion thereof according to any one of the preceding claims, or a set of polynucleotides encoding an ABP or antigen-binding portion thereof according to any one of the preceding claims.

65. 65. A vector or set of vectors comprising the polynucleotide or set of polynucleotides of claim 64.

66. 65. A virus comprising the isolated polynucleotide or set of polynucleotides of claim 64.

67. 67. The virus of claim 66, which is a filamentous phage.

68. A yeast cell comprising an isolated polynucleotide or set of polynucleotides according to any one of the preceding claims.

69. 66. A host cell comprising a polynucleotide or set of polynucleotides according to any one of the preceding claims or a vector or set of vectors according to claim 65, optionally being a CHO or HEK293 or optionally being a T cell.

70. 70. A method of producing an antigen binding protein, comprising expressing said antigen binding protein using the host cell of claim 69 and isolating the expressed antigen binding protein.

71. 10. A pharmaceutical composition comprising an antigen-binding protein according to any one of the preceding claims and a pharmaceutically acceptable excipient.

72. 10. A method of treating cancer in a subject, comprising administering to the subject the ABP of any one of the preceding claims, the engineered cell of any one of claims 51-63, or the pharmaceutical composition of claim 71, optionally wherein the cancer is selected from a solid tumor and a hematological tumor.

73. 10. A method of stimulating an immune response in a subject, comprising administering to the subject the ABP of any one of the preceding claims, the engineered cell of any one of claims 51-63, or the pharmaceutical composition of claim 71, optionally wherein the subject has cancer, and optionally wherein the cancer is selected from a solid tumor and a hematological tumor.

74. 10. A method of killing target cells in a subject, comprising administering to the subject an ABP of any one of the preceding claims, an engineered cell of any one of claims 51-63, or a pharmaceutical composition of claim 71, optionally wherein the subject has cancer and the target cells are cancer cells, optionally wherein the cancer is selected from a solid tumor and a hematological tumor.

75. 75. The method of any one of claims 72 to 74, wherein the subject is a human subject.

76. 75. The method of any one of claims 72 to 74, wherein the cancer expresses or is predicted to express an HLA-peptide antigen or an HLA class I molecule set forth in any one of SEQ ID NOs: 10,755 to 29,364, and the ABP binds to the HLA-peptide antigen.

77. 75. The method of any one of claims 72 to 74, wherein the cancer expresses or is predicted to express an HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, the HLA-restricted peptide being located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, wherein the HLA class I molecule and the HLA-restricted peptide are each selected from the HLA-peptide antigens set forth in any one of SEQ ID NOs: 10,755 to 29,364, and wherein the ABP binds to the HLA-peptide antigen.

78. The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGADGVGK; c. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK; d. CTNNB1_S45P MHC class I antigen containing HLA-A*03:01 and the restriction peptide TTAPPLSGK; e. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGADGVGK; f. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGAVGVGK; g. RAS_G12V MHC class I antigen comprising HLA-C*01:02 and the restriction peptide AVGVGKSAL; h. RAS_G12V MHC class I antigen comprising HLA-A*03:01 and the restriction peptide VVVGAVGVGK; i. TP53_K132N MHC class I antigen containing HLA-A*24:02 and the restrictive peptide TYSPALNNMF; j. CTNNB1_S37Y MHC class I antigen comprising HLA-A*02:01 and the restriction peptide YLDSGIHYGA; k. RAS_G12C MHC class I antigen comprising HLA-A*03:01 and the restriction peptide VVVGACGVGK; l. RAS_G12C MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGACGVGK; m. RAS_G12D MHC class I antigen comprising HLA-A*03:01 and the restriction peptide VVVGADGVGK; n. RAS_Q61H MHC class I antigen comprising HLA-A*01:01 and the restriction peptide ILDTAGHEEY; and o. TP53_R213L MHC class I antigen containing A*02:01 and the restriction peptide YLDDRNTFL 78. The method of claim 77, wherein the

79. a. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-A*02:01; b. the restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-A*02:06; c. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*27:05; d. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*35:01; e. the restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*41:02; f. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*48:01; g. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-C*08:03; h. The restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; i. the restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; j. The restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*03:01; k. The restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*03:02; l. The restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*68:01; m. The restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-B*27:05; n. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*02:01; o. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*02:05; p. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*03:01; q. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*11:01; r. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*11:01; s. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*26:01; t. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*31:01; u. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*68:01; v. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*07:02; w. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*08:01; x. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*13:02; y. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*15:01; z. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*27:05; aa. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*35:01; bb. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*37:01; cc. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*38:01; dd. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*40:01; ee. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*40:02; ff. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*44:02; gg. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*44:03; hh. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*48:01; ii. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*50:01; jj. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*57:01; kk. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*01:02; 11. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*02:02; mm. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*03:03; nn. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*03:04; oo. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*04:01; pp. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*05:01; qq. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*07:04; rr. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*08:02; ss. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*08:03; tt. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*16:01; The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*17:01; vv. The restricted peptide contains the RAS_G12R mutation and the HLA class I molecule is HLA-B*41:02; The restricted peptide contains the RAS_G12R mutation and the HLA class I molecule is HLA-C*07:04; xx. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:01; yy. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:05; zz. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:06; aaa. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*03:01; bbb. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*03:01; ccc. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*11:01; ddd. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*11:01; eee. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*25:01; fff. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*26:01; ggg. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*30:01; hhh. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*31:01; iii. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*31:01; jjj. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*32:01; kkk. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*68:02; lll. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*07:02; mmm. the restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*08:01; nnn. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*13:02; ooo. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*14:02; ppp. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*15:01; qqq. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*27:05; rrr. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*39:01; sss. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*40:01; ttt. the restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*40:02; The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*41:02; vvv. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*44:05; www. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*50:01; xxx. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*51:01; yyy. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*01:02; zzz. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*01:02; aaaa. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*03:03; bbbb. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*03:04; cccc. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*08:02; dddd. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*14:02; eeee. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*17:01; ffff. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-A*02:01; The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*07:02; hhhh. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*08:01; iii. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:01; jjjj. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:03; kkkk. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:08; lllll. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*38:01; mmmm. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-C*04:01; nnnn. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*01:01; oooo. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*02:01; pppp. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*23:01; qqqq. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*29:01; rrrr. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*30:02; ssss. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*33:01; tttt. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*68:01; uuuu. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*07:02; The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*08:01; www. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*18:01; xxxx. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*35:01; yyyy. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*38:01; zzzz. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*40:01; aaaaa. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*44:02; bbbbbb. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-C*03:04; cccccc. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-C*05:01; or dddddd. The restricted peptide contains the KRAS_Q61H mutation, and the HLA class I molecule is HLA-C*08:02; 78. The method of claim 77.

80. a. the restricted peptide contains a KRAS_G13D mutation and the HLA class I molecule is C*08:02 or A*11:01; b. The restricted peptide contains the KRAS_Q61K mutation and the HLA class I molecule is A*01:01; c. The restricted peptide contains the NRAS_Q61K mutation and the HLA class I molecule is A*01:01; d. the restricted peptide contains the TP53_R249M mutation and the HLA class I molecule is B*35:12, B*35:03, or B*35:01; e. the restricted peptide comprises the CTNNB1_S45P mutation and the HLA class I molecule is A*03:01, A*11:01, A*68:01, or A*03:02; f. the restricted peptide comprises the CTNNB1_S45F mutation and the HLA class I molecule is A*03:01, A*11:01, or A*68:01; g. The restriction peptide contains the ERBB2_Y772_A775dup mutation and the HLA class I molecule is B*18:01; h. the restricted peptide contains a KRAS_G12D mutation and the HLA class I molecule is A*11:01, A*03:01, or C*08:02; i. the restricted peptide contains the NRAS_G12D mutation and the HLA class I molecule is A*11:01, A*03:01, or C*08:02; j. The restricted peptide contains the KRAS_Q61R mutation and the HLA class I molecule is A*01:01; k. The restricted peptide contains the NRAS_Q61R mutation and the HLA class I molecule is A*01:01; l. the restricted peptide comprises the CTNNB1_T41A mutation and the HLA class I molecule is A*03:01, A*0302, A*11:01, B*15:10, C*03:03, or C*03:04; m. the restricted peptide contains the TP53_K132N mutation and the HLA class I molecule is A*24:02 or A*23:01; n. The restricted peptide contains a KRAS_G12A mutation and the HLA class I molecule is A*03:01 or A*11:01; o. the restricted peptide contains the KRAS_Q61L mutation and the HLA class I molecule is A*01:01; p. the restricted peptide contains the NRAS_Q61L mutation and the HLA class I molecule is A*01:01; q. the restricted peptide comprises the TP53_R213L mutation and the HLA class I molecule is A*02:07, C*08:02, or A*02:01; r. The restricted peptide contains the BRAF_G466V mutation and the HLA class I molecule is B*15:01 or B*15:03; s. the restricted peptide contains a KRAS_G12V mutation and the HLA class I molecule is A*03:01, A*03:02, A*11:01, or C*01:02; t. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is A*01:01; u. The restricted peptide contains the NRAS_Q61H mutation and the HLA class I molecule is A*01:01; v. the restricted peptide comprises the CTNNB1_S37F mutation and the HLA class I molecule is A*01:01, A*23:01, A*24:02, B*15:10, B*39:06, C*05:01, C*14:02, or C*14:03; w. the restricted peptide contains the TP53_S127Y mutation and the HLA class I molecule is A*11:01 or A*03:01; x. The restricted peptide contains the TP53_K132E mutation and the HLA class I molecule is A*24:02, C*14:03, or A*23:01; y. the restricted peptide contains a KRAS_G12C mutation and the HLA class I molecule is A*02:01, A*11:01, or A*03:01; z. the restricted peptide contains the NRAS_G12C mutation and the HLA class I molecule is A*02:01, A*11:01, or A*03:01; aa. The restricted peptide contains the EGFR_L858R mutation, and the HLA class I molecule is A*11:01 or A*03:01; bb. the restricted peptide comprises the TP53_Y220C mutation and the HLA class I molecule is A*02:01; or cc. The restricted peptide contains the TP53_R175H mutation, and the HLA class I molecule is A*02:01; 78. The method of claim 77.

81. The HLA-peptide antigen is a. CTNNB1_S45P MHC class I antigen comprising A*11:01 and the constrained peptidic peptide TTAPPLSGK; b. CTNNB1_T41A MHC class I antigen containing A*11:01 and the constraint peptide ATAPSLSGK; c. RAS_G12D MHC class I antigen comprising A*11:01 and the constraint peptide VVVGADGVGK; d. RAS_G12V MHC class I antigen comprising A*03:01 and the constraint peptide VVGAVGVGK; e. RAS_G12V MHC class I antigen comprising A*03:01 and the constraint peptide VVVGAVGVGK; f. RAS_G12V MHC class I antigen comprising A*11:01 and the constraint peptide VVGAVGVGK; g. RAS_G12V MHC class I antigen comprising A*11:01 and the constraint peptide VVVGAVGVGK; h. KRAS_Q61R MHC class I antigen comprising A*01:01 and the constraint peptide ILDTAGREEY; and i. TP53_R213L MHC class I antigen containing A*02:01 and the constraint peptide YLDDRNTFL 78. The method of claim 77, wherein the

82. 78. The method of claim 77, wherein the HLA-peptide antigen comprises an HLA-restricted peptide that is a peptide fragment of RAS containing the RAS G12 mutation.

83. 83. The method of claim 82, wherein the G12 mutation is a G12C, G12D, G12V, or G12A mutation.

84. 83. The method of claim 82, wherein the HLA-peptide antigen comprises an HLA class I molecule selected from HLA-A*02:01, HLA-A*11:01, HLA-A*31:01, HLA-C*01:02, and HLA-A*03:

01.

85. 85. The method of any one of claims 82 to 84, wherein the RAS G12 mutation is any one or more of a KRAS mutation, an NRAS mutation, and an HRAS mutation.

86. The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12C MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGACGVGK; c. RAS_G12C MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGACGVGK; d. RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK; e. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGADGVGK; f. RAS_G12D MHC class I antigen comprising HLA-A*03:01 and the restriction peptide VVVGADGVGK; g. RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGAVGVGK; h. RAS_G12V MHC class I antigen comprising HLA-A*31:01 and the restriction peptide VVVGAVGVGK; i. RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVGAVGVGK; j. RAS_G12V MHC class I antigen comprising HLA-C*01:02 and the restriction peptide AVGVGKSAL; and k. RAS_G12V MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGAVGVGK 84. The method of claim 83, wherein the

87. The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGADGVGK; c. RAS_G12D MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGADGVGK; d. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK; e. RAS_G12V MHC class I antigen containing HLA-A*31:01 and the restriction peptide VVVGAVGVGK; f. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVGAVGVGK; g. RAS_G12V MHC class I antigen comprising HLA-C*01:02 and the restriction peptide AVGVGKSAL; and h. RAS_G12V MHC class I antigen containing HLA-A*03:01 and the restriction peptide VVVGAVGVGK 84. The method of claim 83, wherein the

88. The HLA-peptide antigen is a. RAS_G12C MHC class I antigen containing HLA-A*02:01 and the restriction peptide KLVVVGACGV; b. RAS_G12D MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVVGADGVGK; or c. RAS_G12V MHC class I antigen containing HLA-A*11:01 and the restriction peptide VVVGAVGVGK 84. The method of claim 83, wherein the

89. 84. The method of claim 83, wherein the antigen binding protein binds to a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV, and the ABP binds to the RAS_G12C MHC class I antigen with higher affinity than an HLA-peptide antigen comprising a different RAS G12 mutation.

90. 90. The method of claim 89, wherein the ABP binds to the RAS_G12C MHC class I antigen with higher affinity than an HLA-peptide antigen comprising the constraint peptide KLVVVGAVGV and an HLA-A2 molecule.

91. 90. The method of claim 89, wherein the ABP does not bind to an HLA-peptide antigen comprising the constraint peptide KLVVVGAVGV and an HLA-A2 molecule.

92. 78. The method of claim 77, wherein the HLA-peptide antigen comprises an HLA-restricted peptide that is a peptide fragment of RAS containing the RAS Q61 mutation.

93. 93. The method of claim 92, wherein the Q61 mutation is a Q61H, Q61K, Q61R, or Q61L mutation.

94. 93. The method of claim 92, wherein the HLA-peptide antigen is the RAS_Q61H MHC class I antigen comprising HLA-A*01:01 and the restriction peptide ILDTAGHEEY.

95. 78. The method of claim 77, wherein the HLA-peptide antigen comprises an HLA-restricted peptide that is a peptide fragment of TP53 that contains a TP53 mutation.

96. 96. The method of claim 95, wherein the TP53 mutation comprises an R213L, S127Y, Y220C, R175H, or R249M mutation.

97. 96. The method of claim 95, wherein the HLA-peptide antigen is the TP53 R213L MHC class I antigen comprising A*02:01 and the restriction peptide YLDDRNTFL.

98. 98. The method of any one of claims 72 to 97, comprising determining or having determined the presence of any one or more of the HLA-peptide antigen, a peptide of the HLA-peptide antigen, a somatic mutation associated with the HLA-peptide antigen, and an HLA molecule of the HLA-peptide antigen in a biological sample obtained from the subject prior to said administering.

99. 99. The method of claim 98, wherein the biological sample is a blood sample or a tumor sample.

100. 100. The method of claim 99, wherein the blood sample is a plasma or serum sample.

101. 99. The method of claim 98, wherein said determining comprises RNASeq, microarray, PCR, nanostring, in situ hybridization (ISH), mass spectrometry, sequencing, or immunohistochemistry (IHC).

102. 99. The method of claim 98, wherein after determining the presence of the HLA-peptide antigen, peptide, or HLA in the biological sample obtained from the subject, an ABP that selectively binds to the HLA-peptide antigen is administered to the subject.

103. 72. A kit comprising an antigen binding protein according to any one of the preceding claims or a pharmaceutical composition according to claim 71 and instructions for use.

104. an isolated HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, and the HLA-peptide antigen is selected from the HLA-peptide antigens set forth in any one of SEQ ID NOs: 10,755 to 29,364; b. Phage display libraries and Including, the system.

105. 105. The system of claim 104, wherein the HLA-peptide antigen is bound to a solid support.

106. 106. The system of claim 105, wherein the solid support comprises a bead, a well, a membrane, a tube, a column, a plate, sepharose, a magnetic bead, a cell, or a chip.

107. 107. The system of claim 105 or 106, wherein the HLA-peptide antigen comprises a first member of an affinity binding pair and the solid support comprises a second member of the affinity binding pair.

108. 108. The system of claim 107, wherein the first member is streptavidin and the second member is biotin.

109. The system of any one of claims 104 to 108, wherein the phage display library is a human library.

110. The system of any one of claims 104 to 108, wherein the phage display library is a humanized library.

111. The system of any one of claims 104 to 110, further comprising a negative control HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, and the negative control HLA-peptide antigen comprises a different restricted peptide, a different HLA class I molecule, or a different restricted peptide and a different HLA class I molecule.

112. 112. The system of claim 111, wherein the negative control HLA-peptide antigen comprises a different restricted peptide but the same HLA class I molecule as the HLA-peptide antigen.

113. The system of any one of claims 104 to 112, comprising a reaction mixture, said reaction mixture comprising said HLA-peptide antigen and a plurality of phages from said phage display library.

114. 114. Use of the system of any one of claims 104 to 113 for identifying antigen binding proteins that selectively bind to the isolated HLA-peptide antigen.

115. A composition comprising an HLA-peptide antigen set forth by any one of SEQ ID NOs: 10,755-29,364, wherein said HLA-peptide antigen is covalently linked to an affinity tag.

116. 116. The composition of claim 115, wherein the affinity tag is a biotin tag.

117. A composition comprising an HLA-peptide antigen set forth by any one of SEQ ID NOs: 10,755-29,364 complexed to a detectable label.

118. The detectable label is ２ - The composition of claim 117, comprising a microglobulin-binding molecule.

119. Said β ２ The composition of claim 118, wherein the microglobulin-binding molecule is a labeled antibody.

120. The composition of claim 119, wherein the labeled antibody is a fluorescent dye-labeled antibody.

121. A composition comprising an HLA-peptide antigen, wherein said HLA-peptide antigen is set forth by any one of SEQ ID NOs: 10,755-29,364, bound to a solid support.

122. 122. The composition of claim 121, wherein the solid support comprises a bead, a well, a membrane, a tube, a column, a plate, sepharose, a magnetic bead, a cell, or a chip.

123. 123. The composition of claim 121 or 122, wherein the HLA-peptide antigen comprises a first member of an affinity binding pair and the solid support comprises a second member of the affinity binding pair.

124. 124. The composition of claim 123, wherein the first member is streptavidin and the second member is biotin.

125. A host cell comprising a xenogeneic HLA-peptide antigen as set forth by any one of SEQ ID NOs: 10,755-29,364.

126. A host cell expressing an HLA subtype defined by any one of the HLA-peptide antigens set forth in SEQ ID NOs: 10,755 to 29,364.

127. A host cell comprising a polynucleotide encoding an HLA-restricted peptide defined by any one of the HLA-peptide antigens in SEQ ID NOs: 10,755-29,364.

128. The host cell of claim 127, which does not contain endogenous MHC.

129. The host cell of claim 128, comprising an exogenous HLA.

130. 130. The host cell of claim 129, which is a K562 or A375 cell.

131. The host cell according to any one of claims 125 to 130, which is a cultured cell derived from a tumor cell line.

132. The host cell of claim 131, wherein the tumor cell line expresses an HLA subtype defined by the same HLA-peptide antigen as described for the HLA-restricted peptide of claim 127.

133. 132. The host cell of claim 131, wherein the tumor cell line is selected from the group consisting of HCC-1599, NCI-H510A, A375, LN229, NCI-H358, ZR-75-1, MS751, OE19, MOR, BV173, MCF-7, NCI-H82, Colo829, SK-MEL-28, KYSE270, 59M, and NCI-H146.

134. a. a host cell according to any one of claims 125 to 133; b. Cell culture medium; A cell culture system comprising:

135. 135. The cell culture system of claim 134, wherein the host cells express an HLA subtype defined by any one of the HLA-peptide antigens in SEQ ID NOs: 10,755-21,015 and SEQ ID NOs: 21,016-29,364, and the cell culture medium comprises a restricted peptide defined by the same HLA-peptide antigen as the HLA subtype.

136. 135. The cell culture system of claim 134, wherein the host cells are K562 cells comprising an exogenous HLA, the exogenous HLA being an HLA subtype defined by any one of the HLA-peptide antigens in SEQ ID NOs: 10,755-29,364, and the cell culture medium comprises a restricted peptide defined by the same HLA-peptide antigen that defines the HLA subtype.

137. 10. A method of identifying an antigen binding protein comprising providing at least one HLA-peptide antigen as set forth in SEQ ID NOs: 10,755-29,364; and binding at least one target with an antigen binding protein according to any one of the preceding claims, thereby identifying said antigen binding protein.

138. 138. The method of claim 137, wherein the antigen binding protein is present in a phage display library comprising a plurality of distinct antigen binding proteins.

139. 139. The method of claim 138, wherein the phage display library is substantially free of antigen binding proteins that non-specifically bind to the HLA of the HLA-peptide antigen.

140. 140. The method of any one of claims 137 to 139, wherein the combining step is performed two or more times, optionally at least three times.

141. 141. The method of any one of claims 137-140, further comprising contacting said antigen binding protein with one or more peptide-HLA complexes distinct from said HLA-peptide antigen to determine whether said antigen binding protein selectively binds to said HLA-peptide antigen, optionally wherein selectivity is determined by measuring the binding affinity of said antigen binding protein to a soluble target HLA-peptide complex and to a soluble HLA-peptide complex distinct from the target complex, optionally wherein selectivity is determined by measuring the binding affinity of said antigen binding protein to a target HLA-peptide complex expressed on the surface of one or more cells and to a HLA-peptide complex distinct from the target complex expressed on the surface of one or more cells.

142. 10. A method of identifying an antigen binding protein comprising obtaining at least one HLA-peptide antigen as set forth in SEQ ID NOs: 10,755-29,364; administering said HLA-peptide antigen to a subject, optionally in combination with an adjuvant; and isolating an antigen binding protein according to any one of the preceding claims from said subject.

143. 143. The method of claim 142, wherein isolating the antigen binding protein comprises screening the serum of the subject to identify the antigen binding protein.

144. 143. The method of claim 142, further comprising contacting said antigen binding protein with one or more peptide-HLA complexes distinct from said HLA-peptide antigen to determine whether said antigen binding protein selectively binds to said HLA-peptide antigen, optionally wherein selectivity is determined by measuring the binding affinity of said antigen binding protein to said HLA-peptide antigen and to a soluble HLA-peptide complex distinct from said HLA-peptide antigen, optionally wherein selectivity is determined by measuring the binding affinity of said antigen binding protein to said HLA-peptide antigen expressed on the surface of one or more cells and to a HLA-peptide complex distinct from said HLA-peptide antigen expressed on the surface of one or more cells.

145. 143. The method of claim 142, wherein the subject is a mouse, rabbit, or llama.

146. 143. The method of claim 142, wherein isolating said antigen binding protein comprises isolating B cells from said subject which express said antigen binding protein, and optionally directly cloning sequences encoding said antigen binding protein from said isolated B cells.

147. 147. The method of claim 146, further comprising using the B cells to produce hybridomas.

148. 147. The method of claim 146, further comprising cloning CDRs from the B cells.

149. 147. The method of claim 146, further comprising immortalizing the B cells, optionally via EBV transformation.

150. 147. The method of claim 146, further comprising generating a library comprising said antigen binding proteins of said B cells, optionally wherein said library is phage display or yeast display.

151. 143. The method of claim 142, further comprising humanizing the antigen binding protein.

152. 10. A method of identifying an antigen binding protein according to any one of the preceding claims, comprising obtaining a cell comprising said antigen binding protein; contacting said cell with an HLA multimer comprising at least one HLA-peptide antigen as set forth in SEQ ID NOs: 10,755-29,364; and identifying said antigen binding protein via binding between said HLA multimer and said antigen binding protein.

153. 153. The method of claim 152, further comprising contacting said cell comprising said antigen binding protein with an HLA multimer comprising a corresponding wild-type sequence of said at least one HLA-peptide antigen as set forth in SEQ ID NOs: 10,755-29,364, and excluding said antigen binding protein if said antigen binding protein binds to said HLA multimer comprising said corresponding wild-type sequence.

154. 10. A method of identifying an antigen binding protein comprising providing at least one HLA-peptide antigen as set forth in SEQ ID NOs: 10,755-29,364; and using the target to identify an antigen binding protein according to any one of the preceding claims.

155. 1. An antigen binding protein (ABP) that specifically binds to an HLA-peptide antigen comprising an HLA-restricted peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimeric portion of the HLA class I molecule, and wherein the HLA class I molecule and the HLA-restricted peptide are each selected from the HLA-peptide antigens set forth in any one of SEQ ID NOS: 10,755-29,364, and the ABP comprises an alpha CDR3 amino acid sequence and a corresponding beta CDR3 amino acid sequence selected from the group consisting of the sequences set forth in Tables 1C.1, 1C.2, 1C.3, and 1D.

156. The ABP of claim 155, further comprising an alpha variable ("V") segment, an alpha joining ("J") segment, a beta variable ("V") segment, a beta joining ("J") segment, optionally a beta diversity ("D") segment, and optionally a beta constant region selected from the group consisting of the regions set forth in Tables 1C.1, 1C.2, 1C.3, and 1D corresponding to said alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence.

157. 157. The ABP of any one of claims 155 or 156, comprising an alpha variable region and a corresponding beta variable region comprising an amino acid sequence selected from the sequences set forth in Tables 1A.1, 1A.2, 1A.3, and 1B corresponding to the alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence.

158. 1. An antigen binding protein (ABP) that specifically binds to an HLA-peptide antigen comprising an HLA-restricted RAS peptide complexed with an HLA class I molecule, wherein the HLA-restricted peptide is located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, and the HLA-restricted RAS peptide comprises at least one alteration that renders the HLA-restricted RAS peptide sequence different from the corresponding peptide sequence of a wild-type RAS peptide, and wherein the ABP comprises an alpha CDR3 amino acid sequence and a corresponding beta CDR3 amino acid sequence selected from the group consisting of the sequences shown in Tables 1C.1, 1C.2, 1C.3, and 1D.

159. a. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-A*02:01; b. the restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-A*02:06; c. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*27:05; d. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*35:01; e. the restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*41:02; f. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-B*48:01; g. The restricted peptide contains the RAS_G12A mutation and the HLA class I molecule is HLA-C*08:03; h. The restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; i. the restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*02:01; j. The restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*03:02; k. The restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-A*68:01; l. The restricted peptide contains the RAS_G12C mutation and the HLA class I molecule is HLA-B*27:05; m. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*02:01; n. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*02:05; o. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*03:01; p. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*11:01; q. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*11:01; r. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*26:01; s. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*31:01; t. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-A*68:01; u. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*07:02; v. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*08:01; w. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*13:02; x. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*15:01; y. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*27:05; z. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*35:01; aa. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*37:01; bb. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*38:01; cc. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*40:01; dd. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*40:02; ee. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*44:02; ff. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*44:03; gg. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*48:01; hh. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*50:01; ii. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-B*57:01; jj. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*01:02; kk. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*02:02; ll. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*03:03; mm. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*03:04; nn. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*04:01; oo. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*05:01; pp. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*07:04; qq. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*08:02; rr. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*08:03; ss. The restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*16:01; tt. the restricted peptide contains the RAS_G12D mutation and the HLA class I molecule is HLA-C*17:01; The restricted peptide contains the RAS_G12R mutation and the HLA class I molecule is HLA-B*41:02; vv. the restricted peptide contains the RAS_G12R mutation and the HLA class I molecule is HLA-C*07:04; The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:01; xx. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:05; yy. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*02:06; zz. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*03:01; aaa. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*03:01; bbb. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*11:01; ccc. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*11:01; ddd. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*25:01; eee. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*26:01; fff. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*30:01; ggg. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*31:01; hhh. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*31:01; iii. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*32:01; jjj. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-A*68:02; kkk. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*07:02; lll. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*08:01; mmm. the restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*13:02; nnn. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*14:02; ooo. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*15:01; ppp. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*27:05; qqq. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*39:01; rrr. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*40:01; sss. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*40:02; ttt. the restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*41:02; uuu. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*44:05; vvv. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*50:01; www. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-B*51:01; xxx. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*01:02; yyy. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*01:02; zzz. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*03:03; aaaa. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*03:04; bbbb. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*08:02; cccc. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*14:02; dddd. The restricted peptide contains the RAS_G12V mutation and the HLA class I molecule is HLA-C*17:01; eeee. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-A*02:01; ffff. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*07:02; The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*08:01; hhhh. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:01; iii. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:03; jjjj. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*35:08; kkkk. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-B*38:01; lllll. The restricted peptide contains the KRAS_G13D mutation and the HLA class I molecule is HLA-C*04:01; mmmm. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*01:01; nnnn. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*02:01; The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*23:01; pppp. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*29:01; qqqq. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*30:02; rrrr. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*33:01; ssss. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-A*68:01; tttt. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*07:02; uuuu. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*08:01; The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*18:01; www. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*35:01; xxxx. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*38:01; yyyy. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*40:01; zzzz. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-B*44:02; aaaaa. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-C*03:04; bbbbbb. The restricted peptide contains the KRAS_Q61H mutation and the HLA class I molecule is HLA-C*05:01; or ccccc. The restricted peptide contains the KRAS_Q61H mutation, and the HLA class I molecule is HLA-C*08:02; The ABP described in claim 158.

160. 160. The ABP of claim 158 or 159, wherein the HLA-peptide antigen is a RAS_G12C MHC class I antigen comprising HLA-A*02:01 and the restriction peptide KLVVVGACGV.

161. 161. The ABP of claim 160, comprising an alpha CDR3 amino acid sequence and a corresponding beta CDR3 amino acid sequence selected from the group consisting of the sequences set forth in Table 1C.

2.

162. 162. The ABP of claim 161, further comprising an alpha variable ("V") segment, an alpha joining ("J") segment, a beta variable ("V") segment, a beta joining ("J") segment, optionally a beta diversity ("D") segment, and optionally a beta constant region selected from the group consisting of the regions set forth in Table 1C.2 that correspond to said alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence.

163. 163. The ABP of claim 161 or 162, comprising an alpha variable region and a corresponding beta variable region comprising an amino acid sequence selected from the sequences set forth in Table 1A.2 corresponding to the alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence.

164. 160. The ABP of claim 158 or 159, wherein the HLA-peptide antigen is a RAS_G12V MHC class I antigen comprising HLA-A*11:01 and the restriction peptide VVGAVGVGK.

165. 165. The ABP of claim 164, comprising an alpha CDR3 amino acid sequence and a corresponding beta CDR3 amino acid sequence selected from the group consisting of the sequences set forth in Table 1C.

3.

166. 166. The ABP of claim 165, further comprising an alpha variable ("V") segment, an alpha joining ("J") segment, a beta variable ("V") segment, a beta joining ("J") segment, optionally a beta diversity ("D") segment, and optionally a beta constant region selected from the group consisting of the regions set forth in Table 1C.3 that correspond to said alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence.

167. 167. The ABP of claim 165 or 166, comprising an alpha variable region and a corresponding beta variable region comprising an amino acid sequence selected from the sequences set forth in Table 1A.3, which correspond to the alpha CDR3 amino acid sequence and the corresponding beta CDR3 amino acid sequence.

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