Antigen-binding proteins that target common antigens

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By developing antigen-binding proteins that can bind to specific HLA subtypes, the problem of difficulty in targeting and recognizing HLA molecules and peptide complexes in existing technologies has been solved, enabling highly efficient and specific recognition and attack on tumor cells.

CN112739375BActive Publication Date: 2025-10-28GRITSTONE BIO INC

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Patent Information

Application Number: CN201980060989.6
Authority / Receiving Office: CN · China
Patent Type: Patents(China)
Current Assignee / Owner
Priority Date: 2019-07-02
Filing Date: 2019-08-16
Publication Date: 2025-10-28
Estimated Expiration: 2039-08-16

Application Information

Patent Timeline

02 Jul 2019

Priority

16 Aug 2019

Application

CN201980060989.6

28 Oct 2025

Publication

CN112739375B

IPC: A61K39/00; C07K16/28; C07K16/00; C07K7/08; C40B40/10; C40B40/08

CPC: C07K16/2833; C07K16/30; C07K2317/34; C07K2317/32; C07K2317/565; C40B40/08; C40B40/10; C12N15/1037

Inventors: K·朱斯; G·J·A·雷尼

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Peptide libraries Tumor rejection antigen precursors

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AI Technical Summary

Technical Problem

Existing technologies have difficulty effectively targeting and binding complexes of specific HLA molecules and peptides, which limits the specific attack of immunotherapy on tumor cells.

Method used

An isolated antigen-binding protein (ABP) was developed that can bind to specific HLA subtypes (such as B*35:01 and A*01:01) and specifically bind to HLA-restricted peptides of HLA class I molecules, including specific amino acid positions and helical regions, enhancing targeting.

Benefits of technology

It improves the targeting and specific recognition capabilities of immunotherapy agents for tumor cells, and enhances the immune system's attack on tumor cells.

✦ Generated by Eureka AI based on patent content.

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Abstract

This article provides HLA-peptide targets and antigen-binding proteins that bind to HLA-peptide targets. Methods for identifying HLA-peptide targets and for identifying one or more antigen-binding proteins that bind to a given HLA-peptide target are also disclosed.

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Description

[0001] Cross-references

[0002] This application claims the benefits of U.S. Provisional Application No. 62 / 719,565, filed August 17, 2018; U.S. Provisional Application No. 62 / 808,775, filed February 21, 2019; and U.S. Provisional Application No. 62 / 869,923, filed July 2, 2019, which are incorporated herein by reference in their entirety.

[0003] Related applications

[0004] This application relates to PCT / US2018 / 046997, filed August 17, 2018, and PCT / US2018 / 06793, filed December 28, 2018, which are incorporated herein by reference in their entirety.

[0005] sequence list

[0006] This application includes a sequence list submitted electronically in ASCII format, which is hereby incorporated in its entirety by reference. The ASCII copy was created on September 30, 2019, named GSO-021WO_SL.txt, and has a size of 362,045 bytes. Background Technology

[0007] The immune system employs two types of adaptive immune responses to provide antigen-specific protection against pathogens: humoral immune responses and cellular immune responses, which specifically recognize pathogen antigens through B lymphocytes and T lymphocytes, respectively.

[0008] As antigen-specific effectors of cell-mediated immunity, T lymphocytes play a central role in the body's defense against diseases mediated by intracellular pathogens (such as viruses, intracellular bacteria, mycoplasma, and intracellular parasites) and in combating cancer cells through direct cytolysis of affected cells. The specificity of the T lymphocyte response is conferred and activated by the binding of the T cell receptor (TCR) to MHC molecules (the major histocompatibility complex) on the surface of affected cells. The T cell receptor is an antigen-specific receptor cloned on individual T lymphocytes, whose antigen-specific repertoire is generated via somatic gene rearrangement mechanisms, similar to those involved in the production of antibody gene repertoires. The T cell receptor comprises a heterodimer of transmembrane molecules, primarily consisting of α-β polypeptide dimers and a smaller subset of γ-δ polypeptide dimers. The T lymphocyte receptor subunit contains variable and constant regions similar to immunoglobulins within its extracellular domain, a short hinge region with cysteine residues promoting α and β chain pairing, a transmembrane region, and a short cytoplasmic region. TCR-triggered signal transduction is indirectly mediated by CD3-ζ, which is a related multi-subunit complex containing signal transduction subunits.

[0009] T lymphocyte receptors typically do not recognize natural antigens, but rather complexes displayed on the cell surface, including intracellularly processed antigen fragments that associate with the major histocompatibility complex (MHC) used to present peptide antigens. The MHC gene exhibits high polymorphism across species populations, comprising multiple common alleles for each individual gene. In humans, the MHC is known as human leukocyte antigen (HLA).

[0010] Major histocompatibility complex (MHC) class I molecules are expressed on the surface of almost all nucleated cells in vivo and are dimer molecules containing a transmembrane heavy chain (including a peptide antigen-binding groove) and a smaller extracellular chain called β2-microglobulin. The peptides presented by MHC class I molecules are derived from the degradation of cytoplasmic proteins by the proteasome, a multi-subunit structure in the cytoplasm (Niedermann G., 2002. Curr Top Microbiol Immunol. 268:91-136; for the processing of bacterial antigens, see Wick MJ and Ljunggren HG., 1999. Immunol Rev. 172:153-62). The cleaved peptide is transported to the endoplasmic reticulum (ER) lumen via antigen-processing-associated transporters (TAPs), where it binds to the groove of the assembled class I molecule. The resulting MHC / peptide complex is then transported to the cell membrane, enabling 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 peptide can be loaded onto MHC class I molecules in a TAP-independent manner and can also present extracellular proteins via cross-presentation. Thus, once the structure (peptide sequence and MHC isotype) of the complex is determined, a given MHC / peptide complex presents a novel protein structure on the cell surface that can be targeted by novel antigen-binding proteins (e.g., antibodies or TCRs).

[0011] Tumor cells can express antigens and display these antigens on their surface. These tumor-associated antigens can be used to develop novel immunotherapeutic agents that specifically target tumor cells. For example, tumor-associated antigens can be used to identify therapeutic antigen-binding proteins, such as TCRs, antibodies, or antigen-binding fragments. These tumor-associated antigens can also be used in pharmaceutical compositions, such as vaccines. Summary of the Invention

[0012] This article provides an isolated antigen-binding protein (ABP) that specifically binds to a human leukocyte antigen (HLA) peptide target, wherein the HLA peptide target comprises 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 molecules are HLA subtype B*35:01 (reference sequence: MGSHSMRYFYTAMSRPGRGEPRFIAVGYVDDTQFVRFDSDAASPRTEPRAPWIEQEGPEYWDRNTQIFKTNTQTYRESLRNLRGYYNQSEAGSHIIQRMYGCDLGPDGRLLRGHDQSAYDGKDYIALNEDLSSWTAADTAAQITQRKWEAARVAEQLRAYLEGLCVEWLRRYLENGKETLQRADPPKTHVTHHPVSDHEATLRCWALGFYPAEITLTWQRDGEDQTQDTELVETRPAGDRTFQKWAAVVVPSGEEQRYTCHVQHEGLPKPLTLR (SEQ ID NO:1)), and the HLA-restricted peptide comprises the sequence EVDPIGHVY (SEQ ID NO:2), and wherein the ABP is bound to any one or more of the following: (a) the restriction peptide EVDPIGHVY (SEQ ID NO:2) (b) any one or more of amino acid positions 2-9 of the α1 helix of HLA subtype B*35:01; and (c) any one or more of amino acid positions 147 and 148 of the α2 helix of HLA subtype B*35:01. Note that the enumerated range includes terminal residues. For example, an ABP symptom of any one or more of positions 2-9 of the restriction peptide EVDPIGHVY (SEQ ID NO:2) contacts at least one of residues 2, 3, 4, 5, 6, 7, 8, and 9 of the restriction peptide EVDPIGHVY (SEQ ID NO:2).

[0013] In some embodiments, the ABP binds to any one or more of amino acid positions 2-8 of the restriction peptide EVDPIGHVY (SEQ ID NO:2).

[0014] In some embodiments, the ABP binds to any one or more of amino acid positions 5-9 of the restriction peptide EVDPIGHVY (SEQ ID NO:2).

[0015] In some embodiments, the HLA class I molecule is HLA subtype B*35:01, and the HLA-restricted peptide consists of the sequence EVDPIGHVY (SEQ ID NO:2).

[0016] In some embodiments, the ABP comprises CDR-H3, which comprises a sequence selected from the following: CARDGVRYYGMDVW (SEQ ID NO:3), CARDGVRGYDRSAGYW (SEQ ID NO:4), CARDHDYGDYGEYFQHW (SEQ ID NO:5), CARDSWYCSSTSCGVNWFDPW (SEQ ID NO:6), CAKVNWNDGPYFDYW (SEQ ID NO:7), CATPTNSGYYGPYYYYGMDVW (SEQ ID NO:8), CARDVMDVW (SEQ ID NO:9), CAREGYGMDVW (SEQ ID NO:10), CARDNGVGVDYW (SEQ ID NO:11), CARGIADSGSYYGNGRDYYYGMDVW (SEQ ID NO:12), CARDDYYFDYW (SEQ ID NO:13), CARDGTRYYGMDVW (SEQ ID NO:14), CARDGVRYYGMDVW (SEQ ID NO:15), CARDGVRYYGMDVW (SEQ ID NO:16), CARDGVRYYGMDVW (SEQ ID NO:17), CARDGVRYYGMDVW (SEQ ID NO:18), CARDGVRYYGMDVW (SEQ ID NO:19), CARDGVRYYGMDVW (SEQ ID NO:10), CARDGVRYYGMDVW (SEQ ID NO:11), CARDGVRYYGMDVW (SEQ ID NO:12), CARDGVRYYGMDVW (SEQ ID NO:13), CARDGVRYYGMDVW (SEQ ID NO:14), CARDGVRYYGMDVW (SEQ ID NO:15), CARDGVRYYGMDVW (SEQ ID NO:16), CARDGVRYYGMDVW (SEQ ID NO:17), CARDGVRYYGMDVW (SEQ ID NO:18), CARDGVRYYGMDVW (SEQ ID NO:1 NO:14), CARDVVANFDYW (SEQ ID NO:15), CARGHSSGWYYYYGMDVW (SEQ ID NO:16), CAKDLGSYGGYYW (SEQ ID NO:17), CARSWFGGFNYHYYGMDVW (SEQ ID NO:18), CARELPIGYGMDVW (SEQ ID NO:19) and CARGGSYYYYGMDVW (SEQ ID NO:20).

[0017] In some embodiments, the ABP includes CDR-L3, which comprises sequences selected from the following: CMQGLQTPITF (SEQ ID NO:21), CMQALQTPPTF (SEQ ID NO:22), CQQAISFPLTF (SEQ ID NO:23), CQQANSFPLTF (SEQ ID NO:24), CQQANSFPLTF (SEQ ID NO:24), CQQSYSIPLTF (SEQ ID NO:25), CQQTYMMPYTF (SEQ ID NO:26), CQQSYITPWTF (SEQ ID NO:27), CQQSYITPYTF (SEQ ID NO:28), CQQYYTTPYTF (SEQ ID NO:29), CQQSYSTPLTF (SEQ ID NO:30), CMQALQTPLTF (SEQ ID NO:31), CQQYGSWPRTF (SEQ ID NO:32), CQQSYSTPVTF (SEQ ID NO:33), CMQALQTPYTF (SEQ ID NO:34), CQQSYSTPLTF ...35), CQQSYSTPLTF (SEQ ID NO:36), CQQSYSTPLTF (SEQ ID NO:37), CQQSYSTPVTF (SEQ ID NO:38), CQQSYSTPLTF (SEQ ID NO:39), CQQSYSTPLTF (SEQ ID NO:30), CQQALQTPLTF (SEQ ID NO:31), CQQYGSWPRTF (SEQ ID NO:32), CQQSYSTPVTF ( ID NO:34), CQQANSFPFTF (SEQ ID NO:35), CMQALQTPLTF (SEQ ID NO:31) and CQQSYSTPLTF (SEQ ID NO:30).

[0018] In some implementations, the ABP includes CDR-H3 and CDR-L3 from scFv, which are named G5_P7_E7, G5_P7_B3, G5_P7_A5, G5_P7_F6, G5-P1B12, G5-P1C12, G5-P1-E05, G5-P3G01, G5-P3G08, G5-P4B02, G5-P4E04, G5R4-P1D06, G5R4-P1H11, G5R4-P2B10, G5R4-P2H8, G5R4-P3G05, G5R4-P4A07, or G5R4-P4B01.

[0019] In some implementations, the ABP comprises all three heavy chain CDRs and all three light chain CDRs from the scFv, which are named G5_P7_E7, G5_P7_B3, G5_P7_A5, G5_P7_F6, G5-P1B12, G5-P1C12, G5-P1-E05, G5-P3G01, G5-P3G08, G5-P4B02, G5-P4E04, G5R4-P1D06, G5R4-P1H11, G5R4-P2B10, G5R4-P2H8, G5R4-P3G05, G5R4-P4A07, or G5R4-P4B01.

[0020] In some embodiments, the ABP comprises a VH sequence selected from: QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYDINWVRQAPGQGLEWMGIINPRSGSTKYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGVRYYGMDVWGQGTTVTVSS (SEQ ID NO:36), QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSHDINWVRQAPGQGLEWMGWMNPNSGDTGYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGVRGYDRSAGYWGQGTLVIVSS (SEQ ID NO:37), EVQLLESGGGLVKPGGSLRLSCAASGFSFSSYWMSWVRQAPGKGLEWISYISGDSGYTNYADSVKGRFTISRDDSKNTLYLQMNSLKTEDTAVYYCASHDYGDYGEYFQHWGQGTLVTVSS (SEQ ID NO:38), EVQLLQSGGGLVQPGGSLRLSCAASGFTFSNSDMNWVRQAPGKGLEWVAYISSGSSTIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVSWYCSSTSCGVNWFDPWGQGTLVTVSS (SEQ ID NO:39), EVQLLESGGGLVQPGGSLRLSCAASGFTFSNSDMNWVRQAPGKGLEWVASISSSGGYINYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKVNWNDGPYFDYWGQGTLVTVSS (SEQ ID NO:40), QVQLVQSGAEVKKPGSSVKVSCKASGGTFSNFGVSWLRQAPGQGLEWMGGIIPILGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCATPTNSGYYGPYYYYGMDVWGQGTTVTVSS (SEQ ID NO:41), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDVMDVWGQGTTVTVSS (SEQ IDNO:42)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSGYLVSWVRQAPGQGLEWMGWINPNSGGTNTAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYCAREGYGMDVWGQGTTVTVSS(SEQ ID NO:43)、QVQLVQSGAEVKKPGASVKVSCKASGYIFRNYPMHWVRQAPGQGLEWMGWINPDSGGTKYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYCARDNGVGVDYWGQGTLVTVSS(SEQ ID NO:44)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWMNPNIGNTGYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGIADSGSYYGNGRDYYYGMDVWGQGTTVTVSS(SEQ ID NO:45), QVQLVQSGAEVKKPGASVKVSCKASGGTFSSYGISWVRQAPGQGLEWMGWINPNSGVTKYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGDYYFDYWGQGTLVTVSS(SEQ ID NO:46), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYDINWVRQAPGQGLEWMGWINPNSGDTKYSQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGTRYYGMDVWGQGTTVTVSS(SEQ ID NO:47), EVQLLESGGGLVKPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVSYISSSSSYTNYADSVKGRFTISRDDSKNTLYLQMNSLKTEDTAVYYCARDVVANFDYWGQGTLVTVSS(SEQ ID NO:48) NO:48)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGWMNPDSGSTGYAQRFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGHSSGWYYYYGMDVWGQGTTVTVSS(SEQ IDNO: 49), EVQLLESGGGLVQPGGSLRLSCAASGFTFTSYSMHWVRQAPGKGLEWVSSITSFTNTMYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDLGSYGGYYWGQGTLVTVSS (SEQ ID NO: 50), QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARSWFGGFNYHYYGMDVWGQGTTVTVSS (SEQ ID NO: 51), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGWMNPNSGNTGYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARELPIGYGMDVWGQGTTVTVSS (SEQ ID NO: 52) and QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIVGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGGSYYYYGMDVWGQGTTVTVSS (SEQ ID NO: 53).

[0021] In some embodiments, the ABP comprises a VL selected from: DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSYRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGLQTPITFGQGTRLEIK (SEQ ID NO:54), DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSSRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTPPTFGPGTKVDIK (SEQ ID NO:55), DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQAISFPLTFGQSTKVEIK (SEQ ID NO:56), DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYSASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPLTFGGGTKVEIK (SEQID NO:57), DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPLTFGGGTKVEIK (SEQ ID NO:58), DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSIPLTFGGGTKVEIK (SEQ ID NO:59), DIQMTQSPSSLSASVGDRVTITCRASQGISNYLNWYQQKPGKAPKLLIYYASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTYMMPYTFGQGTKVEIK (SEQ ID NO:60), DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYGASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYITPWTFGQGTKVEIK (SEQID NO:61)、DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYITPYTFGQGTKLEIK(SEQID NO:62)、DIVMTQSPDSLAVSLGERATINCKTSQSVLYRPNNENYLAWYQQKPGQPPKLLIYQASIREPGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYTTPYTFGQGTKLEIK(SEQ ID NO:63)、DIQMTQSPSSLSASVGDRVTITCRASQSISRFLNWYQQKPGKAPKLLIYGASRPQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGQGTKVEIK(SEQ ID NO:64)、DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSHRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTPLTFGGGTKVEIK(SEQ IDNO:65), EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYAASARASGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYGSWPRTFGQGTKVEIK(SEQ ID NO:66), DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYGASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQQANSFPFTFGPGTKVDIK(SEQ ID NO:67), DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTPYTFGQGTKVEIK(SEQ ID NO:68), DIQMTQSPSSLSASVGDRVTITCQASEDISNHLNWYQQKPGKAPKLLIYDALSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPFTFGPGTKVDIK(SEQ ID NO:69)NO:69), DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTPLTFGQGTKVEIK(SEQ ID NO:70) and DIQMTQSPSSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIK (SEQ ID NO:71).

[0022] In some implementations, the ABP comprises VH and VL sequences from scFv, which are named G5_P7_E7, G5_P7_B3, G5_P7_A5, G5_P7_F6, G5-P1B12, G5-P1C12, G5-P1-E05, G5-P3G01, G5-P3G08, G5-P4B02, G5-P4E04, G5R4-P1D06, G5R4-P1H11, G5R4-P2B10, G5R4-P2H8, G5R4-P3G05, G5R4-P4A07, and G5R4-P4B01.

[0023] This article also provides isolated antigen-binding proteins (ABPs) that specifically bind to human leukocyte antigen (HLA) peptide targets, wherein the HLA peptide targets include HLA-restricted peptides complexed with HLA class I molecules, wherein the HLA-restricted peptides are located in the peptide-binding groove of the α1 / α2 heterodimeric portion of the HLA class I molecule. Class I molecules are HLA subtype A*01:01 (reference sequence: MGSHSMRYFFTSVSRPGRGEPRFIAVGYVDDTQFVRFDSDAASQKMEPRAPWIEQEGPEYWDQETRNMKAHSQTDRANLGTLRGYYNQSEDGSHTIQIMYGCDVGPDGRFLRGYRQDAYDGKDYIALNEDLRSWTAADMAAQITKRKWEAVHAAEQRRVYLEGRCVDGLRRYLENGKETLQRTDPPKTHMTHHPISDHEATLRCWALGFYPAEITLTWQRDGEDQTQDTELVETRPAGDGTFQKWAAVVVPSGEEQRYTCHVQHEGLPKPLTLR (SEQ ID NO:72)), and the HLA-restricted peptide comprises the sequence NTDNNLAVY (SEQ ID NO:73), and wherein the ABP is bound to any one or more of the following: (a) the restriction peptide NTDNNLAVY (SEQ ID NO:73) (b) any one or more of residues 3-9 of the α1 helix of the HLA subtype allele A*01:01, and (c) any one or more of residues 140-160 of the α2 helix of the HLA subtype allele A*01:01.

[0024] In some embodiments, the ABP binds to any one or more residues 6-9 of the restricted peptide NTDNNLAVY (SEQ ID NO:73).

[0025] In some embodiments, the ABP binds to one or more residues 7-8 of the restricted peptide NTDNNLAVY (SEQ ID NO:73).

[0026] In some embodiments, the ABP binds to any one or more residues 157-160 of the α2 helix of the HLA subtype allele A*01:01.

[0027] In some embodiments, the ABP binds to any one or more of residues 6-9 of the restriction peptide NTDNLAVY (SEQ ID NO:73) and any one or more of residues 157-160 of the α2 helix of the HLA subtype allele A*01:01.

[0028] In some embodiments, the HLA class I molecule is HLA subtype A*01:01, and the HLA-restricted peptide consists of the sequence NTDNNLAVY (SEQ ID NO:73).

[0029] In some embodiments, the ABP comprises a CDR-H3, and the CDR-H3 comprises a sequence selected from the following: CAATEWLGVW (SEQ ID NO:74), CARANWLDYW (SEQ ID NO:75), CARANWLDYW (SEQ ID NO:75), CARDWVLDYW (SEQ ID NO:76), CARGEWLDYW (SEQ ID NO:77), CARGWELGYW (SEQ ID NO:78), CARDFVGYDDW (SEQ ID NO:79), CARDYGDLDYW (SEQ ID NO:80), CARGSYGMDVW (SEQ ID NO:81), CARDGYSGLDVW (SEQ ID NO:82), CARDSGVGMDVW (SEQ ID NO:83), CARDGVAVASDYW (SEQ ID NO:84), CARGVNVDDFDYW (SEQ ID NO:85), CARGDYTGNWYFDLW (SEQ ID NO:86), CARANWLDYW (SEQ ID NO:75), CARDQFYGGNSGGHDYW (SEQ ID NO:87), CAREEDYW (SEQ ID NO:88), CARGDWFDPW (SEQ ID NO:89), CARGDWFDPW (SEQ ID NO:89), CARGEWFDPW (SEQ ID NO:90), CARSDWFDPW (SEQ ID NO:91), CARDSGSYFDYW (SEQ ID NO:92), CARDYGGYVDYW (SEQ ID NO:93), CAREGPAALDVW (SEQ ID NO:94), CARERRSGMDVW (SEQ ID NO:95), CARVLQEGMDVW (SEQ ID NO:96), CASERELPFDIW (SEQ ID NO:97), CAKGGGGYGMDVW (SEQ ID NO:98), CAAMGIAVAGGMDVW (SEQ ID NO:99), CARNWNLDYW (SEQ ID NO:100), CATYDDGMDVW (SEQ ID NO:101), CARGGGGALDYW (SEQ ID NO:102), CALSGNYYGMDVW (SEQ ID NO:103), CARGNPWELRLDYW (SEQ ID NO:104) and CARDKNYYGMDVW (SEQ ID NO:105).

[0030] In some embodiments, the ABP comprises a CDR-L3, and the CDR-L3 comprises a sequence selected from the following: CQQSYNTPYTF (SEQ ID NO:106), CQQSYSTPYTF (SEQ ID NO:107), CQQSYSTPYSF (SEQ ID NO:108), CQQSYSTPFTF (SEQ ID NO:109), CQQSYGVPYTF (SEQ ID NO:110), CQQSYSAPYTF (SEQ ID NO:111), CQQSYSAPYTF (SEQ ID NO:111), CQQSYSAPYSF (SEQ ID NO:112), CQQSYSTPYTF (SEQ ID NO:107), CQQSYSVPYSF (SEQ ID NO:113), CQQSYSAPYTF (SEQ ID NO:111), CQQSYSVPYSF (SEQ ID NO:113), CQQSYSTPQTF (SEQ ID NO:114), CQQLDSYPFTF (SEQ ID NO:115), CQQSYSSPYTF (SEQ ID NO:116), CQQSYSTPLTF (SEQ ID NO:30), CQQSYSTPYSF (SEQ ID NO:108), CQQSYSTPYTF (SEQ ID NO:107), CQQSYSTPYTF (SEQ ID NO:107), CQQSYSTPFTF (SEQ ID NO:109), CQQSYSTPTF (SEQ ID NO:117), CQQTYAIPLTF (SEQ ID NO:118), CQQSYSTPYTF (SEQ ID NO:107), CQQSYIAPFTF (SEQ ID NO:119), CQQSYSIPLTF (SEQ ID NO:25), CQQSYSNPTF (SEQ ID NO:120), CQQSYSTPYSF (SEQ ID NO:108), CQQSYSDQWTF (SEQ ID NO:121), CQQSYLPPYSF (SEQ ID NO:122), CQQSYSSPYTF (SEQ ID NO:116), CQQSYTTPWTF (SEQ ID NO:123), CQQSYLPPYSF (SEQ ID NO:122), CQEGITYTF (SEQ ID NO:124), CQQYYSYPFTF (SEQ ID NO:125), and CQHYGYSPVTF (SEQ ID NO:126).

[0031] In some implementations, the ABP includes CDR-H3 and CDR-L3 from scFv, which are named G2-P1H11, G2-P2E07, G2-P2E03, G2-P2A11, G2-P2C06, G2-P1G01, G2-P1C02, G2-P1H01, G2-P1B12, G2-P1B06, G2-P2H10, G2-P1H10, G2-P2C11, G2-P1C09, and G2-P1A10. G2-P1B10, G2-P1D07, G2-P1E05, G2-P1D03, G2-P1G12, G2-P2H11, G2-P1C03, G2-P1G07, G2-P1F12, G2-P1G03, G2-P2B08, G2-P2A10, G2-P2D04, G2-P1C06, G2-P2A09, G2-P1B08, G2-P1E03, G2-P2A03, G2-P2F01 or G2-P1D06.

[0032] In some implementations, the ABP includes CDR-H3 and CDR-L3 from scFv named G2-P1H11.

[0033] In some implementations, the ABP comprises all three heavy chain CDRs and all three light chain CDRs from the scFv, which are named G2-P1H11, G2-P2E07, G2-P2E03, G2-P2A11, G2-P2C06, G2-P1G01, G2-P1C02, G2-P1H01, G2-P1B12, G2-P1B06, G2-P2H10, G2-P1H10, G2-P2C11, G2-P1C09, G2-P1 A10, G2-P1B10, G2-P1D07, G2-P1E05, G2-P1D03, G2-P1G12, G2-P2H11, G2-P1C03, G2-P1G07, G2-P1F12, G2-P1G03, G2-P2B08, G2-P2A10, G2-P2D04, G2-P1C06, G2-P2A09, G2-P1B08, G2-P1E03, G2-P2A03, G2-P2F01 or G2-P1D06.

[0034] In some implementations, the ABP comprises all three heavy chain CDRs and all three light chain CDRs from the scFv named G2-P1H11.

[0035] In some embodiments, the ABP comprises a VH sequence selected from: QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYMHWVRQAPGQGLEWMGMINPSGGGTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGNPWELRLDYWGQGTLVTVSS (SEQ ID NO:127), QVQLVQSGAEVKKPGASVKVSCKASGGTFSSATISWVRQAPGQGLEWMGWIYPNSGGTVYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAATEWLGVWGQGTTVTVSS (SEQ ID NO:128), EVQLLQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGWINPNSGGTISAPNFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARANWLDYWGQGTLVTVSS (SEQ ID NO:129), EVQLLESGAEVKKPGASVKVSCKASGYTFTTYDLAWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARANWLDYWGQGTLVTVSS (SEQ ID NO:130), QVQLVQSGAEVKKPGASVKVSCKSSGYSFDSYVVNWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDWVLDYWGQGTLVTVSS (SEQ ID NO:131), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWMNPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGEWLDYWGQGTLVTVSS (SEQID NO:132), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGWELGYWGQGTLVTVSS (SEQ IDNO:133)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTINWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDFVGYDDWGQGTLVTVSS(SEQ ID NO:134)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGITWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDYGDLDYWGQGTLVTVSS(SEQ ID NO:135)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSNYILSWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSYGMDVWGQGTTVTVSS(SEQ ID NO:136)、QVQLVQSGAEVKKPGASVKVSCKASGYSFTRYNMHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGYSGLDVWGKGTTVTVSS(SEQ ID NO:137)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGWINPNNGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDSGVGMDVWGQGTTVTVSS(SEQ ID NO:138)、QVQLVQSGAEVKKPGASVKVSCKASGGTFNNYAFSWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGVAVASDYWGQGTLVTVSS(SEQ ID NO:139)、QVQLVQSGAEVKKPGASVKVSCKASGYTFSSYNMHWVRQAPGQGLEWMGWINGNTGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGVNVDDFDYWGQGTLVTVSS(SEQIDNO:140), QVQLVQSGAEVKKPGASVKVSCKASGGTFSSYAFSWVRQAPGQGLEWMGWINPDTGYTRYAQFQGRVTMTRDTSTVYMELSSLRSEDTAVYYCARGDYTGNWYFDLWGRGTLVTVSS(SEQ ID NO:141), EVQLLESGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWINPYSGGTNYAQKLQGRVTMTRDTSTVYMELSSLRSEDTAVYYCARANWLDYWGQGTLVTVSS(SEQ ID NO:142), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWISAYNGYTNYAQNLQGRVTMTRDTSTVYMELSSLRSEDTAVYYCARDQFYGGNSGGHDYWGQGTLVTVSS(SEQ ID NO:143) NO:143)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYNMHWVRQAPGQGLEWMGWMNPNSSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYCAREEDYWGQGTLVTVSS(SEQID NO:144)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTINWVRQAPGQGLEWMGWINPNSGGANYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGDWFDPWGQGTLVTVSS(SEQ ID NO:145)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQGLEWMGWISPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGDWFDPWGQGTLVTVSS(SEQ ID NO:146)、QVQLVQSGAEVKKPGASVKVSCKASGYTFSDYYVHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYCARGEWFDPWGQGTLVTVSS(SEQ IDNO:147)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTTYYMHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARSDWFDPWGQGTLVTVSS(SEQID NO:148)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSNYAINWVRQAPGQGLEWMGWISPYSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDSGSYFDYWGQGTLVTVSS(SEQ ID NO:149)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYYMHWVRQAPGQGLEWMGWIYPNTGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDYGGYVDYWGQGTLVTVSS(SEQ ID NO:150)、EVQLLESGAEVKKPGASVKVSCKASGYTFTSYAMNWVRQAPGQGLEWMGWMNPNSGGTKYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAREGPAALDVWGQGTLVTVSS(SEQ ID NO:151)、QVQLVQSGAEVKKPGASVKVSCKASGYTLTSHLIHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARERRSGMDVWGQGTTVTVSS(SEQ ID NO:152)、EVQLLESGAEVKKPGASVKVSCKASGYSFTDYIVHWVRQAPGQGLEWMGWINPYSGGTKYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARVLQEGMDVWGQGTLVTVSS(SEQ ID NO:153)、QVQLVQSGAEVKKPGASVKVSCKASGYTFSNFLINWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCASERELPFDIWGQGTMVTVSS(SEQ IDNO:154), QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYQMFWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKGGGGYGMDVWGQGTTVTVSS (SEQ ID NO:155), QVQLVQSGAEVKKPGASVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAAMGIAVAGGMDVWGQGTLVTVSS (SEQ ID NO:156), QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYHMHWVRQAPGQGLEWMGWIHPDSGGTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARNWNLDYWGQGTLVTVSS (SEQ ID NO:157), QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWMNPNSGNTGYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCATYDDGMDVWGQGTTVTVSS (SEQ ID NO:158), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYTVNWVRQAPGQGLEWMGWINPNSGGTKYAQNFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGGGGALDYWGQGTLVTVSS (SEQ ID NO:159), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGMINPRDDTTDYARDFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCALSGNYYGMDVWGQGTTVTVSS (SEQ ID NO:160) and QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSQYMHWVRQAPGQGLEWMGRIIPLLGIVNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDKNYYGMDVWGQGTTVTVSS (SEQ ID NO:161).

[0036] In some embodiments, the ABP comprises a VL sequence selected from: DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYSYPFTFGPGTKVDIK (SEQ ID NO:162), DIQMTQSPSSLSASVGDRVTITCRASQSISTWLAWYQQKPGKAPKLLIYAASSLRSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYNTPYTFGQGTKLEIK (SEQ ID NO:163), DIQMTQSPSSLSASVGDRVTITCRASQSISRWLAWYQQKPGKAPKLLIYAASTVQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTKLEIK (SEQ ID NO:164), DIQMTQSPSSLSASVGDRVTITCRASQDISRWLAWYQQKPGKAPKLLIYAASRLQAGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYSFGQGTKLEIK (SEQ ID NO:165), DIQMTQSPSSLSASVGDRVTITCRASQTISSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKVDIK (SEQ ID NO:166), DIQMTQSPSSLSASVGDRVTITCRASQTISSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYGVPYTFGQGTKVEIK (SEQ ID NO:167), DIQMTQSPSSLSASVGDRVTITCRASQSISNWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSAPYTFGPGTKVDIK (SEQIDNO:168), DIQMTQSPSSLSASVGDRVTITCRASQSVGNWLAWYQQKPGKAPKLLIYGASSLQTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSAPYTFGQGTKVEIK(SEQ ID NO:169), DIQMTQSPSSLSASVGDRVTITCRASQNIGNWLAWYQQKPGKAPKLLIYAASTLQTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSAPYSFGQGTKLEIK(SEQ ID NO:170), DIQMTQSPSSLSASVGDRVTITCRASQSISRWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTKLEIK(SEQ ID NO:171), DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYGASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSVPYSFGQGTKLEIK(SEQ ID NO:172) NO:172)、DIQMTQSPSSLSASVGDRVTITCRASQSISKWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSAPYTFGQGTKVEIK(SEQ ID NO:173)、DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSVPYSFGQGTKLEIK(SEQ ID NO:173) ID NO:174)、DIQMTQSPSSLSASVGDRVTITCRASQTISNYLNWYQQKPGKAPKLLIYAASNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPQTFGQGTKVEIK(SEQ ID NO:175)、DIQMTQSPSSLSASVGDRVTITCRASRDIGRAVGWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQLDSYPFTFGPGTKVDIK(SEQ IDNO:176)、DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSSPYTFGPGTKVDIK(SEQID NO:177)、DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKLEIK(SEQ ID NO:178)、DIQMTQSPSSLSASVGDRVTITCRASQSIGRWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYSFGQGTKVEIK(SEQ ID NO:179)、DIQMTQSPSSLSASVGDRVTITCRASQSISTWLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFAQGTKLEIK(SEQ ID NO:180)、DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYGASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTKLEIK(SEQID NO:181)、DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKVDIK(SEQ ID NO:182)、DIQMTQSPSSLSASVGDRVTITCRASQSVSNWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPTFGQGTKLEIK(SEQ ID NO:183)、DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTYAIPLTFGGGTKVEIK(SEQ IDNO:184)、DIQMTQSPSSLSASVGDRVTITCQASQDIGSWLAWYQQKPGKAPKLLIYATSSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTKLEIK(SEQ ID NO:185)、DIQMTQSPSSLSASVGDRVTITCRASQGISRWLAWYQQKPGKAPKLLIYAASTLQPGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYIAPFTFGPGTKVDIK(SEQ ID NO:186)、DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASRLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSIPLTFGGGTKVEIK(SEQ ID NO:187)、DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYGVSSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSNPTFGQGTKVEIK(SEQ ID NO:188)、DIQMTQSPSSLSASVGDRVTITCRASQSISSWVAWYQQKPGKAPKLLIYGASNLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYSFGQGTKLEIK(SEQ ID NO:189)、DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSDQWTFGQGTKVEIK(SEQ ID NO:190)、DIQMTQSPSSLSASVGDRVTITCRASQSISRWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYLPPYSFGQGTKVEIK(SEQ ID NO:191)、DIQMTQSPSSLSASVGDRVTITCRASQSISNWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTYFTLTISSLQPEDFATYYCQQSYSSPYTFGQGTKLEIK(SEQ IDNO:192),DIQMTQSPSSSLSASVGDRVTITCRASQSISHYLNWYQQKPGKAPKLLIYGASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYTTPWTFGQGTRLEIK(SEQ ID NO:193),DIQMTQSPSSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYLPPYSFGQGTKLEIK(SEQID NO:194),DIQMTQSPSSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYGASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQEGITYTFGQGTKVEIK(SEQ ID NO:195) and EIVMTQSPATLSVSPGERATLSCRASQSVSRNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQHYGYSPVTFGQGTKLEIK (SEQ ID NO:196).

[0037] In some implementations, the ABP comprises VH and VL sequences from scFv, wherein the scFv is named G2-P1H11, G2-P2E07, G2-P2E03, G2-P2A11, G2-P2C06, G2-P1G01, G2-P1C02, G2-P1H01, G2-P1B12, G2-P1B06, G2-P2H10, G2-P1H10, G2-P2C11, G2-P1C09, G2-P1A10, G... 2-P1B10, G2-P1D07, G2-P1E05, G2-P1D03, G2-P1G12, G2-P2H11, G2-P1C03, G2-P1G07, G2-P1F12, G2-P1G03, G2-P2B08, G2-P2A10, G2-P2D04, G2-P1C06, G2-P2A09, G2-P1B08, G2-P1E03, G2-P2A03, G2-P2F01 or G2-P1D06.

[0038] In some implementations, the ABP comprises a VH sequence and a VL sequence from an scFv named G2-P1H11.

[0039] This article also provides isolated antigen-binding proteins (ABPs) that specifically bind to human leukocyte antigen (HLA) peptide targets, wherein the HLA peptide targets include HLA-restricted peptides complexed with HLA class I molecules, wherein the HLA-restricted peptides are located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, wherein the HLA... Class I molecules are HLA subtype A*02:01 (reference sequence: MGSHSMRYFFTSVSRPGRGEPRFIAVGYVDDTQFVRFDSDAASQRMEPRAPWIEQEGPEYWDGETRKVKAHSQTHRVDLGTLRGYYNQSEAGSHTVQRMYGCDVGSDWRFLRGYHQYAYDGKDYIALKEDLRSWTAADMAAQTTKHKWEAAHVAEQLRAYLEGTCVEWLRRYLENGKETLQRTDAPKTHMTHHAVSDHEATLRCWALSFYPAEITLTWQRDGEDQTQDTELVETRPAGDGTFQKWAAVVVPSGQEQRYTCHVQHEGLPKPLTLR (SEQ ID NO:197)), wherein the HLA-restricted peptide comprises the sequence AIFPGAVPAA (SEQ ID NO:198), and wherein the ABP is bound to any one or more of the following: (a) the restriction peptide AIFPGAVPAA (SEQ ID NO:198) (a) any one or more of amino acid positions 1-6 of the α1 helix of HLA subtype A*02:01, (b) any one or more of amino acid positions 46, 49, 55, 61, 74, 76, 77, 78, 81, and 84 of the α1 helix of HLA subtype A*02:01, (c) any one or more of amino acid positions 45-60, 66, 67, and 73 of the α1 helix of HLA subtype A*02:01, (d) the α2 helix of HLA subtype A*02:01. The amino acid positions of the helix are any one or more of 138, 145, 147, 152-156, 164, and 167, and the amino acid positions of (e)HLA subtype A*02:01 are any one or more of 56, 59, 60, 63, 64, 66, 67, 70, 73, 74, 132, 150-153, 155, 156, 158-160, 162-164, 166-168, 170, and 171.

[0040] In some embodiments, the ABP binds to any one or more of amino acid positions 1-5 of the restriction peptide AIFPGAVPAA (SEQ ID NO:198).

[0041] In some embodiments, the ABP is bound to one or both of amino acid positions 4 and 5 of the restriction peptide AIFPGAVPAA (SEQ ID NO:198).

[0042] In some embodiments, the ABP binds to one or both of amino acid positions 5 and 6 of the restriction peptide AIFPGAVPAA (SEQ ID NO:198).

[0043] In some embodiments, the ABP binds to amino acid position 6 of the restriction peptide AIFPGAVPAA (SEQ ID NO:198).

[0044] In some embodiments, the ABP binds to one or more of the amino acid positions 46, 49, 55, 66, 67, and 73 of the α1 helix of HLA subtype A*02:01.

[0045] In some embodiments, the ABP comprises a VH region containing complementary sites, the complementary sites comprising at least one, two, three, or four residues of Tyr32, Gly99, Asp100, and Tyr100A from the VH region as shown in the Kabat numbering sequence QVQLVQSGAEVKKPGASVKVSCKASGGTLSSYPINWVRQAPGQGLEWMGWISTYSGHADYAQKLQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARSYDYGDYLNFDYWGQGTLVTVSS (SEQ ID NO: 199).

[0046] In some embodiments, the ABP comprises a VH region containing complementary sites, the complementary sites comprising residues of the VH region as shown in the Kabat numbering sequence QVQLVQSGAEVKKPGASVKVSCKASGGTLSSYPINWVRQAPGQGLEWMGWISTYSGHADYAQKLQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARSYDYGDYLNFDYWGQGTLVTVSS (SEQ ID NO: 199): Thr28, Leu 29, Ser 30, Ser 31, Tyr 32, Pro 33, Trp 47, Trp50, Ser 52, Tyr53, Ser 54, His 56, Asp 58, Tyr 59, Gln 61, Gln 64, Asp 97, Tyr At least one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty, twelve, or twenty-two residues from 98, Gly99, Asp100, Tyr100A, Leu100B, and Asn100C.

[0047] In some embodiments, the complementary site comprises at least one, two, three, four, five, six, or seven residues selected from the VH region residues Ser30, Ser31, Tyr32, Tyr98, Gly99, Asp100, and Tyr100A, as numbered according to the Kabat numbering system.

[0048] In some embodiments, the ABP comprises a VL region containing complementary sites, the complementary sites comprising at least one, two, or three residues of Tyr32, Ser 91, and Tyr 92 from the VL region as shown in the Kabat numbering sequence DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSIPPTFGGGTKVDIK (SEQ ID NO:200).

[0049] In some embodiments, the ABP comprises a VL region containing complementary sites, the complementary sites comprising at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 residues of the VL region as shown in the Kabat numbering sequence DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSIPPTFGGGTKVDIK (SEQ ID NO:200) as selected from Asp1, Ser30, Asn31, Tyr32, Tyr49, Ala50, Ser53, Ser67, Ser91, Tyr92, Ser93, Ile94, and Pro95.

[0050] In some embodiments, the complementary site comprises at least one, two, three, four, five, or six residues from the VL region selected as Asp1, Asn31, Tyr32, Ser91, Tyr92, and Ile94, as numbered according to the Kabat numbering system.

[0051] In some implementations, the HLA class I molecule is HLA subtype A*02:01, and the HLA-restricted peptide consists of the sequence AIFPGAVPAA (SEQ ID NO:198).

[0052] In some embodiments, the ABP comprises CDR-H3, which comprises a sequence selected from the following: CARDDYGDYVAYFQHW (SEQ ID NO:201), CARDLSYYYGMDVW (SEQ ID NO:202), CARVYDFWSVLSGFDIW (SEQ ID NO:203), CARVEQGYDIYYYYYMDVW (SEQ ID NO:204), CARSYDYGDYLNFDYW (SEQ ID NO:205), CARASGSGYYYYYGMDVW (SEQ ID NO:206), CAASTWIQPFDYW (SEQ ID NO:207), CASNGNYYGSGSYYNYW (SEQ ID NO:208), CARAVYYDFWSGPFDYW (SEQ ID NO:209), CAKGGIYYGSGSYPSW (SEQ ID NO:210), CARGLYYMDVW (SEQ ID NO:201). NO:211), CARGLYGDYFLYYGMDVW (SEQ ID NO:212), CARGLLGFGEFLTYGMDVW (SEQ ID NO:213), CARDRDSSWTYYYYGMDVW (SEQ ID NO:214), CARGLYGDYFLYYGMDVW (SEQ ID NO:212), CARGDYYDSSGYYFPVYFDYW (SEQ ID NO:215) and CAKDPPFWSGHYYYYGMDVW (SEQ ID NO:216).

[0053] In some embodiments, the ABP includes CDR-L3, which comprises sequences selected from the following: CQQNYNSVTF (SEQ ID NO:217), CQQSYNTPWTF (SEQ ID NO:218), CGQSYSTPPTF (SEQ ID NO:219), CQQSYSAPYTF (SEQ ID NO:111), CQQSYSIPPTF (SEQ ID NO:220), CQQSYSAPYTF (SEQ ID NO:111), CQQHNSYPPTF (SEQ ID NO:221), CQQYSTYPITI (SEQ ID NO:222), CQQANSFPWTF (SEQ ID NO:223), CQQSHSTPQTF (SEQ ID NO:224), CQQSYSTPLTF (SEQ ID NO:30), CQQSYSTPLTF (SEQ ID NO:30), CQQTYSTPWTF (SEQ ID NO:225), CQQYGSSPYTF (SEQ ID NO:225), CQQYGSSPYTF (SEQ ID NO:217), CQQSYNTPWTF (SEQ ID NO:218), CQQSYSTPWTF (SEQ ID NO:219), CQQSYSAPYTF (SEQ ID NO:219), CQQSYSTPWTF (SEQ ID NO:222), CQQSYNTPWTF (SEQ ID NO:223), CQQSYSTPWTF (SEQ ID NO:224), CQQSYSTPLTF (SEQ ID NO:30), CQQTYSTPWTF (SEQ ID NO:225), CQQYGSSPYTF (SEQ ID NO:225), CQQYGSSPYTF (SEQ ID NO:225), CQQSYNTPW ... NO:226), CQQSHSTPLTF (SEQ ID NO:227), CQQANGFPLTF (SEQ ID NO:228) and CQQSYSTPLTF (SEQ ID NO:30).

[0054] In some implementations, the ABP includes CDR-H3 and CDR-L3 from scFv, which are named G8-P1A03, G8-P1A04, G8-P1A06, G8-P1B03, G8-P1C11, G8-P1D02, G8-P1H08, G8-P2B05, G8-P2E06, R3G8-P2C10, R3G8-P2E04, R3G8-P4F05, R3G8-P5C03, R3G8-P5F02, R3G8-P5G08, G8-P1C01, or G8-P2C11.

[0055] In some implementations, the ABP comprises all three heavy chain CDRs and all three light chain CDRs from the scFv, which are named G8-P1A03, G8-P1A04, G8-P1A06, G8-P1B03, G8-P1C11, G8-P1D02, G8-P1H08, G8-P2B05, G8-P2E06, R3G8-P2C10, R3G8-P2E04, R3G8-P4F05, R3G8-P5C03, R3G8-P5F02, R3G8-P5G08, G8-P1C01, or G8-P2C11.

[0056] In some embodiments, the ABP comprises a VH sequence selected from: QVQLVQSGAEVKKPGASVKVSCKASGGTFSRSAITWVRQAPGQGLEWMGWINPNSGATNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDDYGDYVAYFQHWGQGTLVTVSS (SEQ ID NO:229), QVQLVQSGAEVKKPGASVKVSCKASGYPFIGQYLHWVRQAPGQGLEWMGIINPSGDSATYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDLSYYYGMDVWGQGTTVTVSS (SEQ ID NO:230), QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYMHWVRQAPGQGLEWMGWMNPIGGGTGYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARVYDFWSVLSGFDIWGQGTLVTVSS(SEQID NO:231), EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVSGINWNGGSTGYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVEQGYDIYYYYYMDVWGKGTTVTVSS (SEQ ID NO:232), QVQLVQSGAEVKKPGASVKVSCKASGGTLSSYPINWVRQAPGQGLEWMGWISTYSGHADYAQKLQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARSYDYGDYLNFDYWGQGTLVTVSS (SEQ ID NO:199), EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYWMSWVRQAPGKGLEWVSSISGRGDNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARASGSGYYYYYGMDVWGQGTTVTVSS (SEQ ID NO:233), QVQLVQSGAEVKKPGASVKVSCKASGYTFGNYFMHWVRQAPGQGLEWMGMVNPSGGSETFAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAASTWIQPFDYWGQGTLVTVSS (SEQ IDNO:234)、EVQLLESGGGLVQPGGSLRLSCAASGFDFSIYSMNWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASNGNYYGSGSYYNYWGQGTLVTVSS(SEQ ID NO:235)、QVQLVQSGAEVKKPGASVKVSCKASGYTLTTYYMHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARAVYYDFWSGPFDYWGQGTLVTVSS(SEQ ID NO:236)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGWINPYSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKGGIYYGSGSYPSWGQGTLVTVSS(SEQ ID NO:237)、QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYGVSWVRQAPGQGLEWMGWISPYSGNTDYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGLYYMDVWGKGTTVTVSS(SEQ ID NO:238)、QVQLVQSGAEVKKPGASVKVSCKASGYTFSNMYLHWVRQAPGQGLEWMGWINPNTGDTNYAQTFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGLYGDYFLYYGMDVWGQGTKVTVSS(SEQ ID NO:239)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGWMNPNSGNTGYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGLLGFGEFLTYGMDVWGQGTLVTVSS(SEQ ID NO:240)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYIHWVRQAPGQGLEWMGVINPSGGSTTYAQKLQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDRDSSWTYYYYGMDVWGQGTTVTVSS(SEQID NO: 241), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSNYMHWVRQAPGQGLEWMGWMNPNSGNTGYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGLYGDYFLYYGMDVWGQGTTVTVSS (SEQ ID NO: 242), QVQLVQSGAEVKKPGASVKVSCKASGGTFSSHAISWVRQAPGQGLEWMGVIIPSGGTSYTQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGDYYDSSGYYFPVYFDYWGQGTLVTVSS (SEQ ID NO: 243) and QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYAMNWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDPFWSGHYYYYGMDVWGQGTTVTVSS (SEQ ID NO: 244).

[0057] In some embodiments, the ABP comprises a VL sequence selected from: DIQMTQSPSSLSASVGDRVTITCRASQSITSYLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQNYNSVTFGQGTKLEIK (SEQ ID NO:245), DIQMTQSPSSLSASVGDRVTITCWASQGISSYLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYNTPWTFGPGTKVDIK (SEQ ID NO:246), DIQMTQSPSSLSASVGDRVTITCRASQAISNSLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCGQSYSTPPTFGQGTKLEIK (SEQ ID NO:247), DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYKASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSAPYTFGPGTKVDIK (SEQ ID NO:248), DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSIPPTFGGGTKVDIK (SEQ ID NO:200), DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSAPYTFGGGTKVEIK (SEQ ID NO:249), DIQMTQSPSSLSASVGDRVTITCRASQGINSYLAWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHNSYPPTFGQGTKLEIK (SEQ IDNO:250)、DIQMTQSPSSLSASVGDRVTITCRASQSISRWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTYPITIGQGTKVEIK(SEQ ID NO:251)、DIQMTQSPSSLSASVGDRVTITCRASQGISNSLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPWTFGQGTKLEIK(SEQ ID NO:252)、DIQMTQSPSSLSASVGDRVTITCRASQDVSTWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSHSTPQTFGQGTKVEIK(SEQ ID NO:253)、DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKLEIK(SEQ ID NO:254)、DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIK(SEQID NO:255)、DIQMTQSPSSLSASVGDRVTITCRASQGISNWLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTYSTPWTFGQGTKLEIK(SEQ ID NO:256)、EIVMTQSPATLSVSPGERATLSCRASQSVGNSLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYGSSPYTFGQGTKVEIK(SEQ ID NO:257)、DIQMTQSPSSLSASVGDRVTITCRASQSISGYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSHSTPLTFGQGTKVEIK(SEQ IDNO:258),DIQMTQSPSSSLSASVGDRVTITCRASQNIYTYLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANGFPLTFGGGTKVEIK(SEQ ID NO:259) and DIQMTQSPSSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIK (SEQ ID NO:71).

[0058] In some implementations, the ABP comprises a VH sequence and a VL sequence from an scFv, which is named G8-P1A03, G8-P1A04, G8-P1A06, G8-P1B03, G8-P1C11, G8-P1D02, G8-P1H08, G8-P2B05, G8-P2E06, R3G8-P2C10, R3G8-P2E04, R3G8-P4F05, R3G8-P5C03, R3G8-P5F02, R3G8-P5G08, G8-P1C01, or G8-P2C11.

[0059] This article also provides an isolated antigen-binding protein (ABP) that specifically binds to human leukocyte antigen (HLA)-peptide targets, wherein the HLA-peptide targets comprise HLA-restricted peptides complexed with HLA class I molecules, wherein the HLA-restricted peptides are located in the HLA... In the peptide-binding groove of the α1 / α2 heterodimer portion of class I molecules, wherein the HLA class I molecule is HLA subtype A*01:01 (reference sequence: MGSHSMRYFFTSVSRPGRGEPRFIAVGYVDDTQFVRFDSDAASQKMEPRAPWIEQEGPEYWDQETRNMKAHSQTDRANLGTLRGYYNQSEDGSHTIQIMYGCDVGPDGRFLRGYRQDAYDGKDYIALNEDLRSWTAADMAAQITKRKWEAVHAAEQRRVYLEGRCVDGLRRYLENGKETLQRTDPPKTHMTHHPISDHEATLRCWALGFYPAEITLTWQRDGEDQTQDTELVETRPAGDGTFQKWAAVVVPSGEEQRYTCHVQHEGLPKPLTLR (SEQ ID NO:72)), and the HLA-restricted peptide comprises the sequence ASSLPTTMNY (SEQ ID NO:72). NO:260), and wherein the ABP is bound to any one or more of the following: (a) any one or more of amino acid positions 4, 6, 7, 8 and 9 of the restriction peptide ASSLPTTMNY (SEQ ID NO:260), (b) any one or more of amino acid positions 49-56 of HLA subtype A*01:01, (c) any one or more of amino acid positions 59-66 of HLA subtype A*01:01, (d) any one or more of amino acid positions 136-147 of HLA subtype A*01:01, and (e) any one or more of amino acid positions 157-160 of HLA subtype A*01:01.

[0060] In some embodiments, the ABP binds to any one or more of amino acid positions 6-9 of the restriction peptide ASSLPTTMNY (SEQ ID NO:260).

[0061] In some embodiments, the ABP binds to any one or more of amino acid positions 6-7 in the restriction peptide ASSLPTTMNY (SEQ ID NO:260).

[0062] In some embodiments, the ABP binds to amino acid position 6 of the restricted peptide ASSLPTTMNY (SEQ ID NO:260).

[0063] In some embodiments, the ABP is combined with: (a) any one or more of amino acid positions 52-54 of HLA subtype A*01:01, (b) any one or more of amino acid positions 136-139 of HLA subtype A*01:01, (c) any one or more of amino acid positions 141-147 of HLA subtype A*01:01, or (d) any one or more of amino acid positions 136-139 and any one or more of amino acid positions 141-147 of HLA subtype A*01:01.

[0064] In some embodiments of an ABP containing an antibody or an antigen-binding fragment thereof, the HLA class I molecule is HLA subtype A*01:01, and the HLA-restricted peptide consists of the sequence ASSLPTTMNY (SEQ ID NO:260).

[0065] In some embodiments, the ABP comprises CDR-H3, which comprises a sequence selected from the following: CARDQDTIFGVVITWFDPW (SEQ ID NO:261), CARDKVYGDGFDPW (SEQ ID NO:262), CARDEDDSMDVW (SEQ ID NO:263), CARDSSGLDPW (SEQ ID NO:264), CARGVGNLDYW (SEQ ID NO:265), CARDHAHQYYDFWSGYYSGTYYYGMDVW (SEQ ID NO:266), CAREQWPSYWYFDLW (SEQ ID NO:267), CADRGYSYGYFDYW (SEQ ID NO:268), CARGSGDPNYYYYYGLDVW (SEQ ID NO:269), CARDTGDHFDYW (SEQ ID NO:270), CARAENGMDVW (SEQ ID NO:271), CARDPGGYMDVW (SEQ ID NO:269), CARDTGDHFDYW (SEQ ID NO:270), CARAENGMDVW (SEQ ID NO:271), CARDPGGYMDVW (SEQ ID NO:262), CARDGGYMDVW (SEQ ID NO:263), CARDGGYMDVW (SEQ ID NO:264), CARDGGYMDVW (SEQ ID NO:265), CARDGHQYYDFWSGYYSGTYYYGMDVW (SEQ ID NO:266), CARDGHQYYDFWSGYYFDLW (SEQ ID NO:267), CARDGHQYYFDLW (SEQ ID NO:268), CARDGHQYYYFDLW (SEQ ID NO:269), CARDGHQYYFDLW (SEQ ID NO:260), CARDGHQYYFDLW (SEQ ID NO:269), CARDGHQYYFDLW (SEQ ID NO:260), CARDGHQYYFDLW (SEQ ID NO: ID NO:272), CARDGDAFDIW (SEQ ID NO:273), CARDMGDAFDIW (SEQ ID NO:274), CAREEDGMDVW (SEQ ID NO:275), CARDTGDHFDYW (SEQ ID NO:270), CARGEYSSGFFFVGWFDLW (SEQ ID NO:276) and CARETGDDAFDIW (SEQ ID NO:272) NO:277).

[0066] In some embodiments, the ABP includes CDR-L3, which comprises sequences selected from the following: CQQYFTTPYTF (SEQ ID NO:278), CQQAEAFPYTF (SEQ ID NO:279), CQQSYSTPITF (SEQ ID NO:280), CQQSYIIPYTF (SEQ ID NO:281), CHQTYSTPLTF (SEQ ID NO:282), CQQAYSFPWTF (SEQ ID NO:283), CQQGYSTPLTF (SEQ ID NO:284), CQQANSFPRTF (SEQ ID NO:285), CQQANSLPYTF (SEQ ID NO:286), CQQSYSTPFTF (SEQ ID NO:109), CQQSYSTPFTF (SEQ ID NO:109), CQQSYGVPTF (SEQ ID NO:287), CQQSYSTPLTF (SEQ ID NO:30), CQQSYSTPLTF (SEQ ID NO:287). NO:30), CQQYYSYPWTF (SEQ ID NO:288), CQQSYSTPFTF (SEQ ID NO:109), CMQTLKTPLSF (SEQ ID NO:289) and CQQSYSTPLTF (SEQ ID NO:30).

[0067] In some implementations, the ABP includes CDR-H3 and CDR-L3 from scFv, which are named R3G10-P1A07, R3G10-P1B07, R3G10-P1E12, R3G10-P1F06, R3G10-P1H01, R3G10-P1H08, R3G10-P2C04, R3G10-P2G11, R3G10-P3E04, R3G10-P4A02, R3G10-P4C05, R3G10-P4D04, R3G10-P4D10, R3G10-P4E07, R3G10-P4E12, R3G10-P4G06, R3G10-P5A08, or R3G10-P5C08.

[0068] In some implementations, the ABP comprises all three heavy chain CDRs and all three light chain CDRs from the scFv, which are named R3G10-P1A07, R3G10-P1B07, R3G10-P1E12, R3G10-P1F06, R3G10-P1H01, R3G10-P1H08, and R3G10-P2C. 04, R3G10-P2G11, R3G10-P3E04, R3G10-P4A02, R3G10-P4C05, R3G10-P4D04, R3G10-P4D10, R3G10-P4E07, R3G10-P4E12, R3G10-P4G06, R3G10-P5A08 or R3G10-P5C08.

[0069] In some embodiments, the ABP comprises a VH sequence selected from: EVQLLESGGGLVKPGGSLRLSCAASGFTFSSYWMSWVRQAPGKGLEWVSGISARSGRTYYADSVKGRFTISRDDSKNTLYLQMNSLKTEDTAVYYCARDQDTIFGVVITWFDPWGQGTLVTVSS (SEQ ID NO:290), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIIHPGGGTTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDKVYGDGFDPWGQGTLVTVSS (SEQ ID NO:291), QVQLVQSGAEVKKPGASVKVSCKASGYIFTGYYMHWVRQAPGQGLEWMGMIGPSDGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAREDDSMDVWGKGTTVTVSS(SEQ IDNO:292), QVQLVQSGAEVKKPGASVKVSCKASGYTFIGYYMHWVRQAPGQGLEWMGMIGPSDGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDSSGLDPWGQGTLVTVSS (SEQ ID NO:293), QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGMIGPSDGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGVGNLDYWGQGTLVTVSS (SEQ ID NO:294), QVQLVQSGAEVKKPGASVKVSCKASGVTFSTSAISWVRQAPGQGLEWMGWISPYNGNTDYAQMLQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDAHQYYDFWSGYYSGTYYYGMDVWGQGTTVTVSS(SEQ ID NO:295), QVQLVQSGAEVKKPGASVKVSCKASGGTFSNSIINWVRQAPGQGLEWMGWMNPNSGNTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAREQWPSYWYFDLWGRGTLVTVSS(SEQ IDNO:296), QVQLVQSGAEVKKPGASVKVSCKASGGTFSTHDINWVRQAPGQGLEWMGVINPSGGSAIYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDRGYSYGYFDYWGQGTLVTVSS(SEQ ID NO:297), QVQLVQSGAEVKKPGASVKVSCKASGNTFIGYYVHWVRQAPGQGLEWVGIINPNGGSISYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSGDPNYYYYYGLDVWGQGTTVTVSS(SEQ ID NO:298), QVQLVQSGAEVKKPGASVKVSCKASGYTLSYYYMHWVRQAPGQGLEWMGMIGPSDGSTSYAQRFQGRVTMTRDTSTGTVYMELSSLRSEDTAVYYCARDTGDHFDYWGQGTLVTVSS(SEQ ID NO:299) NO:299)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGIIGPSDGSTTYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYCARAENGMDVWGQGTTVTVSS(SEQ ID NO:300)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYVHWVRQAPGQGLEWMGIIAPSDGSTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDPGGYMDVWGKGTTVTVSS(SEQ ID NO:301)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYLHWVRQAPGQGLEWMGMIGPSDGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGDAFDIWGQGTMVTVSS(SEQ ID NO:302)、QVQLVQSGAEVKKPGSSVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGRISPSDGSTTYAPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDMGDAFDIWGQGTTVTVSS(SEQ IDNO: 303), QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGMIGPSDGSTSYAQRFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAREEDGMDVWGQGTTVTVSS (SEQ ID NO: 304), QVQLVQSGAEVKKPGASVKVSCKASGYTLSYYYMHWVRQAPGQGLEWMGMIGPSDGSTSYAQRFQGRVTMTRDTSTGTVYMELSSLRSEDTAVYYCARDTGDHFDYWGQGTLVTVSS (SEQ ID NO: 299), QVQLVQSGAEVKKPGSSVKVSCKASGGTFNNFAISWVRQAPGQGLEWMGGIIPIFDATNYAQKFQGRVTFTADESTSTAYMELSSLRSEDTAVYYCARGEYSSGFFFVGWFDLWGRGTQVTVSS (SEQ ID NO: 305) and QVQLVQSGAEVKKPGASVKVSCKASGYNFTGYYMHWVRQAPGQGLEWMGIIAPSDGSTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARETGDDAFDIWGQGTMVTVSS (SEQ ID NO: 306).

[0070] In some embodiments, the ABP comprises a VL sequence selected from: DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYFTTPYTFGQGTKLEIK (SEQ ID NO:307), DIQMTQSPSSLSASVGDRVTITCRASQSISRWLAWYQQKPGKAPKLLIFDASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQAEAFPYTFGQGTKVEIK (SEQ ID NO:308), DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPITFGQGTRLEIK (SEQ ID NO:309), DIQMTQSPSSLSASVGDRVTITCRASQSISNYLNWYQQKPGKAPKLLIYKASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYIIPYTFGQGTKLEIK (SEQ ID NO:310), DIQMTQSPSSLSASVGDRVTITCRASQSISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQTYSTPLTFGQGTKVEIK (SEQ ID NO:311), DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYSASNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQAYSFPWTFGQGTKVEIK (SEQ ID NO:312), DIQMTQSPSSLSASVGDRVTITCRASQNISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYSTPLTFGQGTRLEIK (SEQIDNO:313), DIQMTQSPSSLSASVGDRVTITCRASQDISRYLAWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPRTFGQGTKVEIK(SEQ ID NO:314), DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSLPYTFGQGTKVEIK(SEQ ID NO:315), DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASTLQNGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKVDIK(SEQ ID NO:316), DIQMTQSPSSLSASVGDRVTITCRASQRISSYLNWYQQKPGKAPKLLIYSASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKVDIK(SEQ ID NO:317) NO:317)、DIQMTQSPSSLSASVGDRVTITCRASQSISSYLAWYQQKPGKAPKLLIYDASKLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYGVPTFGQGTKLEIK(SEQ ID NO:318)、DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIK(SEQ ID NO:319)、DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIK(SEQ ID NO:319) NO:71)、DIQMTQSPSSLSASVGDRVTITCRASQGISTYLAWYQQKPGKAPKLLIYDASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYSYPWTFGQGTRLEIK(SEQ IDNO:320),DIQMTQSPSSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASTLQNGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKVDIK(SEQ ID NO:316), DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQTLKTPLSFGGTKVEIK(SEQ ID NO:321) and DIQMTQSPSSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIK (SEQ ID NO:71).

[0071] In some implementations, the ABP comprises a VH sequence and a VL sequence from an scFv, which is named R3G10-P1A07, R3G10-P1B07, R3G10-P1E12, R3G10-P1F06, R3G10-P1H01, R3G10-P1H08, R3G10-P2C04, R3G10-P2G11, R3G10-P3E04, R3G10-P4A02, R3G10-P4C05, R3G10-P4D04, R3G10-P4D10, R3G10-P4E07, R3G10-P4E12, R3G10-P4G06, R3G10-P5A08, or R3G10-P5C08.

[0072] This article also provides an isolated antigen-binding protein (ABP) that specifically binds to human leukocyte antigen (HLA) peptide targets, wherein the HLA peptide targets include HLA-restricted peptides complexed with HLA class I molecules, wherein the HLA-restricted peptides are located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule, and wherein the HLA... Class I molecules are HLA subtype A*02:01 (reference sequence: MGSHSMRYFFTSVSRPGRGEPRFIAVGYVDDTQFVRFDSDAASQRMEPRAPWIEQEGPEYWDGETRKVKAHSQTHRVDLGTLRGYYNQSEAGSHTVQRMYGCDVGSDWRFLRGYHQYAYDGKDYIALKEDLRSWTAADMAAQTTKHKWEAAHVAEQLRAYLEGTCVEWLRRYLENGKETLQRTDAPKTHMTHHAVSDHEATLRCWALSFYPAEITLTWQRDGEDQTQDTELVETRPAGDGTFQKWAAVVVPSGQEQRYTCHVQHEGLPKPLTLR (SEQ ID NO: 197)), and the HLA-restricted peptide comprises the sequence LLASSILCA (SEQ ID NO: 322), and wherein the ABP is bound to any one or more of the following: (a) the restricted peptide LLASSILCA (SEQ ID NO: 322) (a) any one or more of residues 1-5 of the HLA-A*02:01α1 helix, (b) any one or more of residues 49-85 of the HLA-A*02:01α1 helix, and (c) any one or more of residues 57-67 of the HLA-A*02:01α1 helix.

[0073] In some embodiments of the ABP comprising an antibody or an antigen-binding fragment thereof, the HLA class I molecule is HLA subtype A*02:01, and the HLA-restricted peptide consists of the sequence LLASSILCA (SEQ ID NO:322).

[0074] In some embodiments, the ABP comprises CDR-H3, which comprises a sequence selected from the following: CARDGYDFWSGYTSDDYW (SEQ ID NO:323), CASDYGDYR (SEQ ID NO:324), CARDLMTTVVTPGDYGMDVW (SEQ ID NO:325), CARQDGGAFAFDIW (SEQ ID NO:326), CARELGYYYGMDVW (SEQ ID NO:327), CAALIFGVPLLPYGMDVW (SEQ ID NO:328), CAKDLATVGEPYYYYGMDVW (SEQ ID NO:329), and CARLWFGELHYYYYYGMDVW (SEQ ID NO:330).

[0075] In some embodiments, the ABP includes CDR-L3, which comprises sequences selected from the following: CHHYGRSHTF (SEQ ID NO:331), CQQANAFPPTF (SEQ ID NO:332), CQQYYSIPLTF (SEQ ID NO:333), CQQSYSTPPTF (SEQ ID NO:334), CQQSYSFPYTF (SEQ ID NO:335), CMQALQTPLTF (SEQ ID NO:31), CQQGNTFPLTF (SEQ ID NO:336), and CMQGSHWPPSF (SEQ ID NO:337).

[0076] In some implementations, the ABP includes CDR-H3 and CDR-L3 from scFv, which are named G7R3-P1C6, G7R3-P1G10, 1-G7R3-P1B4, 2-G7R4-P2C2, 3-G7R4-P1A3, 4-G7R4-B5-P2E9, 5-G7R4-B10-P1F8 or B7 (G7R3-P3A9).

[0077] In some implementations, the ABP comprises all three heavy chain CDRs and all three light chain CDRs from the scFv, which is named G7R3-P1C6, G7R3-P1G10, 1-G7R3-P1B4, 2-G7R4-P2C2, 3-G7R4-P1A3, 4-G7R4-B5-P2E9, 5-G7R4-B10-P1F8 or B7 (G7R3-P3A9).

[0078] In some embodiments, the ABP comprises a VH sequence selected from the following: QVQLVQSGAEVKKPGASVKVSCKASGGTFSNYGISWVRQAPGQGLEWMGIINPGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGYDFWSGYTSDDYWGQGTLVTVSS (SEQ ID NO:338), EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMHWVRQAPGKGLEWVSGISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASDYGDYRGQGTLVTVSS (SEQ ID NO:339), QVQLVQSGAEVKKPGASVKVSCKASGYTFSNYYIHWVRQAPGQGLEWMGWLNPNSGNTGYAQRFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDLMTTVVTPGDYGMDVWGQGTTVTVSS (SEQ ID NO:340), QVQLVQSGAEVKKPGASMKVSCKASGYTFTTDGISWVRQAPGQGLEWMGRIYPHSGYTEYAKKFKGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARQDGGAFAFDIWGQGTMVTVSS (SEQ ID NO:341), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSQYMHWVRQAPGQGLEWMGWISPNNGDTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARELGYYYGMDVWGQGTTVTVSS (SEQ ID NO:342), QVQLVQSGAEVKKPGSSVKVSCKASRYTFTSYDINWVRQAPGQGLEWMGRIIPMLNIANYAPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARALIFGVPLLPYGMDVWGQGTTVTVSS (SEQ ID NO:343), EVQLLQSGGGLVQPGGSLRLSCAASGFTFSSSWMHWVRQAPGKGLEWVSFISTSSGYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDLATVGEPYYYYGMDVWGQGTTVTVSS (SEQ IDNO: 344) and QVQLVQSGAEVKKPGSSVKVSCKASGDTFNTYALSWVRQAPGQGLEWMGWMNPNSGNAGYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARLWFGELHYYYYYGMDVWGQGTMVTVSS (SEQ ID NO: 345).

[0079] In some embodiments, the ABP comprises a VL sequence selected from: EIVMTQSPATLSVSPGERATLSCRASQSVSSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCHHYGRSHTFGQGTKVEIK (SEQ ID NO:346), DIQMTQSPSSLSASVGDRVTITCRASQDIRNDLGWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANAFPPTFGQGTKVEIK (SEQ ID NO:347), DIVMTQSPDSLAVSLGERATINCKSSQSVFYSSNNKNQLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSIPLTFGQGTKLEIK (SEQ ID NO:348), DIQMTQSPSSLSASVGDRVTITCQASQDIFKYLNWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPTFGQGTRLEIK (SEQ ID NO:349), DIQMTQSPSSLSASVGDRVTITCRASQSISTWLAWYQQKPGKAPKLLIYYASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSFPYTFGQGTKVEIK (SEQ ID NO:350), DIVMTQSPLSLPVTPGEPASISCSSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTPLTFGGGTKVEIK (SEQ ID NO:351), DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYSASNLRSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGNTFPLTFGQGTKVEIK (SEQIDNO:352) and DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGSHWPPSFGQGTRLEIK (SEQ ID NO:353).

[0080] In some implementations, the ABP comprises a VH sequence and a VL sequence from an scFv, which is named G7R3-P1C6, G7R3-P1G10, 1-G7R3-P1B4, 2-G7R4-P2C2, 3-G7R4-P1A3, 4-G7R4-B5-P2E9, 5-G7R4-B10-P1F8, or B7 (G7R3-P3A9).

[0081] In some embodiments, the ABP comprises an antibody or an antigen-binding fragment thereof. In some embodiments, the antigen-binding protein is linked to a scaffold, optionally the scaffold comprises serum albumin or Fc, optionally wherein the Fc is a human Fc and is an isotype of IgG (IgG1, IgG2, IgG3, IgG4), IgA (IgA1, IgA2), IgD, IgE, or IgM. 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 a hinge region of a human antibody. In some embodiments, the antigen-binding protein comprises an Fv fragment, a Fab fragment, an F(ab')2 fragment, a Fab' fragment, a scFv fragment, a scFv-Fc fragment, and / or a single-domain antibody or an antigen-binding fragment thereof. In some embodiments, the antigen-binding protein comprises an scFv fragment. In some embodiments, the antigen-binding protein comprises one or more antibody complementarity-determining regions (CDRs), optionally six antibody CDRs. In some embodiments, the antigen-binding protein comprises an antibody. In some embodiments, the antigen-binding protein is a monoclonal antibody. In some embodiments, the antigen-binding protein is a humanized, human, or chimeric antibody. In some embodiments, the antigen-binding protein is multispecific, optionally bispecific. In some embodiments, the antigen-binding protein binds to more than one antigen or more than one epitope on a single antigen. In some embodiments, the antigen-binding protein comprises a heavy chain constant region selected from classes of IgG, IgA, IgD, IgE, and IgM. In some embodiments, the antigen-binding protein comprises a heavy chain constant region of class human IgG and subclasses selected from IgG1, IgG4, IgG2, and IgG3. In some embodiments, the antigen-binding protein comprises one or more modifications that extend its half-life. In some embodiments, the antigen-binding protein comprises a modified Fc, optionally containing one or more mutations that extend its half-life, optionally YTE.

[0082] In some embodiments of the isolated ABP, the ABP comprises a T cell receptor (TCR) or its antigen-binding portion. In some embodiments, the TCR or its antigen-binding portion comprises a TCR variable region. In some embodiments, the TCR or its antigen-binding portion comprises one or more TCR complementarity-determining regions (CDRs).

[0083] In some embodiments, the TCR comprises α-chains and β-chains. In some embodiments, the TCR comprises γ-chains and δ-chains.

[0084] In some embodiments, the antigen-binding protein is part of a chimeric antigen receptor (CAR) comprising: an extracellular portion containing the antigen-binding protein; and an intracellular signaling domain. In some embodiments, the antigen-binding protein comprises scFv, and the intracellular signaling domain comprises an immune receptor tyrosine activation motif (ITAM). In some embodiments, the intracellular signaling domain comprises a signaling domain of the ζ chain of the CD3-ζ (CD3) chain.

[0085] In some embodiments, the ABP further includes a transmembrane domain connecting the extracellular domain and the intracellular signal transduction domain. In some embodiments, the transmembrane domain includes the transmembrane portion of CD28.

[0086] In some embodiments, the ABP further comprises an intracellular signaling domain of a T-cell co-stimulatory molecule. In some embodiments, the T-cell co-stimulatory molecule is CD28, 4-1BB, OX-40, ICOS, or any combination thereof.

[0087] This article also provides isolated polynucleotides encoding the isolated ABP as described herein.

[0088] In some embodiments of the ABP, the antigen-binding protein binds to the HLA-peptide target via contact sites with HLA class I molecules and with HLA-restricted peptides of the HLA-peptide target. In some embodiments of the ABP, the binding of the ABP to amino acid positions on the restriction peptide or HLA subtype, or the contact sites or residues that directly or indirectly affect the binding of the HLA-peptide target to the ABP, are determined by position scanning, hydrogen-deuterium exchange, or protein crystallography.

[0089] In some embodiments, the ABP can be used as a drug. In some embodiments, the ABP can be used to treat cancer, optionally wherein the cancer expresses or is expected to express an HLA-peptide target. In some embodiments, the ABP can be used to treat cancer, wherein the cancer is selected from solid tumors and hematologic malignancies.

[0090] This article also provides ABP, a conserved modified variant of the ABP described herein. This article also provides antigen-binding proteins (ABPs) that compete with the antigen-binding proteins described herein for binding. This article also provides antigen-binding proteins (ABPs) that bind to the same HLA-peptide epitopes as the antigen-binding proteins described herein.

[0091] This document also provides engineered cells expressing receptors comprising the antigen-binding protein described herein. In some embodiments, the engineered cells are T cells, optionally cytotoxic T cells (CTLs). In some embodiments of the engineered cells, the antigen-binding protein is expressed from a heterologous promoter.

[0092] This article also provides isolated polynucleotides or polynucleotide sequences encoding the antigen-binding proteins or their antigen-binding moieties described herein.

[0093] This article also provides isolated polynucleotides or polynucleotide sequences encoding the HLA / peptide targets described herein.

[0094] This article also provides vectors or vector sets containing the polynucleotides or polynucleotide groups described herein.

[0095] This document also provides a host cell comprising the polynucleotides or polynucleotide groups described herein or the vectors or vector groups described herein, optionally wherein the host cell is CHO or HEK293, or optionally wherein the host cell is a T cell.

[0096] This article also provides methods for generating antigen-binding proteins, including: expressing the antigen-binding protein in host cells as described herein and isolating the expressed antigen-binding protein.

[0097] This article also provides pharmaceutical compositions comprising the antigen-binding protein described herein and pharmaceutically acceptable excipients.

[0098] This document also provides a method for treating a subject's cancer, comprising: administering to the subject an effective amount of the antigen-binding protein described herein or a pharmaceutical composition described herein, optionally wherein the cancer is selected from solid tumors and hematologic malignancies. In some embodiments, the cancer expresses or is expected to express an HLA-peptide target.

[0099] This article also provides kits that include the antigen-binding protein or pharmaceutical composition described herein, along with instructions for use.

[0100] This article also provides compositions comprising at least one HLA-peptide target and adjuvant described herein.

[0101] This article also provides compositions comprising at least one HLA-peptide target described herein and a pharmaceutically acceptable excipient.

[0102] This document also provides compositions comprising amino acid sequences including at least one HLA-peptide target polypeptide disclosed in Table A, Table A1 or Table A2, optionally, the amino acid sequence consisting substantially of or composed of the polypeptide.

[0103] This document also provides viruses comprising the isolated polynucleotides or polynucleotide sequences described herein. In some embodiments, the virus is a filamentous bacteriophage.

[0104] This article also provides yeast cells containing the isolated polynucleotides or polynucleotide groups described herein. Attached Figure Description

[0105] The following description and accompanying drawings will help to better understand these and other features, aspects and advantages of the invention, wherein:

[0106] Figure 1 This shows the general structure of human leukocyte antigen (HLA) class I molecules. A personal work by user atropos235 published via en.wikipedia, CC BY 2.5, https: / / commons.wikimedia.org / w / index.php?curid=1805424

[0107] Figure 2 An exemplary construct element for cloning TCRs into an expression system for therapeutic development is depicted.

[0108] Figure 3 The design of the target and micro-collection negative control for the HLA-peptide target “G5” is shown.

[0109] Figure 4 The design of the target and micro-collection negative control for HLA-peptide targets “G8” and “G10” is shown.

[0110] Figure 5 A and Figure 5 B shows the HLA stability results of the G5 reverse screening "micro-assemblies" and the G5 target. Figure 5 A discloses SEQ ID NOS 2, 358 and 378-379, Figure 5 B discloses SEQ ID NOS 2, 358, 378-379 and 636, all of which are disclosed in the order of their appearance.

[0111] Figure 6 A- Figure 6 E shows the HLA stability results of reverse-screening peptides from the “complete” G5 set. Figure 6 A discloses SEQ IDNOS 368-369 and 359, Figure 6 B disclosed SEQ ID NOS 370-371 and 360, Figure 6 C discloses SEQ ID NOS 380-381 and 637, Figure 6D disclosed SEQ ID NOS 372-374, Figure 6 E disclosed SEQ ID NOS 375-377, and the above disclosures are made in the order of their appearance.

[0112] Figure 7 A and Figure 7 B shows the HLA stability results for the reverse screening peptide and the G8 target. Figure 7 A discloses SEQ ID NOS198 and 638-640, which are disclosed in the order of their appearance.

[0113] Figure 8 A and Figure 8 B shows the HLA stability results of the G10 reverse screening "micro-assembly" and the G10 target. Figure 8 A discloses SEQ ID NOS 260, 370-371, and 360. Figure 8 B discloses SEQ ID NOS 260, 370-371, 360 and 641, all of which are disclosed in the order of their appearance.

[0114] Figure 9 A- Figure 9 D shows the HLA stability results of the reverse screening peptides from the additional G8 and G10 “complete” set. Figure 9 A disclosed SEQ ID NOS 368-369 and 359, Figure 9 B disclosed SEQ ID NOS 383-385, Figure 9 C discloses SEQ IDNOS 372-374, Figure 9 D disclosed SEQ ID NOS 380-382, and the above disclosures are made in the order of their appearance.

[0115] Figure 10 A- Figure 10 C shows the ELISA results of the phage supernatant, indicating that G5-, G8-, and G10-binding phages were gradually enriched as the panning wheel progressed.

[0116] Figure 11 A flowchart describing the antibody selection process is shown, including the standards and intended applications for scFv, Fab, and IgG forms.

[0117] Figure 12 A, Figure 12 B and Figure 12C depicts the biolayer interferometry (BLI) results of Fab clone G5-P7A05 to HLA peptide target B*35:01-EVDPIGHVY (SEQ ID NO:2), Fab clones R3G8-P2C10 and G8-P1C11 to HLA peptide target A*02:01-AIFPGAVPAA (SEQ ID NO:198), and Fab clone R3G10-P1B07 to HLA peptide target A*01:01-ASSLPTTMNY (SEQ ID NO:260).

[0118] Figure 13 The general experimental design for a position scanning experiment is shown.

[0119] Figure 14 A shows the stability results for the G5 position variant-HLA. Figure 14 A disclosed SEQ ID NO:2.

[0120] Figure 14 B shows the binding affinity of Fab clone G5-P7A05 to the G5 position variant-HLA. Figure 14 B disclosed SEQ ID NO:2.

[0121] Figure 15 A shows the stability results for the G8 positional variants HLAs. Figure 15 A disclosed SEQ ID NO:198.

[0122] Figure 15 B shows the binding affinity of Fab clone G8-P2C10 to the G8 position variant HLA. Figure 15 B disclosed SEQ ID NO:198.

[0123] Figure 16 A shows the stability results for the G10 position variant-HLA. Figure 16 A disclosed SEQ ID NO:260.

[0124] Figure 16 B shows the binding affinity of Fab clone G10-P1B07 to the G10 position variant HLA. Figure 16 B disclosed SEQ ID NO:260.

[0125] Figure 17 A, Figure 17 B and Figure 17 C shows a representative example of antibodies that bind to G5-, G8-, or G10-presenting K562 cells, as detected by flow cytometry.

[0126] Figure 18 A- Figure 18C shows a histogram of K562 cells binding to the generated target-specific antibodies.

[0127] Figure 19 A- Figure 19 C shows a histogram of cell binding assays using tumor cell lines expressing HLA subtypes and target genes of selected HLA-peptide targets.

[0128] Figure 20A and Figure 20B The number of target-specific T cells from the tested donors is shown in (A) and the number of target-specific unique TCR clones is shown in (B). Figure 20A SEQ ID NOS 260 and 367 have been disclosed. Figure 20B SEQ IDs NOS260 and 367 have been disclosed, and the disclosures are made in the order of their appearance.

[0129] Figure 21 A shows an exemplary heatmap of scFv G8-P1H08, which uses a comprehensive perturbation view to visualize the entire HLA portion of the HLA-peptide target G8. Figure 21 A disclosed SEQ ID NO:642. Figure 21 B shows an example of HDX data from scFv G8-P1H08 plotted on the crystal structure PDB5bs0 (1jf1.pd, available at http: / / www.rcsb.org / structure / 1JF1).

[0130] Figure 22 A shows a heatmap of the HLAα1 helix of all ABPs tested against the HLA-peptide target G8 (HLA-A*02:01_AIFPGAVPAA(SEQ ID NO:198)). Figure 22 A disclosed SEQ ID NO:643. Figure 22 B shows a heatmap of the HLAα2 helix of all ABPs tested against HLA-peptide target G8 (HLA-A*02:01_AIFPGAVPAA(SEQ ID NO:198)). Figure 22 B disclosed SEQ ID NO:644. Figure 22 C shows the resulting heatmap of the restricted peptide AIFPGAVPAA (SEQ ID NO: 198) of ABP for all tests. Figure 22 C disclosed SEQ ID NO:198.

[0131] Figure 23 A shows an exemplary heatmap of scFv R3G10-P2G11, which uses a comprehensive perturbation view to visualize the entire HLA portion of the HLA-peptide target G10. Figure 23 A disclosed SEQ ID NO:645.

[0132] Figure 23 B shows an example of HDX data from scFv R3G10-P2G11 plotted on the crystal structure PDB5bs0.

[0133] Figure 23 C shows an example of HDX data from scFv G10-P5A08 plotted on the crystal structure PDB5bs0. Figure 23 C discloses "RRVY" as SEQ ID NO:646.

[0134] Figure 24 A shows the resulting heatmap of the HLAα1 helix of all ABPs tested against the HLA-peptide target G10 (HLA-A*01:01_ASSLPTTMNY(SEQ ID NO:260)). Figure 24 A disclosed SEQ ID NO:647. Figure 24 B shows the resulting heatmap of the HLAα2 helix of all ABPs tested against HLA-peptide target G10 (HLA-A*01:01_ASSLPTTMNY(SEQ ID NO:260)). Figure 24 B disclosed SEQ ID NO:648. Figure 24 C shows the resulting heatmap of the restricted peptide ASSLPTTMNY (SEQ ID NO:260) of ABP for all tests.

[0135] Figure 25 Exemplary spectral data for the peptide EVDPIGHVY (SEQ ID NO:2) are depicted. This plot includes peptide fragmentation information and patient-sample-related information, including HLA type.

[0136] Figure 26 Exemplary spectral data for peptide AIFPGAVPAA (SEQ ID NO:198) is depicted. This plot includes peptide fragmentation information and patient-sample-related information, including HLA type.

[0137] Figure 27 Exemplary spectral data for the peptide ASSLPTTMNY (SEQ ID NO:260) are depicted. This plot includes peptide fragmentation information and patient-sample-related information, including HLA type.

[0138] Figure 28 A and 28B depict size exclusion chromatographic fractions (A) and SDS-PAGE analysis of chromatographic fractions under reducing conditions (B).

[0139] Figure 29 Micrographs depicting exemplary crystals of a complex comprising Fab clone G8-P1C11 and HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0140] Figure 30 The overall structure of the complex formed by binding G8-P1C11 to the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”) via Fab cloning was depicted.

[0141] Figure 31 Fine electron density regions of the crystal structure of Fab clone G8-P1C11, which is complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”), are depicted. The depicted regions correspond to the restriction peptide AIFPGAVPAA (SEQ ID NO:198).

[0142] Figure 32 LigPlot depicts the interaction between HLA and the restriction peptide. The crystal structure corresponds to the Fab clone G8-P1C11 complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0143] Figure 33 A diagram depicting the interacting residues between the Fab VH and VL chains and the restriction peptide is provided. The crystal structure corresponds to the Fab clone G8-P1C11 complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0144] Figure 34 LigPlot depicts the interaction between the restriction peptide chain and the Fab chain. The crystal structure corresponds to the Fab clone G8-P1C11 complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0145] Figure 35 LigPlot depicts the interaction between the Fab VH chain and HLA. The crystal structure corresponds to the Fab clone G8-P1C11 complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0146] Figure 36LigPlot depicts the interaction between the Fab VL chain and HLA. The crystal structure corresponds to the Fab clone G8-P1C11 complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0147] Figure 37 An interface overview of the PISA analysis depicting the interaction between HLA and the restriction peptide is provided. The crystal structure corresponds to the Fab clone G8-P1C11 complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0148] Figure 38 PISA analysis depicted the interaction residues between HLA and the restriction peptide. The crystal structure corresponds to Fab clone G8-P1C11 complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0149] Figure 39 The Fab VH chain and the PISA analysis of the interacting residues with the restriction peptide are described. The crystal structure corresponds to the Fab clone G8-P1C11 complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0150] Figure 40 PISA analysis depicted the interacting residues between the Fab VL chain and the restriction peptide. The crystal structure corresponds to the Fab clone G8-P1C11 complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0151] Figure 41 An interface overview of PISA analysis depicting the interaction between the Fab VH chain and HLA is provided. The crystal structure corresponds to the Fab clone G8-P1C11 complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0152] Figure 42 PISA analysis depicted the interaction residues between the Fab VH chain and HLA. The crystal structure corresponds to the Fab clone G8-P1C11 complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0153] Figure 43An interface overview of PISA analysis depicting the interaction between the Fab VL chain and HLA is provided. The crystal structure corresponds to the Fab clone G8-P1C11 complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0154] Figure 44 PISA analysis depicted the interaction residues between the Fab VL chain and HLA. The crystal structure corresponds to the Fab clone G8-P1C11 complexed with the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0155] Figure 45 A depicts an exemplary heatmap of the HLA portion of the G8 HLA-peptide complex when incubated with the scFv clone G8-P1C11, which visualizes the whole using a comprehensive perturbation. Figure 45 A disclosed SEQ ID NO:642.

[0156] Figure 45 B depicts an instance of HDX data of scFv G8-P1C11 plotted on the crystal structure of Fab clone G8-P1C11 complexed with HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0157] Figure 46 The binding affinity of Fab clone G8-P1C11 to the G8 position variant HLA was depicted. Figure 46 SEQ ID NO:198 has been disclosed.

[0158] Figure 47 Histograms of K562 cells bound to G8-P1C11 (a target-specific antibody against the HLA-peptide target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”)) are shown.

[0159] Figure 48 An exemplary construct backbone sequence for cloning TCRs into an expression system for therapeutic development is depicted. Figure 48 SEQ ID NO:650 has been disclosed.

[0160] Figure 49 An exemplary construct sequence is described for cloning a TCR specific to A*0201_LLASSILCA (SEQ ID NO:322) into an expression system for therapeutic development. Figure 49 SEQ ID NO:651 was also disclosed.

[0161] Figure 50 An exemplary construct sequence is described for cloning a TCR specific to A*0101_EVDPIGHLY (SEQ ID NO:354) into an expression system for therapeutic development. Figure 50 SEQ ID NO:652 was also disclosed.

[0162] Figure 51 The spectral data for peptide EVDPIGHLY (SEQ ID NO:354) are displayed. This figure includes peptide fragmentation information and patient-sample-related information, including HLA type.

[0163] Figure 52 The spectral data for peptide GVHGGILNK (SEQ ID NO:355) are displayed. This figure includes peptide fragmentation information and patient-sample-related information, including HLA type.

[0164] Figure 53 The spectral data of peptide GVYDGEEHSV (SEQ ID NO:356) are shown.

[0165] Figure 54 The spectral data of peptide NTDNLAVY (SEQ ID NO:73) are shown.

[0166] Figures 55-63 The spectral data for other peptides disclosed in Table A are shown. Figure 55 SEQ ID NO: 653, 653, and 653 have been disclosed. Figure 56 SEQ ID NOS 654, 654 and 654 have been disclosed. Figure 57 SEQ ID NOS 655, 655 and 655 have been disclosed. Figure 58 SEQ ID NOS 656, 656 and 656 have been disclosed. Figure 59 SEQ ID NOS 656, 656 and 656 have been disclosed. Figure 60 SEQ ID NOS 657, 657 and 657 have been disclosed. Figure 61 SEQ ID NOS 658, 658 and 658 have been disclosed. Figure 62 SEQ IDNOS 659, 659 and 659 are disclosed. Figure 63 SEQ ID NO 355, 355 and 355 are disclosed, and the disclosures are made in the order of their appearance.

[0167] Figure 64 The design of target screening 1 for the G2 target HLA-A*01:01_NTDNNLAVY (SEQ ID NO:73) is shown.

[0168] Figure 65 A shows the target and micro-collection negative control design for the G2 target. Figure 65 A discloses SEQ ID NOS 73, 370-371 and 360 in the order of their appearance.

[0169] Figure 65 B shows the stability ELISA results for the G2 reverse screening "mini-set" and the G2 target. Figure 65 B discloses SEQ ID NOS 73, 370-371, 360 and 641 in the order of their appearance.

[0170] Figure 66 The stability ELISA results for the additional G2 “complete” set of reverse-screening peptides are shown. Figure 66 SEQ ID NOS 368-369, 359, 372-377, 358 and 378-382 are disclosed in the order of their appearance.

[0171] Figure 67 The design of target screening 2 for the G7 target HLA-A*02:01_LLASSILCA (SEQ ID NO:322) is shown.

[0172] Figure 68 The stability ELISA results for the additional G7 “complete set” reverse screening peptides are shown. Figure 68 SEQ ID NOS 372-374, 380-382, 638-640, 383-385, 370-371 and 360 are disclosed in the order of their appearance.

[0173] Figure 69 A shows the target and micro-collection negative control design for the G7 target. Figure 69 A discloses SEQ ID NOS 322, 368-369 and 359 in the order of their appearance.

[0174] Figure 69 B shows the stability ELISA results for the G7 reverse screening "mini-set" and the G7 target. Figure 69 B discloses SEQ ID NOS 322, 368-369, 359 and 375 in the order of their appearance.

[0175] Figure 70 A and Figure 70 B shows the phage panning results for targets G2 and G7, respectively.

[0176] Figure 71 A and Figure 71B shows the biolayer interferometry (BLI) results for the G2 target Fab clone G-2P1H11 and the G7 target G7R4-B5-P2E9, respectively.

[0177] Figure 72 The amino acid substitution map of the position scanning experiment described in this paper is shown. Figure 72 SEQ ID NOS 73 and 322 are disclosed in the order of their appearance.

[0178] Figure 73 A shows a stability heatmap of the G2 position variant - HLA. Figure 73 A disclosed SEQ ID NO:73.

[0179] Figure 73 B shows an affinity heatmap of Fab clone G2-P1H11. Figure 73 B disclosed SEQ ID NO:73.

[0180] Figure 74 A shows a stability heatmap of the G7 position variant. Figure 74 A disclosed SEQ ID NO:322.

[0181] Figure 74 B shows an affinity heatmap of Fab clone G7R4-B5-P2E9. Figure 74 B disclosed SEQ ID NO:322.

[0182] Figure 75 The results show the cell binding of Fab clones G2-P1H11 and G7R4-B5-P2E9 to HLA-transduced K562 cells bombarded with the target or negative control peptides.

[0183] Figure 76 The results show the cell binding of Fab clones G2-P1H11 and G7R4-B5-P2E9 to HLA-transduced K562 cells bombarded with the target or negative control peptides.

[0184] Figure 77 An example of hydrogen-deuterium exchange (HDX) data plotted on the crystal structure PDB 5bs0 is shown.

[0185] Figure 78 An exemplary HDX heatmap of scFv clone G2-P1G07 is shown, which is visualized as a whole using a comprehensive perturbation view. Figure 78 SEQ ID NO:660 has been disclosed.

[0186] Figure 79 HDX heatmaps of the tested G2 scFv and Fab clones across the HLA α1 and α2 helices are shown. Figure 79 SEQ ID NOS 661-662 are disclosed in the order of their appearance.

[0187] Figure 80 HDX heatmaps of the tested G2 scFv and Fab clone restricted peptide NTDNNLAVY (SEQ ID NO:73) are shown.

[0188] Figure 81 An experimental workflow is described, which allows for the isolation of TCRs that specifically bind to HLA-peptide targets.

[0189] Figure 82 The flow cytometry sorting procedure for sorting MHC target-specific CD8+ T cells is shown. Figure 82 "GEMSSNSTAL" is disclosed as SEQ ID NO:357 and "EVDPIGHLY" is disclosed as SEQ ID NO:354.

[0190] Figure 83 The flow cytometry results for exemplary HLA-peptide targets B*44:02_GEMSSNSTAL (SEQ ID NO:357) and A*01:01_EVDPIGHLY (SEQ ID NO:354) are shown.

[0191] Figure 84 The flow cytometry results for the HLA-peptide target A*03:01_GVHGGILNK (SEQ ID NO:355) are shown. Figure 84 "EVDPIGHVY" was also disclosed as SEQ ID NO:2.

[0192] Figure 85 A shows the total number of isolated CD8+ T cells for each HLA-peptide target across all tested donors. Figure 85 A discloses SEQ ID NO 73, 414, 322, 356, 355, 357, 2 and 354 in the order of their appearance.

[0193] Figure 85 B shows the frequency of isolated CD8+ T cells for each HLA-peptide target across all tested donors. Figure 85 B discloses SEQ ID NO 354, 322, 414, 355, 2, 357, 356 and 73 in the order of their appearance.

[0194] Figure 86 A describes the number of unique TCR clones for each HLA-peptide target from each donor for each test. Figure 86A discloses SEQ ID NO 73, 322, 356, 355, 357, 2 and 354 in the order of their appearance.

[0195] Figure 86 B depicts the total number of unique clones for each HLA-peptide target across all tested donors. Figure 86 B discloses SEQ ID NO 73, 322, 356, 355, 357, 2 and 354 in the order of their appearance.

[0196] Figure 87 Examples of Jurkat cells expressing A*0201_LLASSILCA (SEQ ID NO:322), A*0201_GVYDGEEHSV (SEQ ID NO:356), B*4402_GEMSSNSTAL (SEQ ID NO:357), and A*0101_EVDPIGHLY (SEQ ID NO:354)-specific TCRs are shown, which bind to their respective HLA-peptide targets but not to control peptide tetramers.

[0197] Figure 88 The gating strategy and flow data are shown, which demonstrate that human CD8+ cells transduced with the TCR identified in this paper bind to their specific HLA-peptide targets. Figure 88 "LLASSILCA" is disclosed as SEQ ID NO:322.

[0198] Figure 89 An exemplary lentiviral vector that can be used to transduce recipient cells using the TCR disclosed herein is shown.

[0199] Figure 90 This shows the BLI results for the G2 target Fab clone G2-P2C06. Figure 90 SEQ ID NO:73 has been disclosed.

[0200] Figure 91 A depicts the stability results of a second experiment from the G2 position variant HLA. Figure 91 A disclosed SEQ ID NO:73.

[0201] Figure 91 B depicts the binding affinity of Fab clone G2-P2C06 to the G2 position variant HLA. Figure 91 B disclosed SEQ ID NO:73.

[0202] Figure 92HDX heatmaps of the second round of HDX experiments are shown for the HLAα1 helix, HLAα2 helix, and the restriction peptide ASSLPTTMNY (SEQ ID NO:260) of G10 ABP from various tests. Figure 92 The amino acid sequence beginning with “KMEPR…” is also disclosed as SEQ ID NO:647 and the amino acid sequence beginning with “AADMA…” is disclosed as SEQ ID NO:648.

[0203] Figure 93 HDX heatmaps from the second round of HDX experiments are shown for the HLAα1 helix, HLAα2 helix, and the restriction peptide NTDNNLAVY (SEQ ID NO:73) from the tested G2 ABP. Figure 93 The amino acid sequence beginning with “KMEPR…” is also disclosed as SEQ ID NO:647 and the amino acid sequence beginning with “AADMA…” is disclosed as SEQ ID NO:648.

[0204] Figure 94 An example of HDX data from scFv G2-P2C11 plotted on crystal structure PDB 5bs0 is shown. Figure 94 "RRVY" is disclosed as SEQ ID NO:646.

[0205] Figure 95 High-resolution HDX data plotted on the PDB 5bs0 crystal structure are shown. As described in the experimental procedure, peptide fragmentation was performed via electron transfer dissociation (ETD) to obtain data for G2 binding to four different scFvs. Peptide fragments with high-resolution data (at approximately single amino acid resolution) and residues 157–160 are shown circled. Figure 95 "RRVY" is disclosed as SEQ ID NO:646. Figure 95 The amino acid sequence beginning with “TDRAN…” is also disclosed as SEQ ID NO:663 and the amino acid sequence beginning with “LRSWT…” is disclosed as SEQ ID NO:664.

[0206] Figure 96 Color thermograms of HDX experiments from all ABPs tested against the HLA-PEPTIDE target G5 (HLA-B*35:01_EVDPIGHVY(SEQ ID NO:2)) are shown. These include HLAα1 helices, HLAα2 helices, and the restriction peptide EVDPIGHVY(SEQ ID NO:2). Figure 96The amino acid sequence beginning with “RTEPR…” is also disclosed as SEQ ID NO:665 and the amino acid sequence beginning with “AADTA…” is disclosed as SEQ ID NO:649.

[0207] Figure 97 Showing Figure 96 The digital representation of a color heatmap. Figure 97 "EVDPIGHVY" is disclosed as SEQ ID NO:2. Figure 97 The amino acid sequence beginning with “RTEPR…” is also disclosed as SEQ ID NO:665 and the amino acid sequence beginning with “AADTA…” is disclosed as SEQ ID NO:649.

[0208] Figure 98 An example of data from the scFv clone G5-P1C12 plotted on the crystal structure of HLA-B*35:01 is shown (5xos.pdb; https: / / www.rcsb.org / structure / 5XOS).

[0209] Figure 99 Color thermograms of the second round of HDX experiments from all ABPs tested against the HLA-peptide target G8 (HLA-A*02:01_AIFPGAVPAA(SEQ ID NO:198)) are shown. Figure 99 The amino acid sequence beginning with “RMEPR…” is also disclosed as SEQ ID NO:643 and the amino acid sequence beginning with “AADMA…” is disclosed as SEQ ID NO:364.

[0210] Figure 100 Showing Figure 99 The digital representation of a color heatmap. Figure 100 The name "AIFPGAVPAA" is disclosed as SEQ ID NO:198. Figure 100 The amino acid sequence beginning with “RMEPR…” is also disclosed as SEQ ID NO:643 and the amino acid sequence beginning with “AADMA…” is disclosed as SEQ ID NO:364.

[0211] Figure 101 An example of high-resolution HDX data from scFv G8-P1H08 plotted on the crystal structure of Fab clone G8-P1C11 composite with HLA-PEPTIDE target A*02:01_AIFPGAVPAA (SEQ ID NO:198) (“G8”) is shown.

[0212] Figure 102 The results of flow cytometry experiments are shown, in which K562 cells transduced with HLA-B*35:01 were shocked with 50 μM of the target peptide EVDPIGHVY (SEQ ID NO:2) (“EVD”) or the negative control peptide IPSINVHHY (SEQ ID NO:358) (“IPS”), and pHLA-specific antibodies were detected by flow cytometry.

[0213] Figure 103 The results of flow cytometry experiments are shown, in which HLA-A*02:01 transduced K562 cells were shocked with 50 μM of the target peptide AIFPGAVPAA (SEQ ID NO:198) (“AIF”) or the negative control peptide FLLTRILTI (SEQ ID NO:359) (“FLL”), and pHLA-specific antibodies were detected by flow cytometry.

[0214] Figure 104 The results of flow cytometry experiments are shown, in which HLA-A*01:01 transduced K562 cells were shocked with 50 μM of the target peptide ASSLPTTMNY (SEQ ID NO:260) (“ASSL”) or the negative control peptide ATDALMTGY (SEQ ID NO:360) (“ATDA”), and pHLA-specific antibodies were detected by flow cytometry.

[0215] Figure 105 The results of BLI analysis of G8 target Fab clones G8-P4F05, G8-P1B03, and G8-P5G08 with HLA-peptide target A*02:01-AIFPGAVPAA (SEQ ID NO:198) are shown; as well as the results of BLI analysis of G5 target Fab clone G5-P1C12 with HLA-PEPTIDE target B*35:01-EVDPIGHVY (SEQ ID NO:2). Detailed Implementation

[0216] Unless otherwise defined, all technical terms, symbols, and other scientific terms used herein are intended to have the meanings commonly understood by those skilled in the art. In some instances, for clarity and / or ease of reference, terms are defined herein with their commonly understood meanings, and such definitions included herein are not necessarily construed as indicating a difference from the meanings commonly understood in the art. The methods and procedures described or referenced herein are methods and procedures commonly understood by those skilled in the art and typically applied using conventional methodologies, such as the widely used molecular cloning methods described, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual, 4th Edition (2012), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. Appropriately, unless otherwise stated, procedures regarding the use of commercially available kits and reagents are generally performed according to the manufacturer's defined protocols and conditions.

[0217] Unless the context clearly indicates otherwise, the singular forms “a,” “an,” and “the” used herein include the plural indicator. Unless otherwise clearly indicated, the terms “comprising,” “such as,” etc., are intended to convey a meaning that includes but is not limiting.

[0218] Unless otherwise specifically stated, the term "comprising" as used herein also specifically includes embodiments that "comprise the listed elements" and "comprise substantially the listed elements." For example, a multispecific ABP that "comprising a bifunctional antibody" includes a multispecific ABP that "comprises a bifunctional antibody" and a multispecific ABP that "comprises substantially a bifunctional antibody."

[0219] The term "about" refers to and covers both the indicated value and the range greater than and less than the value. In some embodiments, the term "about" means ±10%, ±5%, or ±1% of the specified value. In some embodiments, if applicable, the term "about" means the specified value ± one standard deviation of the value.

[0220] The term "immunoglobulin" refers to a class of structurally related proteins that typically consist of two pairs of polypeptide chains: a pair of light chains (L) and a pair of heavy chains (H). In a "complete immunoglobulin," all four chains are interconnected by disulfide bonds. The structure of immunoglobulins has been well characterized. See, for example, Paul, Fundamental Immunology, 7th Edition, Chapter 5 (2013), Lippincott, Williams & Wilkins, Philadelphia, PA. In short, each heavy chain typically contains a heavy chain variable region (V). H ) and heavy chain constant region (CH The heavy-chain constant region typically contains three structural domains, abbreviated as C. H1 C H2 and C H3 Each light chain typically contains a light chain variable region (V). L ( ) and the light chain constant region. The light chain constant region typically contains a structural domain, abbreviated as C. L .

[0221] The term “antigen-binding protein” or “ABP” as used herein is used in its broadest sense and includes certain types of molecules that contain one or more antigen-binding domains that specifically bind to an antigen or epitope.

[0222] In some embodiments, the ABP comprises an antibody. In some embodiments, the ABP is composed of an antibody. In some embodiments, the ABP is substantially composed of an antibody. Specifically, the ABP comprises an intact antibody (e.g., an intact immunoglobulin), an antibody fragment, an ABP fragment, and a multispecific antibody. In some embodiments, the ABP comprises an alternative scaffold. In some embodiments, the ABP is composed of an alternative scaffold. In some embodiments, the ABP is substantially composed of an alternative scaffold. In some embodiments, the ABP comprises an antibody fragment. In some embodiments, the ABP is substantially composed of an antibody fragment. In some embodiments, the ABP comprises a TCR or its antigen-binding portion. In some embodiments, the ABP is composed of a TCR or its antigen-binding portion. In some embodiments, the ABP is substantially composed of a TCR or its antigen-binding portion. In some embodiments, the CAR comprises the ABP. As provided herein, “HLA-peptide ABP,” “anti-HLA-peptide ABP,” or “HLA-peptide-specific ABP” is an ABP that specifically binds to the antigen HLA-peptide. ABP includes proteins containing one or more antigen-binding domains that specifically bind to antigens or epitopes via variable regions, such as variable regions derived from B cells (e.g., antibodies) or T cells (e.g., TCRs).

[0223] The term "antibody" in this article is used in its broadest sense and includes both polyclonal and monoclonal antibodies, encompassing complete antibodies and functional (antigen-binding) antibody fragments, including fragment antigen-binding (Fab) fragments, F(ab')2 fragments, Fab' fragments, Fv fragments, recombinant IgG (rIgG) fragments, and variable heavy chains (V) capable of specifically binding antigens. HThis term encompasses single-chain antibody fragments (including single-chain variable fragments (scFv)) and single-domain antibody fragments (e.g., sdAb, sdFv, nanobodies). The term covers genetically engineered and / or otherwise modified forms of immunoglobulins, such as intracellular antibodies, peptide bodies, chimeric antibodies, fully human antibodies, humanized antibodies and conjugated antibodies, multispecific antibodies (e.g., bispecific antibodies), biantibodies, triantibodies and tetraantibodies, tandem bivalent scFv, and tandem trivalent scFv. Unless otherwise stated, the term "antibody" should be understood to encompass its functional antibody fragment. The term also covers complete or full-length antibodies, including antibodies of any class or subclass, including IgG and its subclasses, IgM, IgE, IgA, and IgD.

[0224] As used in this article, “variable region” refers to a variable nucleotide sequence resulting from a recombination event, which may include, for example, the V, J, and / or D regions of an immunoglobulin or T cell receptor (TCR) sequence from a B cell or T cell (such as an activated T cell or an activated B cell).

[0225] The term "antigen-binding domain" refers to the portion of an ABP that can specifically bind to an antigen or epitope. An example of an antigen-binding domain is an antibody V against an ABP. H -V L The antigen-binding domain is formed by the dimer. Another example of an antigen-binding domain is an antigen-binding domain formed by diversifying certain loops from the tenth fibronectin type III domain of adnectin. The antigen-binding domain may sequentially contain antibody CDR1, CDR2, and CDR3 from the heavy chain; and antibody CDR1, CDR2, and CDR3 sequentially from the light chain. The antigen-binding domain may contain TCR CDRs, such as αCDR1, αCDR2, αCDR3, βCDR1, βCDR2, and βCDR3. TCR CDRs are described herein.

[0226] antibody V H District and V L The region can be further subdivided into highly variable regions (“HVR”; also known as “complementary determinant regions” (CDR)), interspersed with more conservative regions. The more conservative regions are called framing regions (FR). Each V H and V LTypically, it contains three antibody CDRs and four FRs, arranged in the following order (from N-terminus to C-terminus): FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. Antibody CDRs participate in antigen binding and influence antigen specificity and binding affinity of ABP. See Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed. (1991), Public Health Service, National Institutes of Health, Bethesda, MD, which is incorporated herein by reference in its entirety.

[0227] Based on the sequence of constant structural domains in vertebrates, light chains from any vertebrate can be divided into two types, called κ-kappa and λ-lambda.

[0228] Heavy chains in any vertebrate can be classified into one of five distinct classes (or isotypes): IgA, IgD, IgE, IgG, and IgM. These classes are also referred to as α, δ, ε, γ, and μ, respectively. Based on sequence and functional differences, IgG and IgA are further subdivided into subclasses. Humans express the following subclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

[0229] Those skilled in the art can use any of the many known numbering schemes to determine the amino acid sequence boundaries of antibody CDRs. Known numbering schemes include those described in the following literature: Kabat et al., above (“Kabat” numbering scheme); Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948 (“Chothia” numbering scheme); MacCallum et al., 1996, J. Mol. Biol., 262:732-745 (“Contact” numbering scheme); Lefran et al., Dev. Comp. Immunol., 2003, 27:55-77 (“IMGT” numbering scheme); and Honegge and Plückthun, J. Mol. Biol., 2001, 309:657-70 (“AHo” numbering scheme); each of which is incorporated herein by reference in its entirety.

[0230] Table 20 provides the positions of antibodies CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3 identified using the Kabat and Chothia schemes. For CDR-H1, residue numbers are provided using both the Kabat and Chothia numbering schemes.

[0231] For example, ABP numbering software (such as Abnum) can be used to assign antibody CDRs. Abnum is available from www.bioinf.org.uk / abs / abnum / and is described in Abhinandan and Martin, Immunology, 2008, 45:3832-3839 (which is incorporated in its entirety by reference).

[0232]

[0233] *When using the Kabat numbering convention, the C end of CDR-H1 varies between H32 and H34 depending on the length of the CDR.

[0234] When referring to residues in the constant region of the ABP heavy chain, the “EU numbering scheme” is generally used (e.g., as reported by Kabat et al., see above). Unless otherwise stated, the EU numbering scheme is used to refer to residues in the constant region of the ABP heavy chain as described herein.

[0235] The terms “full-length antibody,” “intact antibody,” and “whole antibody” used herein are interchangeable and refer to antibodies that have a structure substantially similar to that of naturally occurring antibodies and that contain a heavy chain including an Fc region. For example, when used to refer to IgG molecules, a “full-length antibody” is an antibody that consists of two heavy chains and two light chains.

[0236] Those skilled in the art can use any of the many known numbering schemes to determine the amino acid sequence boundaries of TCR CDRs, including but not limited to the unique IMGT numbering described in the following references: LeFranc, M.-P, Immunol Today. November 1997; 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 Facts Book; and M.-P. Lefranc / Developmental and Comparative Immunology 27 (2003) 55–77; all of which are incorporated herein by reference.

[0237] "ABP fragment" includes a portion of the complete ABP, such as the antigen-binding or variable region of the complete ABP. ABP fragments include, for example, Fv fragments, Fab fragments, F(ab')2 fragments, Fab' fragments, scFv((sFv)) fragments, and scFv-Fc fragments. ABP fragments also include antibody fragments. Antibody fragments can include Fv fragments, Fab fragments, F(ab')2 fragments, Fab' fragments, scFv((sFv)) fragments, scFv-Fc fragments, and TCR fragments.

[0238] The “Fv” fragment is a non-covalently linked dimer consisting of a heavy chain variable domain and a light chain variable domain.

[0239] In addition to the variable domains of the heavy chain and the light chain, the "Fab" fragment also contains a constant domain of the light chain and a first constant domain ((CH1)) of the heavy chain. The Fab fragment can be produced, for example, by recombinant methods or by papain digestion of a full-length ABP.

[0240] The “F(ab’)2” fragment contains two Fab’ fragments linked by disulfide bonds near the hinge region. The F(ab’)2 fragment can be produced, for example, by recombinant methods or by pepsin digestion of the intact ABP. The F(ab’) fragment can be dissociated, for example, by treatment with β-mercaptoethanol.

[0241] The "single-chain Fv" or "sFv" or "scFv" fragment contains a VH domain and a VL domain within a single polypeptide chain. The VH and VL domains are typically linked by a peptide linker. See Plückthun A. (1994). Any suitable linker can be used. In some embodiments, the linker is (GGGGS). n (SEQ ID NO:361). In some implementations, n = 1, 2, 3, 4, 5, or 6. See ABP derived from Escherichia coli. Rosenberg M. & Moore GP (eds.), The Pharmacology of Monoclonal ABPs, Vol. 113 (pp. 269–315). Springer-Verlag, New York, which is incorporated herein by reference in its entirety.

[0242] The “scFv-Fc” fragment contains an scFv attached to an Fc domain. For example, the Fc domain may be attached to the C-terminus of the scFv. Depending on the orientation of the variable domains in the scFv (i.e., VH-VL or VL-VH), the Fc domain may be after VH or VL. Any suitable Fc domain known in the art or described herein may be used. In some cases, the Fc domain includes the IgG4Fc domain.

[0243] The term "single-domain antibody" refers to a molecule in which one variable domain of an ABP specifically binds to an antigen while the other variable domain is absent. Single-domain ABPs and fragments thereof are described in: Arabi Ghahroudi et al., FEBS Letters, 1998, 414:521-526 and Muyldermans et al., Trends in Biochem. Sci., 2001, 26:230-245, each incorporated herein by reference in its entirety. Single-domain ABPs are also known as sdAbs or nanobodies.

[0244] The term "Fc region" or "Fc" refers to the C-terminal region of the immunoglobulin heavy chain, which interacts with Fc receptors and certain proteins of the complement system in naturally occurring antibodies. The structure of the Fc regions of various immunoglobulins and the glycosylation sites contained therein are known in the art. See Schroeder and Cavacini, J. Allergy Clin. Immunol., 2010, 125:S41-52, which are incorporated herein by reference in their entirety. The Fc region can be a naturally occurring Fc region or a modified Fc region as described in the art or elsewhere in this disclosure.

[0245] 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 domains bind to the antigen or epitope with a specificity and affinity similar to that of ABP. Exemplary alternative scaffolds include those derived from fibronectin (e.g., Adnectins). TM ), β-sandwich (e.g., iMab), lipid transport proteins (e.g., ), EETI-II / AGRP, BPTI / LACI-D1 / ITI-D2 (e.g., Kunitz domain), thioredoxin peptide aptamers, protein A (e.g., These include ankyrin repeat sequences (e.g., DARPins), γ-β-crystallins / ubiquitous proteins (e.g., Affilins), CTLD3 (e.g., Tetranectins), Fynomers, and (LDLR-A modules) (e.g., Avimers). Further information on alternative scaffolds is provided in the following literature: 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 herein by reference in its entirety. One alternative scaffold is a type of ABP.

[0246] A "multispecific ABP" is an ABP comprising two or more distinct antigen-binding domains that collectively and specifically bind to two or more distinct epitopes. The two or more distinct epitopes can be epitopes on the same antigen (e.g., a single HLA-peptide molecule expressed by a cell) or epitopes on different antigens (e.g., different HLA-peptide molecules expressed by the same cell, or HLA-peptide and non-HLA-peptide molecules). In some aspects, a multispecific ABP binds to two distinct epitopes (i.e., a "bispecific ABP"). In some aspects, a multispecific ABP binds to three distinct epitopes (i.e., a "trispecific ABP").

[0247] A "monospecific ABP" is an ABP that contains one or more binding sites that specifically bind to a single epitope. An example of a monospecific ABP is the naturally occurring IgG molecule, which, although bivalent (i.e., having two antigen-binding domains), recognizes the same epitope on each of the two antigen-binding domains. Binding specificity can exist at any suitable valence.

[0248] The term "monoclonal antibody" refers to an antibody derived from a substantially homogeneous population of antibodies. A substantially homogeneous population of antibodies contains broadly similar antibodies that bind to the same epitopes, except for variants that may typically occur during monoclonal antibody production. Typically, only a small number of such variants are present. Monoclonal antibodies are usually obtained by methods involving the selection of a single antibody from a plurality of antibodies. For example, the selection method may be to choose a unique clone from a plurality of clones, such as a hybridoma clone, a phage clone, a yeast clone, a bacterial clone, or a collection of other recombinant DNA clones. The selected antibody may be further modified, for example, to improve affinity for the target ("affinity maturation"), to humanize the antibody, to improve its production in cell cultures, and / or to reduce its immunogenicity in subjects.

[0249] The term "chimeric antibody" refers to an antibody in which a portion of the heavy chain and / or light chain originates from a specific source or species, while the remainder of the heavy chain and / or light chain originates from a different source or species.

[0250] A “humanized” form of a nonhuman antibody is a chimeric antibody containing a minimal sequence derived from a nonhuman antibody. A humanized antibody is typically a human antibody (recipient antibody) in which one or more CDR residues are replaced by residues of one or more CDRs of a nonhuman antibody (donor antibody). The donor antibody can be any suitable nonhuman antibody, such as mouse, rat, rabbit, chicken, or nonhuman primate antibodies with the desired specificity, affinity, or biological effect. In some cases, selected frame region residues of the recipient antibody are replaced by corresponding frame region residues of the donor antibody. Humanized antibodies may also contain residues not found in either the recipient or donor antibody. This modification can be performed to further improve antibody function. For further details, see Jones et al., Nature, 1986, 321:522-525; Riechmann et al., Nature, 1988, 332:323-329; and Presta, Curr. Op. Struct. Biol., 1992, 2:593-596, each of which is incorporated in its entirety by reference.

[0251] "Human antibody" is an antibody having an amino acid sequence that corresponds to the amino acid sequence of an antibody produced by a human or human cell, or derived from a non-human source (e.g., obtained from a human source or designed de novo) using a human antibody library or a human antibody encoding sequence. Human antibodies specifically exclude humanized antibodies.

[0252] “Affinity” refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., ABP) and its binding mate (e.g., an antigen or epitope). Unless otherwise stated, “affinity” as used herein refers to intrinsic binding affinity, which reflects a 1:1 interaction between members of the binding pair (e.g., ABP and an antigen or epitope). The affinity of molecule X for its mate Y can be expressed using the dissociation equilibrium constant (K0). D The following describes the kinetic elements relating to the dissociation equilibrium constant in more detail. Affinity can be measured by conventional methods known in the art, including those described herein, such as surface plasmon resonance (SPR) techniques (e.g., ) or biological layer interferometry (e.g., ).

[0253] Regarding the binding of ABP to target molecules, the terms "binding to a specific antigen (e.g., a peptide target) or an epitope on a specific antigen," "specifically binding to a specific antigen (e.g., a peptide target) or an epitope on a specific antigen," "specifically binding to a specific antigen (e.g., a peptide target) or an epitope on a specific antigen," "specific to a specific antigen (e.g., a peptide target) or an epitope on a specific antigen," "selectively binding to a specific antigen (e.g., a peptide target) or an epitope on a specific antigen," and "selectively binding to a specific antigen (e.g., a peptide target) or an epitope on a specific antigen" refer to binding that is distinctly different from nonspecific or nonselective interactions (e.g., binding to non-target molecules). Specific binding can be measured, for example, by measuring the binding to a target molecule and comparing it to the binding to a non-target molecule. Specific binding can also be determined by competition with a control molecule that mimics the epitope recognized on the target molecule. In that case, if the control molecule competitively inhibits the binding of ABP to the target molecule, it indicates specific binding. In some respects, HLA-peptide ABP has an affinity for non-target molecules that is approximately 50% less than its affinity for HLA-peptides. In some respects, HLA-peptide ABP has an affinity for non-target molecules that is approximately 40% less than its affinity for HLA-peptides. In some respects, HLA-peptide ABP has an affinity for non-target molecules that is approximately 30% less than its affinity for HLA-peptides. In some respects, HLA-peptide ABP has an affinity for non-target molecules that is approximately 20% less than its affinity for HLA-peptides. In some respects, HLA-peptide ABP has an affinity for non-target molecules that is approximately 10% less than its affinity for HLA-peptides. In some respects, HLA-peptide ABP has an affinity for non-target molecules that is approximately 1% less than its affinity for HLA-peptides. In some respects, HLA-peptide ABP has an affinity for non-target molecules that is approximately 0.1% less than its affinity for HLA-peptides.

[0254] The term "k" used in this article d (sec) -1 () refers to the dissociation rate constant of a specific ABP-antigen interaction. This value is also known as k. off value.

[0255] The term "k" used in this article a (M) -1 ×sec -1 () refers to the association rate constant of a specific ABP-antigen interaction. This value is also known as k. on value.

[0256] The term "K" used in this article D (M) refers to the dissociation equilibrium constant of a specific ABP-antigen interaction. K D =k d / k aIn some implementations, the affinity of ABP is based on the K-axis against the interaction between the ABP and its antigen. D Described. For clarity, as is known in the art, the smaller K... D A higher K value indicates a stronger affinity interaction, while a larger K value indicates a weaker affinity interaction. D The value indicates a lower affinity interaction.

[0257] The term "K" used in this article A (M) -1 K refers to the association equilibrium constant of a specific ABP-antigen interaction. A =k a / k d .

[0258] "Immune conjugate" is an ABP conjugated with one or more heterologous molecules, such as therapeutic agents (e.g., cytokines) or diagnostic agents.

[0259] "Fc effector function" refers to biological activities mediated by the Fc region of an ABP containing an Fc region, and these activities can vary by subtype. Examples of ABP effector functions include C1q binding to activate complement-dependent cytotoxicity (CDC), Fc receptor binding to activate ABP-dependent cytotoxicity (ADCC), and ABP-dependent phagocytosis (ADCP).

[0260] When used in the context of two or more ABPs, the terms "competing with" or "cross-competing with" mean that two or more ABPs compete to bind to an antigen (e.g., an HLA-peptide). In one exemplary assay, an HLA-peptide is coated onto a surface and brought into contact with a first HLA-peptide ABP, and then a second HLA-peptide ABP is added. In another exemplary assay, a first HLA-peptide ABP is coated onto a surface and brought into contact with an HLA-peptide, and then a second HLA-peptide ABP is added. If, in either assay, 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 "competing with" also includes combinations of ABPs where one ABP reduces the binding of another ABP, but no competition is observed when the ABPs are added in reverse order. However, in some embodiments, the first and second ABPs inhibit each other's binding regardless of their order of addition. 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%. A technician can select the concentration of the ABP used for the competitive assay based on the ABP's affinity for the HLA-peptide and the ABP's valence. The assays described in this definition are illustrative, and a technician can use any suitable assay to determine whether ABPs compete with each other. The appropriate assays are described in the following references: for example, Cox et al., “Immunoassay Methods” in the AssayGuidance Manual updated December 24, 2014 [Internet] (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; each of which is incorporated herein by reference in its entirety.

[0261] The term "epitope" refers to a portion of an antigen that specifically binds to ABP. Epitopes typically consist of surface-accessible amino acid residues and / or sugar side chains and may possess specific three-dimensional structural characteristics and specific charge properties. The difference between conformational and non-conformational epitopes is that binding to the former, but not the latter, may be lost in the presence of denaturing solvents. Epitopes may contain amino acid residues that directly participate in binding and other amino acid residues that do not directly participate in binding. Known techniques for identifying epitopes can be used to determine epitopes that bind to ABP, such as, for example, testing the binding of ABP to HLA-peptide variants with different point mutations or to chimeric HLA-peptide variants.

[0262] The percentage of “identity” between a polypeptide sequence and a reference sequence is defined as the percentage of amino acid residues in the polypeptide sequence that are identical to those in the reference sequence after sequence alignment and the introduction of gaps (if necessary) to obtain the maximum percentage of sequence identity. Alignment used to determine the percentage of amino acid sequence identity can be performed in various ways within the scope of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, MEGALIGN (DNASTAR), CLUSTALW, CLUSTAL OMEGA, or MUSCLE software. Those skilled in the art can determine suitable parameters for sequence alignment, including any algorithm required to achieve maximum alignment across the full length of the sequences being compared.

[0263] "Conservative substitution" or "conservative amino acid substitution" refers to the substitution of one amino acid by another amino acid that is chemically or functionally similar to it. Conservative substitution of similar amino acids is well known in the art. For example, in some embodiments, the amino acid groups provided in Tables 21-23 are considered to be conservative substitutions of each other.

[0264] Table 21. Selected amino acid groups that are considered to be conservatively substituted for each other in some embodiments.

[0265] acidic residues D and E basic residues K, R, and H Hydrophilic uncharged residues S, T, N, and Q Aliphatic uncharged residues G, A, V, L and I Nonpolar uncharged residues C, M, and P Aromatic residues F, Y and W

[0266] Table 22. In some embodiments, other selected amino acid groups are considered to be conservatively substituted for each other.

[0267] Group 1 A, S, and T Group 2 D and E Group 3 N and Q Group 4 R and K Group 5 I, L and M Group 6 F, Y and W

[0268] Table 23. In some embodiments, a further selected group of amino acids that are considered to be conservatively substituted for each other.

[0269] Group A A and G Group B D and E Group C N and Q Group D R, K and H Group E I, L, M, V Group F F, Y and W Group G S and T Group H C and M

[0270] Other conserved substitutions can be found, for example, in Creighton, Proteins: Structures and Molecular Properties, 2nd Edition (1993), WH Freeman & Co., New York, NY. ABPs produced by making one or more conserved substitutions to amino acid residues in the parental ABP are called “conservatively modified variants”.

[0271] The term "amino acid" refers to twenty common, naturally occurring amino acids. These include 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).

[0272] As used herein, the term "vector" refers to a nucleic acid molecule capable of replicating another nucleic acid linked to it. This term includes vectors that function as self-replicating nucleic acid structures, as well as vectors incorporated into the host cell genome. Some vectors are capable of directing the expression of nucleic acids operatively linked to them. Such vectors are referred to herein as "expression vectors."

[0273] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acids have been introduced and their progeny. Host cells include "transformants" (or "transformed cells") and "transfectants" (or "transfected cells"), each comprising primary transformed or transfected cells and their derived progeny. These progeny may not be identical to the parent cells in terms of nucleic acid content and may contain mutations.

[0274] The term "treating" (and its variations, such as "treat" or "treatment") refers to a clinical intervention that attempts to alter the natural course of a disease or symptom in a subject in need. Treatment can be aimed at prevention and at addressing clinicopathological processes. Ideal therapeutic outcomes include preventing the onset or recurrence of disease, alleviating symptoms, reducing any direct or indirect pathological consequences of the disease, preventing metastasis, slowing the rate of disease progression, improving or mitigating the disease state, and alleviating or improving prognosis.

[0275] As used herein, the term "therapeutic effective amount" or "effective amount" refers to the amount of ABP or pharmaceutical composition provided herein that, when administered to a subject, is effective in treating a disease or condition.

[0276] As used herein, the term "subject" refers to a mammalian subject. Exemplary subjects include humans, monkeys, dogs, cats, mice, rats, cattle, horses, camels, goats, rabbits, and sheep. In some embodiments, the subject is a human. In some embodiments, the subject suffers from a disease or condition that can be treated with the ABP provided herein. In some aspects, the disease or condition is cancer. In some aspects, the disease or condition is a viral infection.

[0277] The term "instructions for use" is used to refer to instructions typically included in the commercial packaging of a therapeutic or diagnostic product (e.g., a test kit), which contain information about the indications, usage, dosage, administration, combination therapy, contraindications, and / or warnings for using such a therapeutic or diagnostic product.

[0278] The term "tumor" refers to the growth and proliferation of all proliferative cells (whether malignant or benign), as well as all precancerous and cancerous cells and tissues. The terms "cancer," "carcinoma," "cellular proliferative disorder," "proliferative lesion," and "tumor" are not mutually exclusive herein. The terms "cellular proliferative disorder" and "proliferative lesion" refer to conditions associated with some degree of abnormal cell proliferation. In some implementations, a cellular proliferative disorder is cancer. In some respects, a tumor is a solid tumor. In some respects, a tumor is a hematologic malignancy.

[0279] The term "pharmaceutical composition" refers to a formulation which exists in a form that allows the biological activity of the active ingredient contained therein to effectively treat a subject, and which does not contain any additional components provided in the pharmaceutical composition that would have unacceptable toxicity to the subject.

[0280] The terms “modulate” and “modulation” refer to reducing or suppressing, or alternatively, activating or increasing, the listed variables.

[0281] The terms “increase” and “activation” refer to an increase of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 2 times, 3 times, 4 times, 5 times, 10 times, 20 times, 50 times, 100 times or more in the listed variables.

[0282] The terms “reduction” and “suppression” refer to a reduction of the listed variables by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 2 times, 3 times, 4 times, 5 times, 10 times, 20 times, 50 times, 100 times or more.

[0283] The term "agonist" refers to the activation of receptor signaling to induce a biological response associated with receptor activation. An "agonist" is an entity that binds to and activates receptors.

[0284] The term "antagonist" refers to the inhibition of receptor signaling to suppress biological responses associated with receptor activation. An "antagonist" is an entity that binds to and antagonizes a receptor.

[0285] The terms "nucleic acid" and "polynucleotide" are used interchangeably in this document and refer to a polymer of nucleotides of any length, namely deoxyribonucleotides or ribonucleotides or their analogues. Polynucleotides may include, but are not limited to, coding or non-coding regions of genes or gene fragments, loci as defined from the perspective of linkage analysis, exons, introns, messenger RNA (mRNA), cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA, isolated RNA, nucleic acid probes, and primers. Polynucleotides may include modified nucleotides, such as methylated nucleotides and nucleotide analogues. 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-galactosylinosine, inosine, N6-isopentene adenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-substituted adenine, 7-methylguanine. 5-Methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, β-D-mannosyl queosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthioN6-isopentene adenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, methyl uracil-5-oxyacetic acid, 3-(3-amino-3-N-2-carboxypropyl)uracil, and 2,6-diaminopurine.

[0286] Isolated HLA-peptide targets

[0287] The major histocompatibility complex (MHC) is a complex of antigens encoded by a set of linked loci. These antigens are collectively referred to as H-2 in mice and HLA in humans. MHC antigens have two main classes, class I and class II, each comprising a group of cell surface glycoproteins that play a role in determining tissue type and transplant compatibility. In transplant response, cytotoxic T cells (CTLs) primarily respond to class I glycoproteins, while helper T cells primarily respond to class II glycoproteins.

[0288] Human major histocompatibility complex (MHC) class I molecules (which may be interchangeably referred to herein as HLA class I molecules) are expressed on the surface of almost all cells. These molecules function to present peptides, primarily derived from endogenously synthesized proteins, to cells such as CD8+ T cells through interaction with α-β T cell receptors. MHC class I molecules consist of a heterodimer consisting of a 46 kDa α chain that is non-covalently associated with a 12 kDa light chain β-2 microglobulin. The α chain typically contains α1 and α2 domains that form the groove for presenting HLA-restricted peptides, and an α3 transmembrane domain that interacts with the CD8 co-receptor on T cells. Figure 1 (Prior art) describes the general structure of HLA class I molecules. Some TCRs can bind to MHC class I molecules 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).

[0289] Class I MHC-restricted peptides (which may also be referred to interchangeably herein as HLA-restricted antigens, HLA-restricted peptides, MHC-restricted antigens, restricted peptides, or peptides) generally bind to the α1-α2 groove of the heavy chain via about two or three anchoring residues that interact with the corresponding binding pockets in the MHC molecule. The β-2-microglobulin chain plays a crucial role in intracellular transport, peptide binding, and conformational stability of MHC class I molecules. 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 β-2-microglobulin leads to protein maturation and export to the cell surface.

[0290] The binding of a given HLA subtype to an HLA-restricted peptide forms a complex with a unique and novel surface, which can be specifically recognized by ABPs (e.g., TCRs on T cells or their antibody or antigen-binding fragments). The HLA complexed with the HLA-restricted peptide is referred to herein as an HLA-peptide, pHLA, or HLA-peptide target. In some cases, the restriction peptide is located in the α1 / α2 groove of the HLA molecule. In other cases, the restriction peptide binds to the α1 / α2 groove of the HLA molecule via about two or three anchoring residues that interact with the corresponding binding pocket in the HLA molecule.

[0291] Therefore, this article provides antigens containing HLA-peptide targets. HLA-peptide targets may contain specific HLA-restricted peptides with a defined amino acid sequence that complexes with a specific HLA subtype.

[0292] The HLA-peptide targets identified in this paper can be used for cancer immunotherapy. In some embodiments, the HLA-peptide targets identified in this paper are present on the surface of tumor cells. The HLA-peptide targets identified in this paper can be expressed by tumor cells in human subjects. The HLA-peptide targets identified in this paper can be expressed by tumor cells in a human subject population. For example, the HLA-peptide targets identified in this paper can be common antigens that are typically expressed in human subject populations with cancer.

[0293] The HLA-peptide targets identified in this study exhibit varying prevalence rates depending on the individual tumor type. These prevalence rates 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%, 4... 4%, 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%, or 100%. The prevalence varies depending on the individual tumor type, ranging from approximately 0.1% to 100%, 0.2% to 50%, 0.5% to 25%, or 1% to 10%.

[0294] Preferably, HLA-peptide targets are not typically expressed in most normal tissues. For example, in some cases, HLA-peptide targets may not be expressed in tissues included in a Genotype Tissue Expression (GTEx) project, or in some cases, they may only be expressed in immune-privileged or non-essential tissues. Exemplary immune-privileged or non-essential tissues include the testes, minor salivary glands, cervical endometrium, and thyroid gland. In some cases, an HLA-peptide target may be considered not expressed in essential or non-immune-privileged tissues if the median expression of the gene for which the restriction peptide is derived is less than 0.5 RPKM (number of reads per kilobase transcript per million mapped reads) in a GTEx sample, if the gene expression in a GTEx sample does not exceed 10 RPKM, if the gene is expressed at ≥5 RPKM in no more than two samples across all essential tissue samples, or any combination thereof.

[0295] Exemplary HLA class I subtypes of HLA-peptide targets

[0296] Anyone who is a member of the MHC (not a member of the MHC) HL She is also an MHC member of HLA) and she is not HLA member (also HLA-A2、HLA-A1、HLA-A3、HLA-A11、HLA-A23、HLA-A24、HLA-A25、HLA- A26、HLA-A28、HLA-A29、HLA-A30、HLA-A31、HLA-A32、HLA-A33、HLA-A34、HL A-68、PAGE-B7、PAGE-B8、PAGE-B40、PAGE-B44、PAGE-B13、PAGE-B15、PAGE-B-18、PAGE -B27、HLA-B35、HLA-B37、HLA-B38、HLA-B39、HLA-B45、HLA-B46、HLA-B49、H LA-B51、HLA-B54、HLA-B55、HLA-B56、HLA-B57、HLA-B58、HLA-C*01、HLA-C* 02、HLA-C*03、HLA-C*04、HLA-C*05、HLA-C*06、HLA-C*07、HLA-C*12、HLA-C *14、HLA-C*16、HLA-Cw8、HLA-A*01:01、HLA-A*02:01、HLA-A*02:03、HLA-A* 02:04、HLA-A*02:07、HLA-A*03:01、HLA-A*03:02、HLA-A*11:01、HLA-A*23 :01、HLA-A*24:02、HLA-A*25:01、HLA-A*26:01、HLA-A*29:02、HLA-A*30:0 1、HLA-A*30:02、HLA-A*31:01、HLA-A*32:01、HLA-A*33:01、HLA-A*33:03、 HLA-A*68:01、HLA-A*68:02、HLA-B*07:02、HLA-B*08:01、HLA-B*13:02、HLA -B*15:01、HLA-B*15:03、HLA-B*18:01、HLA-B*27:02、HLA-B*27:05、HLA-B *35:01、HLA-B*35:03、HLA-B*37:01、HLA-B*38:01、HLA-B*39:01、HLA-B*4 0:01、HLA-B*40:02、HLA-B*44:02、HLA-B*44:03、HLA-B*46:01、HLA-B*49: 01、HLA-B*51:01、HLA-B*54:01、HLA-B*55:01、HLA-B*56:01、HLA-B*57:01、HLA-B*58:01, HLA-C*01:02, HLA-C*02:02, HLA-C*03:03, HLA-C*03:04, HLA-C*04:01, HLA-C*05:01, HLA-C*06:02, HLA-C*07:01, HLA-C*07:02, HLA-C*07:04, HLA-C*07:06, HLA-C*12:03, HLA-C*14:02, HLA-C*16:01, HLA-C*16:02, HLA-C*16:04, and all their subtypes, including, for example, 4-position, 6-position, and 8-position subtypes. Allelic variants of the above-described HLA types exist, and this invention covers all such allelic variants. 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.

[0297] HLA-restricted peptide

[0298] HLA-restricted peptides (which may be referred to interchangeably herein) "restricted peptides" can be peptide fragments of tumor-specific genes (e.g., cancer-specific genes). Preferably, the cancer-specific gene is expressed in a cancer sample. Genes aberrantly expressed in cancer samples can be identified using databases. Exemplary databases, illustrated only by way of enumeration, include: The Cancer Genome Atlas (TCGA) Research Network: http: / / cancergenome.nih.gov / ; International Cancer Genome Association: https: / / dcc.icgc.org / . In some embodiments, the cancer-specific gene is observed to have at least 10 RPKM of expression in at least 5 samples from the TCGA database. The cancer-specific gene may have an observable bimodal distribution.

[0299] The cancer-specific gene may have observed expression levels greater than 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 transcripts per million (TPM) in at least one TCGA tumor tissue. In a preferred embodiment, the cancer-specific gene has observed expression levels greater than 100 TPM in at least one TCGA tumor tissue. In some cases, the cancer-specific gene exhibits an observed bimodal expression distribution in TCGA samples. Where it is not desired to be bound by theory, this bimodal expression pattern is consistent with biological models in which expression levels are minimum at baseline across all tumor samples, with higher expression levels in a subset of tumors that have undergone epigenetic dysregulation.

[0300] Preferably, cancer-specific genes are not typically expressed in most normal tissues. For example, in some cases, cancer-specific genes may not be expressed in tissues included in a Genotype Tissue Expression (GTEx) project, or in some cases, they may be expressed in immune-privileged or non-essential tissues. Exemplary immune-privileged or non-essential tissues include the testes, minor salivary glands, cervical endometrium, and thyroid gland. In some cases, a cancer-specific gene may be considered not expressed in an essential or non-immune-privileged tissue if the median expression of the cancer-specific gene in a GTEx sample is less than 0.5 RPKM (number of reads per kilobase transcript per million mapped reads), if the gene expression in a GTEx sample does not exceed 10 RPKM, if the gene is expressed at ≥5 RPKM in no more than two of all essential tissue samples, or any combination thereof.

[0301] In some implementations, cancer-specific genes are evaluated to meet the following criteria: (1) median GTEx expression in the brain, heart or lung is less than 0.1 transcripts per million (TPM), with no sample exceeding 5 TPM; (2) median GTEx expression in other essential organs (excluding the testes, thyroid gland and minor salivary glands) is less than 2 TPM, with no sample exceeding 10 TPM.

[0302] In some implementations, cancer-specific genes are generally unlikely to be expressed in immune cells; for example, they are not interferon family genes, not eye-related genes, not olfactory or gustatory receptor genes, and not genes associated with circadian rhythms (e.g., not CLOCK, PERIOD, CRY genes).

[0303] The restricted peptide is preferably present on the surface of the tumor.

[0304] The size of the restricted peptide can be 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, and any range that can be derived therefrom. In a particular embodiment, the size of the restricted peptide is about 8, about 9, about 10, about 11, or about 12 amino acid residues. The length of the restricted peptide can be about 5 to 15 amino acids, preferably about 7 to 12 amino acids, or more preferably about 8 to 11 amino acids.

[0305] Exemplary HLA-peptide targets

[0306] Exemplary HLA-peptide targets are shown in Tables A, A1, and A2. Each row in Tables A, A1, and A2 shows the HLA allele and the corresponding HLA-restricted peptide sequence for each complex. The peptide sequence may consist of the corresponding sequence shown in any row of Tables A, A1, or A2. Alternatively, the peptide sequence may comprise the corresponding sequence shown in any row of Tables A, A1, or A2. Alternatively, the peptide sequence may consist substantially of the corresponding sequence shown in any row of Tables A, A1, or A2.

[0307] In some implementations, the HLA-peptide target is the target shown in Table A, Table A1, or Table A2.

[0308] In some implementations, the HLA-peptide target is the target shown in Table A, Table A1, or Table A2, provided that the isolated HLA-peptide target is not any of target numbers 6364-6369, 6386-6389, 6500, 6521-6524, or 6578, and is not an HLA-peptide target present in Table B or Table C.

[0309] In some implementations, the HLA-restricted peptide is not derived from a gene selected from WT1 or MART1.

[0310] HLA class I molecules that do not associate with restriction peptide ligands are generally unstable. Therefore, association of the restriction peptide with the α1 / α2 groove of the HLA molecule can stabilize the non-covalent association between the β2-microglobulin subunit of the HLA isotype and the α-subunit of the HLA isotype.

[0311] The stability of the nonvalent association between the β2-microglobulin subunit of the HLA isotype and the α-subunit of the HLA isotype can be determined using any suitable method. For example, this stability can be assessed by dissolving the insoluble aggregates of the HLA molecule in a high concentration of urea (e.g., about 8 M urea) and determining the ability of the HLA molecule to refold in the presence of the restriction peptide during urea removal (e.g., by dialysis). This refolding method is described, for example, in Proc. Natl. Acad. Sci. USA, Vol. 89, pp. 3429-3433, April 1992, which is hereby incorporated by reference.

[0312] For other examples, conditional HLA class I ligands can be used to assess this stability. Conditional HLA class I ligands are typically designed as short restriction peptides that stabilize the association between the β2 and α subunits of an HLA class I molecule by binding to the α1 / α2 groove, and contain one or more amino acid modifications that cause the restriction peptide to cleave upon exposure to a conditional stimulus. Once the conditional ligand cleaves, the β2 and α subunits of the HLA molecule dissociate unless the conditional ligand is replaced with a restriction 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 HLA peptide ligands or predicted high-affinity HLA peptide ligands. For HLA alleles with available structural information, the water accessibility of the side chains can also be used to select the site for introducing amino acid modifications. The use of conditional HLA ligands may be advantageous by allowing the batch production of stable HLA-peptide complexes that can be used to query and test restriction peptides in a high-throughput manner.Conditional HLA class I ligands and their production methods are described in, for example, Proc Natl Acad Sci U SA. 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 Characterisation. 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 in their entirety by citation.

[0313] Therefore, in some embodiments, the ability of the HLA-restricted peptides described herein (e.g., those described in Table A, Table A1, or Table A2) to stabilize the association of the β2- and α-subunits of an HLA molecule is assessed by performing conditional ligand-mediated exchange reactions and HLA stability assays. HLA stability can be determined using any suitable method, including, for example, mass spectrometry, immunoassays (e.g., ELISA), size exclusion chromatography, and HLA multimer staining, followed by flow cytometry evaluation of T cells.

[0314] Other exemplary methods for assessing the stability of the non-covalent association between the β2-microglobulin subunit of an HLA isotype and the α-subunit of an HLA isotype include peptide exchange using dipeptides. Peptide exchange using dipeptides is described, for example, in ProcNatl Acad Sci US A. 2013 Sep 17; 110(38):15383-8; Proc Natl Acad Sci U SA. 2015 Jan 6; 112(1):202-7, which are incorporated herein by reference.

[0315] This article provides useful antigens containing HLA-peptide targets. HLA-peptide targets can contain specific HLA-restricted peptides with well-defined amino acid sequences that are complexed with specific HLA subtype alleles.

[0316] HLA-peptide targets can be isolated and / or in substantially pure form. For example, HLA-peptide targets can be isolated from their natural environment or produced by technical methods. In some cases, HLA-peptide targets are provided in a form that is substantially free of other peptides or proteins.

[0317] HLA-peptide targets may exist in soluble form and, optionally, may be recombinant HLA-peptide target complexes. Those skilled in the art can use any suitable method to generate and purify recombinant HLA-peptide targets. Suitable methods include, for example, using Escherichia coli expression systems, insect cells, etc. Other methods include synthetic generation, for example using cell-free systems. WO2017089756 describes exemplary suitable cell-free systems, which are hereby incorporated in their entirety by reference.

[0318] This article also provides compositions containing HLA-peptide targets.

[0319] In some cases, the composition comprises an HLA-peptide target attached to a solid support. Exemplary solid supports include, but are not limited to, beads, pores, membranes, tubes, columns, plates, agarose gels, magnetic beads, and fragments. Exemplary solid supports are described, for example, in Catalysts 2018, 8, 92; doi:10.3390 / catal8020092, which is hereby incorporated herein by reference in its entirety.

[0320] HLA-peptide targets can be attached to solid supports using any suitable method known in the art. In some cases, HLA-peptide targets are covalently attached to solid supports.

[0321] In some cases, HLA-peptide targets are attached to solid supports via affinity binding pairs. Affinity binding pairs typically involve specific interactions between two molecules. Ligands with affinity for their binding chaperone molecules can be covalently attached to solid supports and thus used as decoys to immobilize common affinity binding pairs, including, for example, streptavidin and biotin, avidin and biotin; and multihistidine tags with metal ions (such as copper, nickel, zinc, and cobalt).

[0322] HLA-peptide targets can contain detectable markers.

[0323] A pharmaceutical composition containing an HLA-peptide target.

[0324] Compositions containing HLA-peptide targets can be pharmaceutical compositions. Such compositions may contain multiple HLA-peptide targets. Exemplary pharmaceutical compositions are described herein. These compositions may be able to elicit an immune response. The compositions may contain adjuvants. Suitable adjuvants include, but are not limited to: 1018ISS, alum, aluminum salts, Amplivax, AS15, BCG, CP-870893, CpG7909, CyaA, dSLIM, GM-CSF, IC30, IC31, imiquimod, ImuFact IMP321, IS Patch, ISS, ISCMATRIX, JuvImmune, LipoVac, MF59, monophospholipid A, Montanide IMS 1312, Montanide ISA 206, Montanide ISA 50V, Montanide... ISA-51, OK-432, OM-174, OM-197-MP-EC, ONTAK, PepTel vector system, PLG microparticles, Remiquimod, SRL172, viral particles and other virus-like particles, YF-17D, VEGFtrap, R848, β-glucan, Pam3Cys, saponin-derived Aquila QS21 stimulator (Aquila Biotech, Worcester, Mass., USA), mycobacterial extracts and synthetic bacterial cell wall mimics, and other proprietary adjuvants such as Ribi's Detox.Quil or Superfos. Adjuvants (such as incomplete Freund's or GM-CSF) are also useful. Several dendritic cell-specific immune adjuvants (e.g., MF59) and their preparation have been described previously (Dupuis M, et al., Cell Immunol. 1998; 186(1):18-27; Allison AC; Dev Biol Stand. 1998; 92:3-11). Cytokines may also be used. Several cytokines have been directly linked to influence the migration of dendritic cells to lymphoid tissues (e.g., TNF-α), accelerate the maturation of dendritic cells into effective antigen-presenting cells (e.g., GM-CSF, IL-1, and IL-4) (US Patent No. 5,849,589, which is incorporated herein by reference in its entirety), and act as immune adjuvants (e.g., IL-12) (Gabrilovich DI et al., J Immunother Emphasis Tumor Immunol. 1996(6):414-418). HLA surface expression and intracellular protein processing into peptides to be presented on HLA can also be enhanced by interferon-γ (IFN-γ).See, for example, York IA, Goldberg AL, Mo XY, Rock KL. Proteolysis and class I majorhistocompatibility 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, which are incorporated herein by reference in their entirety.

[0325] HLA-peptide ABP

[0326] This article also provides ABPs that specifically bind to the HLA-peptide targets described herein.

[0327] HLA-peptide targets can be expressed on the surface of any suitable target cells, including tumor cells.

[0328] The ABP can specifically bind to human leukocyte antigen (HLA)-peptide targets, wherein the HLA-peptide targets include HLA-restricted peptides complexed with HLA class I molecules, wherein the HLA-restricted peptides are located in the peptide-binding groove of the α1 / α2 heterodimer portion of the HLA class I molecule.

[0329] In some respects, ABP does not bind to HLA class I in the absence of an HLA-restricted peptide. In some respects, ABP does not bind to an HLA-restricted peptide in the absence of human MHC class I. In some respects, ABP binds to tumor cells presenting human MHC class I complexed with an HLA-restricted peptide, optionally wherein the HLA-restricted peptide is a tumor antigen characterizing cancer.

[0330] ABPs can bind to each part of an HLA-peptide complex (i.e., HLA and a peptide representing each part of the complex). When bound together, they form new targets and protein surfaces that interact with and are bound by the ABP, unlike the surfaces presented by individual peptides or individual HLA subtypes. Generally, in the absence of each part of the HLA-peptide complex, there are no new targets and protein surfaces formed by HLA binding to peptides.

[0331] ABP can specifically bind to a complex containing HLA and an HLA-restricted peptide (HLA-peptide), for example, derived from a tumor. In some aspects, ABP does not bind to HLA in the absence of a tumor-derived HLA-restricted peptide. In other aspects, ABP does not bind to tumor-derived HLA-restricted peptides in the absence of HLA. In other aspects, ABP binds to a complex containing HLA and an HLA-restricted peptide when the HLA-restricted peptide is naturally present on cells (such as tumor cells).

[0332] In some implementations, the ABP provided herein regulates the binding of HLA-peptides to one or more ligands of the HLA-peptides.

[0333] ABP can specifically bind to any of the HLA-peptide targets disclosed in Table A, Table A1, or Table A2. In some embodiments, the HLA-restricted peptide is not derived from a gene selected from WT1 or MART1. In some embodiments, the ABP does not specifically bind to any of the target numbers 6364-6369, 6386-6389, 6500, 6521-6524, or 6578, and does not specifically bind to the HLA-peptide targets present in Table B or Table C.

[0334] In a more specific embodiment, ABP specifically binds to an HLA-peptide target selected from any of the following: HLA subtype A*02:01 conjugated with an HLA-restricted peptide containing the sequence LLASSILCA (SEQ ID NO:322); HLA subtype A*01:01 conjugated with an HLA-restricted peptide containing the sequence EVDPIGHLY (SEQ ID NO:354); HLA subtype B*44:02 conjugated with an HLA-restricted peptide containing the sequence GEMSSNSTAL (SEQ ID NO:357); HLA subtype A*02:01 conjugated with an HLA-restricted peptide containing the sequence GVYDGEEHSV (SEQ ID NO:356); HLA subtype *01:01 conjugated with an HLA-restricted peptide containing the sequence EVDPIGHVY (SEQ ID NO:2); and HLA subtype B*01:01 conjugated with an HLA-restricted peptide containing the sequence NTDNNLAVY (SEQ ID NO:322). HLA subtype HLA-A*01:01, which contains an HLA-restricted peptide complex with the sequence EVDPIGHVY (SEQ ID NO:2), HLA subtype B*35:01, which contains an HLA-restricted peptide complex with the sequence AIFPGAVPAA (SEQ ID NO:198), HLA subtype HLA-A*02:01, which contains an HLA-restricted peptide complex with the sequence ASSLPTTMNY (SEQ ID NO:260), and HLA subtype A*01:01.

[0335] In a more specific embodiment, ABP specifically binds to an HLA-peptide target selected from any of the following: HLA subtype A*02:01 composed of an HLA-restricted peptide consisting essentially of the sequence LLASSILCA (SEQ ID NO:322); HLA subtype A*01:01 composed of an HLA-restricted peptide consisting essentially of the sequence EVDPIGHLY (SEQ ID NO:354); HLA subtype B*44:02 composed of an HLA-restricted peptide consisting essentially of the sequence GEMSSNSTAL (SEQ ID NO:357); HLA subtype A*02:01 composed of an HLA-restricted peptide consisting essentially of the sequence GVYDGEEHSV (SEQ ID NO:356); HLA subtype *01:01 composed of an HLA-restricted peptide consisting essentially of the sequence EVDPIGHVY (SEQ ID NO:2); and HLA subtype B*44:02 composed of an HLA-restricted peptide consisting essentially of the sequence NTDNNLAVY (SEQ ID NO:322). HLA-peptide targets of any one of the following: HLA-A*01:01, which is composed of an HLA-restricted peptide complex consisting of the sequence EVDPIGHVY (SEQ ID NO:2); HLA-A*02:01, which is composed of an HLA-restricted peptide complex consisting of the sequence AIFPGAVPAA (SEQ ID NO:198); and HLA-A*01:01, which is composed of an HLA-restricted peptide complex consisting of the sequence ASSLPTTMNY (SEQ ID NO:260).

[0336] In a more specific embodiment, ABP specifically binds to an HLA-peptide target selected from any of the following: HLA subtype A*02:01 composed of an HLA-restricted peptide consisting of the sequence LLASSILCA (SEQ ID NO:322); HLA subtype A*01:01 composed of an HLA-restricted peptide consisting of the sequence EVDPIGHLY (SEQ ID NO:354); HLA subtype B*44:02 composed of an HLA-restricted peptide consisting of the sequence GEMSSNSTAL (SEQ ID NO:357); HLA subtype A*02:01 composed of an HLA-restricted peptide consisting of the sequence GVYDGEEHSV (SEQ ID NO:356); HLA subtype *01:01 composed of an HLA-restricted peptide consisting of the sequence EVDPIGHVY (SEQ ID NO:2); and HLA subtype B*01:01 composed of an HLA-restricted peptide consisting of the sequence NTDNNLAVY (SEQ ID NO:322). HLA subtypes HLA-A*01:01 (composed of HLA-restricted peptide complex with sequence NO:73), B*35:01 (composed of HLA-restricted peptide complex with sequence EVDPIGHVY (SEQ ID NO:2), A*02:01 (composed of HLA-restricted peptide complex with sequence AIFPGAVPAA (SEQ ID NO:198), and A*01:01 (composed of HLA-restricted peptide complex with sequence ASSLPTTMNY (SEQ ID NO:260)).

[0337] In some implementations, the ABP is a competing ABP with the illustrative ABP provided herein. In some aspects, the competing ABP and the illustrative ABP provided herein combine with the same epitope.

[0338] In some embodiments, the ABP described herein is referred to herein as a “variant.” In some embodiments, such a variant is derived from the sequence provided herein, for example, by affinity maturation, site-directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such a variant is not derived from the sequence provided herein, but can be de novo isolated, for example, according to the methods provided herein for obtaining an ABP. In some embodiments, the variant is derived from any sequence provided herein, wherein one or more conserved amino acid substitutions are performed. In some embodiments, the variant is derived from any sequence provided herein, wherein one or more non-conserved amino acid substitutions are performed. Conserved amino acid substitutions are described herein. Exemplary non-conserved amino acid substitutions include those described in the following literature: J Immunol. 2008 May 1; 180(9):6116-31, which is incorporated herein by reference in its entirety. In a preferred embodiment, the non-conserved amino acid substitution does not interfere with or inhibit the biological activity of the functional variant. In a more preferred embodiment, the non-conserved amino acid substitution enhances the biological activity of the functional variant, thereby enhancing the biological activity of the functional variant relative to the parental ABP.

[0339] ABP containing an antibody or its antigen-binding fragment

[0340] ABP may contain antibodies or their antigen-binding fragments.

[0341] In some implementations, the ABP provided herein comprises a light chain. In some aspects, the light chain is a κ light chain. In other aspects, the light chain is a λ light chain.

[0342] In some embodiments, the ABP provided herein comprises a heavy chain. In some aspects, the heavy chain is IgA. In some aspects, the heavy chain is IgD. In some aspects, the heavy chain is IgE. In some aspects, the heavy chain is IgG. In some aspects, the heavy chain is IgM. In some aspects, the heavy chain is IgG1. In some aspects, the heavy chain is IgG2. In some aspects, the heavy chain is IgG3. In some aspects, the heavy chain is IgG4. In some aspects, the heavy chain is IgA1. In some aspects, the heavy chain is IgA2.

[0343] In some embodiments, the ABP provided herein comprises an antibody fragment. In some embodiments, the ABP provided herein consists of an antibody fragment. In some embodiments, the ABP provided herein consists primarily of an antibody fragment. In some aspects, the ABP fragment is an Fv fragment. In some aspects, the ABP fragment is a Fab fragment. In some aspects, the ABP fragment is an F(ab')2 fragment. In some aspects, the ABP fragment is a Fab' fragment. In some aspects, the ABP fragment is an scFv (sFv) fragment. In some aspects, the ABP fragment is an scFv-Fc fragment. In some aspects, the ABP fragment is a fragment of a single-domain ABP.

[0344] In some implementations, the ABP fragments provided herein are derived from the illustrative ABPs provided herein. In other implementations, the ABP fragments provided herein are not derived from the illustrative ABPs provided herein, but can be extracted from scratch, for example, according to the methods provided herein for obtaining ABP fragments.

[0345] In some embodiments, the ABP fragments provided herein retain the ability to bind to HLA-peptide targets, such as by one or more assays or biological effects described herein. In some embodiments, the ABP fragments provided herein retain the ability to prevent HLA-peptides from interacting with one or more ligands, as described herein.

[0346] In some implementations, the ABP provided herein is a monoclonal ABP. In some implementations, the ABP provided herein is a polyclonal ABP.

[0347] In some embodiments, the ABP provided herein includes a chimeric ABP. In some embodiments, the ABP provided herein consists of a chimeric ABP. In some embodiments, the ABP provided herein consists primarily of a chimeric ABP. In some embodiments, the ABP provided herein includes a humanized ABP. In some embodiments, the ABP provided herein consists of a humanized ABP. In some embodiments, the ABP provided herein consists primarily of a humanized ABP. In some embodiments, the ABP provided herein includes a human ABP. In some embodiments, the ABP provided herein consists of a human ABP. In some embodiments, the ABP provided herein consists primarily of a human ABP.

[0348] In some embodiments, the ABP provided herein includes a replacement stent. In some embodiments, the ABP provided herein consists of a replacement stent. In some embodiments, the ABP provided herein consists essentially of a replacement stent. Any suitable replacement stent can be used. In some aspects, the replacement stent is selected from: Adnectin TM iMab EETI-II / AGRP, Kunitz domain, thioredoxin peptide aptamer, DARPin, Affilin, Tetranectin, Fynomer and Avimer.

[0349] This article also discloses isolated humanized, human or chimeric ABPs that compete with the ABPs disclosed herein for binding to HLA-peptides.

[0350] This article also discloses isolated humanized, human or chimeric ABPs that bind to the HLA-peptide epitopes of the ABPs described herein.

[0351] In some respects, ABP contains a human Fc region containing at least one modification that reduces binding to human Fc receptors.

[0352] It is known that when ABP is expressed in cells, it is post-translationally modified. Examples of post-translational modifications include: cleavage of lysine at the C-terminus of the heavy chain by carboxypeptidase; modification of the N-terminus of the heavy and light chains with glutamine or glutamate via pyroglutamylmethylation, glycosylation, oxidation, deamidation, and glycosylation, and such post-translational modifications are known to occur in various ABPs (see Journal of Pharmaceutical Sciences, 2008, Vol. 97, pp. 2426-2447, which is incorporated herein by reference in its entirety). In some embodiments, the ABP is a post-translational modified ABP or its antigen-binding fragment thereof. Examples of post-translational modified ABPs or their antigen-binding fragments thereof include ABPs or their antigen-binding fragments that have undergone pyroglutamylmethylation at the N-terminus of the variable region of the heavy chain and / or have lysine absent at the C-terminus of the heavy chain. It is known in the art that such post-translational modifications, resulting from N-terminal pyroglutamyl methylation and C-terminal lysine deletion, have no effect on the activity of ABP or fragments thereof (Analytical Biochemistry, 2006, Vol. 348, pp. 24-39, which are incorporated herein by reference in their entirety).

[0353] Single-specific and multi-specific HLA-peptide ABP

[0354] In some implementations, the ABP provided herein is a single-specific ABP.

[0355] In some implementations, the ABP provided herein is a multispecific ABP.

[0356] In some embodiments, the multispecific ABP provided herein binds to more than one antigen. In some embodiments, the multispecific ABP binds to two antigens. In some embodiments, the multispecific ABP binds to three antigens. In some embodiments, the multispecific ABP binds to four antigens. In some embodiments, the multispecific ABP binds to five antigens.

[0357] In some embodiments, the multispecific ABP provided herein binds to more than one epitope on the HLA-peptide antigen. In some embodiments, the multispecific ABP binds to two epitopes on the HLA-peptide antigen. In some embodiments, the multispecific ABP binds to three epitopes on the HLA-peptide antigen.

[0358] Many multispecific ABP constructs are known in the art, and the ABPs provided herein can be provided in any suitable form of multispecific construct.

[0359] In some embodiments, the multispecific ABP comprises an immunoglobulin containing at least two distinct heavy chain variable regions, each heavy chain variable region pairing with a common light chain variable region (i.e., the "common light chain ABP"). The common light chain variable region forms a distinct antigen-binding domain with each of the two distinct heavy chain variable regions. See Merchant et al., Nature Biotechnol., 1998, 16:677-681, which is incorporated herein by reference in its entirety.

[0360] In some embodiments, a multispecific ABP comprises an immunoglobulin, wherein the immunoglobulin comprises one or more fragments of the ABP linked to the N-terminus or C-terminus of the heavy or light chain of the immunoglobulin. See Coloma and Morrison, Nature Biotechnol., 1997, 15:159-163, which are incorporated herein by reference in their entirety. In some aspects, such an ABP comprises a tetravalent bispecific ABP.

[0361] In some embodiments, the multispecific ABP comprises a hybrid immunoglobulin containing at least two distinct heavy chain variable regions and at least two distinct light chain variable regions. See Milstein and Cuello Nature, 1983, 305:537-540; and Staerz and Bevan, Proc. Natl. Acad. Sci. USA, 1986, 83:1453-1457, each of which is incorporated herein by reference in its entirety.

[0362] In some embodiments, the multispecific ABP comprises a modified immunoglobulin chain to reduce the formation of non-multispecific byproducts. In some aspects, the ABP comprises one or more "mortar and pestle" modifications, as described in U.S. Patent No. 5,731,168, which is incorporated herein by reference in its entirety.

[0363] In some embodiments, the multispecific ABP comprises an immunoglobulin chain having one or more electrostatic modifications to facilitate the assembly of Fc heteromultimers. See WO 2009 / 089004, which is incorporated herein by reference in its entirety.

[0364] In some implementations, the multispecific ABP comprises a bispecific single-chain molecule. See Traunecker et al., EMBO J., 1991, 10: 3655-3659; and Gruber et al., J. Immunol., 1994, 152: 5368-5374, each incorporated herein by reference in its entirety.

[0365] In some embodiments, a multispecific ABP comprises a heavy chain variable domain and a light chain variable domain linked via a peptide linker, wherein the linker length is selected to facilitate the assembly of a multispecific ABP with the desired multispecificity. For example, when the heavy chain variable domain and the light chain variable domain are linked via a peptide linker having more than 12 amino acid residues, a monospecific scFv is typically formed. See U.S. Patents 4,946,778 and 5,132,405, both incorporated herein by reference in their entirety. In some embodiments, reducing the peptide linker length to less than 12 amino acid residues prevents the pairing of heavy chain and light chain variable domains on the same polypeptide chain, thereby allowing the heavy chain and light chain variable domains from one chain to pair with complementary domains on the other chain. Thus, the resulting ABP is multispecific, with the specificity at each binding site contributed by more than one polypeptide chain. Polypeptide chains comprising heavy chain and light chain variable domains linked by linkers of 3 to 12 amino acid residues primarily form dimers (called biantibodies). Linkers with 0 to 2 amino acid residues, i.e., trimers (called triantibodies) and tetramers (called tetraantibodies), are advantageous. However, apart from the length of the linker, the exact type of oligomerization appears to depend on the composition of the amino acid residues and the order of variable domains in each polypeptide chain (e.g., V...). H -Connector-V L With V L -Connector-V H Technicians can select the appropriate connector length based on the required multispecificity.

[0366] Fc region and variants

[0367] In some embodiments, the ABP provided herein comprises an Fc region. The Fc region may be wild-type or a variant thereof. In some embodiments, the ABP provided herein comprises an Fc region having one or more amino acid substitutions, insertions, or deletions compared to a naturally occurring Fc region. In some aspects, such substitutions, insertions, or deletions produce an ABP with altered stability, glycosylation, or other characteristics. In some aspects, such substitutions, insertions, or deletions produce a glycosylated ABP.

[0368] The “variant Fc region” or “engineered Fc region” comprises an amino acid sequence that differs from the native Fc region due to modification of at least one amino acid, preferably one or more amino acid substitutions. Preferably, the variant Fc region has at least one amino acid substitution, for example, about one to about ten amino acid substitutions, compared to the native Fc region or the Fc region of the parent polypeptide, and more preferably, about one to about five amino acid substitutions in the native Fc region or the Fc region of the parent polypeptide. The variant Fc region herein preferably has at least about 80% homology with the native Fc region and / or with the Fc region of the parent polypeptide, and most preferably, at least about 90% homology with it, more preferably, at least about 95% homology with it.

[0369] The term "ABP containing the Fc region" refers to an ABP that includes the Fc region. The C-terminal lysine (residue 447, according to the EU numbering system) of the Fc region can be removed, for example, during ABP purification or by recombinant engineering of the nucleic acid encoding the ABP. Therefore, an ABP with the Fc region can include ABPs with or without K447.

[0370] In some respects, the Fc region of the ABP provided herein is modified to produce an ABP with altered affinity for the Fc receptor, or to produce a more immune-inert ABP. In some embodiments, the ABP variants provided herein have some, but not all, effector functions. For example, such an ABP may be useful when the half-life of the ABP is important in vivo, but when certain effector functions (e.g., complement activation and ADCC) are unnecessary or harmful.

[0371] In some embodiments, the Fc region of the ABP provided herein is the human IgG4 Fc region, which contains one or more hinge-stabilizing mutations S228P and L235E. See Aalberse et al., Immunology, 2002, 105:9-19, which is incorporated herein by reference in its entirety. In some embodiments, the IgG4 Fc region contains one or more of the following mutations: E233P, F234V, and L235A. See Armour et al., Mol. Immunol., 2003, 40:585-593, which is incorporated herein by reference in its entirety. In some embodiments, the IgG4 Fc region contains a deletion at the G236 position.

[0372] In some embodiments, the Fc region of the ABP provided herein is the human IgG1 Fc region, which contains one or more mutations that reduce Fc receptor binding. In some aspects, said one or more mutations are in residues selected from S228 (e.g., S228A), L234 (e.g., L234A), L235 (e.g., L235A), D265 (e.g., D265A), and N297 (e.g., N297A). In some aspects, the ABP contains the PVA236 mutation. PVA236 refers to the substitution of PVA for the amino acid sequence ELLG (SEQ ID NO: 362) of IgG1 from amino acid positions 233 to 236 or for the EFLG (SEQ ID NO: 363) of IgG4. See U.S. Patent No. 9,150,641, which is incorporated herein by reference in its entirety.

[0373] In some implementations, the Fc region of the ABP provided herein is modified as described in the following references: Armour et al., Eur. J. Immunol., 1999, 29: 2613-2624; WO 1999 / 058572; and / or UK Patent Application No. 98099518, each of which is incorporated herein by reference in its entirety.

[0374] In some implementations, the Fc region of the ABP provided herein is the human IgG2 Fc region, which contains one or more mutations A330S and P331S.

[0375] In some embodiments, the Fc region of the ABP provided herein has amino acid substitutions at one or more of the following positions: 238, 265, 269, 270, 297, 327, and 329. See U.S. Patent No. 6,737,056, which is incorporated herein by reference in its entirety. Such Fc mutants comprise Fc mutants having substitutions at two or more of the amino acid positions 265, 269, 270, 297, and 327, including the so-called “DANA” Fc mutant, which has alanine substitutions at residues 265 and 297. See U.S. Patent No. 7,332,581, which is incorporated herein by reference in its entirety. In some embodiments, the ABP comprises alanine at amino acid position 265. In some embodiments, the ABP comprises alanine at amino acid position 297.

[0376] In some embodiments, the ABP provided herein comprises an Fc region having one or more amino acid substitutions that improve ADCC, such as substitutions at one or more positions of Fc region 298, 333, and 334. In some embodiments, the ABP provided herein comprises an Fc region having one or more amino acid substitutions at positions 239, 332, and 330, as described in the following reference: Lazar et al., Proc. Natl. Acad. Sci. USA, 2006, 103: 4005-4010, which is incorporated herein by reference in its entirety.

[0377] In some implementations, the ABP provided herein includes one or more modifications that improve or reduce C1q binding and / or CDC. See U.S. Patent No. 6,194,551; WO 99 / 51642; and Idusogie et al., J. Immunol., 2000, 164:4178-4184; all of which are incorporated herein by reference in their entirety.

[0378] In some embodiments, the ABP provided herein comprises one or more modifications to increase half-life. ABPs having increased half-life and improved binding to neonatal Fc receptors (FcRn) are described, for example, in Hinton et al., J. Immunol., 2006, 176:346-356; and U.S. Patent Publication No. 2005 / 0014934; both of which are incorporated herein by reference in their entirety. Such Fc variants include Fc variants having substituted Fc residues at one or more of the following Fc regions of IgG: 238, 250, 256, 265, 272, 286, 303, 305, 307, 311, 312, 314, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424, 428, and 434. In some embodiments, the ABP comprises one or more non-Fc modifications that extend half-life. Exemplary non-Fc modifications for extending half-life are described, for example, in US20170218078, which is hereby incorporated herein by reference in its entirety.

[0379] In some implementations, the ABP provided herein includes one or more Fc region variants, as described in the following documents: U.S. Patents 7,371,826, 5,648,260, and 5,624,821; Duncan and Winter, Nature, 1988, 322:738-740; and WO 94 / 29351; each of which is incorporated herein by reference in its entirety.

[0380] An antibody specific to B*35:01_EVDPIGHVY (SEQ ID NO:2) (HLA-peptide target "G5").

[0381] In some respects, this document provides an ABP comprising an antibody or antigen-binding fragment thereof that specifically binds to an HLA-peptide target, wherein the HLA class I molecule of the HLA-peptide target is HLA subtype B*35:01, and the HLA-restricted peptide of the HLA-peptide target comprises or is composed of or substantially comprises the sequence EVDPIGHVY (SEQ ID NO:2) (“G5”).

[0382] CDR

[0383] ABP specific to B*35:01_EVDPIGHVY (SEQ ID NO:2) may contain one or more antibody complementarity-determining region (CDR) sequences, for example, it may contain three heavy chain CDRs (CDR-H1, CDR-H2, CDR-H3) and three light chain CDRs (CDR-L1, CDR-L2, CDR-L3).

[0384] ABP specific to B*35:01_EVDPIGHVY (SEQ ID NO:2) may contain the CDR-H3 sequence. The CDR-H3 sequence can be selected from the group consisting of CARDGVRYYGMDVW (SEQ ID NO: 3), CARGVRGYDRSAGYW (SEQ ID NO: 4), CASHDYGDYGEYFQHW (SEQ ID NO: 5), CARVSWYCSSTSCGVNWFDPW (SEQ ID NO: 6), CAKVNWNDGPYFDYW (SEQ ID NO:7), CAPTNSGYYGPYYYYGMDVW (SEQ ID NO:8), CARDVMDVW (SEQ ID NO:9), CAREGYGMDVW (SEQ ID NO:10), CARDNGVGVDYW (SEQ ID NO:11), CARGIADSGSYYGNGRDYYYGMDVW (SEQ ID NO:12), CARGDYYFDYW (SEQ ID NO:12) NO:13), CARDGTRYYGMDVW (SEQ ID NO:14), CARDVVANFDYW (SEQ ID NO:15), CARGHSSGWYYYYGMDVW (SEQ ID NO:16), CAKDLGSYGGYYW (SEQ ID NO:17), CARSWFGGFNYHYYGMDVW (SEQ ID NO:18), CARELPIGYGMDVW (SEQ ID NO:19) and CARGGSYYYYGMDVW (SEQ ID NO:20).

[0385] ABP specific to B*35:01_EVDPIGHVY (SEQ ID NO:2) may contain a CDR-L3 sequence. The CDR-L3 sequence may be selected from CMQGLQTPITF (SEQ ID NO:21), CMQALQTPPTF (SEQ ID NO:22), CQQAISFPLTF (SEQ ID NO:23), CQQANSFPLTF (SEQ ID NO:24), CQQANSFPLTF (SEQ ID NO:24), CQQSYSIPLTF (SEQ ID NO:24) NO:25), CQQTYMMPYTF (SEQ ID NO:26), CQQSYITPWTF (SEQ ID NO:27), CQQSYITPYTF (SEQ ID NO:28), CQQYYTTPYTF (SEQ ID NO:29), CQQSYSTPLTF (SEQ ID NO:30), CMQALQTPLTF (SEQ ID NO:31), CQQYGSWPRTF (SEQ ID NO:32), CQQSYSTPVTF (SEQ ID NO:33), CMQALQTPYTF (SEQ ID NO:34), CQQANSFPFTF (SEQ ID NO:35), CMQALQTPLTF (SEQ ID NO:31) and CQQSYSTPLTF (SEQ ID NO:30).

[0386] ABP specific to B*35:01_EVDPIGHVY (SEQ ID NO:2) may contain a specific heavy chain CDR3 (CDR-H3) sequence and a specific light chain CDR3 (CDR-L3) sequence. In some implementations, the ABP comprises CDR-H3 and CDR-L3 from the scFv, which is named G5_P7_E7, G5_P7_B3, G5_P7_A5, G5_P7_F6, G5-P1B12, G5-P1C12, G5-P1-E05, G5-P3G01, G5-P3G08, G5-P4B02, G5-P4E04, G5R4-P1D06, G5R4-P1H11, G5R4-P2B10, G5R4-P2H8, G5R4-P3G05, G5R4-P4A07, or G5R4-P4B01. Table 5 shows the CDR sequences of the identified scFv that specifically binds to B*35:01_EVDPIGHVY (SEQ ID NO:2). For clarity, each identified scFv is named with a clone name, and each line contains the CDR sequence for that specific clone name. For example, the clone identified by clone name G5_P7_E7... It contains the heavy chain CDR3 sequence CARDGVRYYGMDVW (SEQ ID NO:3) and the light chain CDR3 sequence CMQGLQTPITF (SEQ ID NO:21).

[0387] An ABP specific to B*35:01_EVDPIGHVY (SEQ ID NO:2) may contain all six CDRs from scFv, which are named G5_P7_E7, G5_P7_B3, G5_P7_A5, G5_P7_F6, G5-P1B12, G5-P1C12, G5-P1-E05, G5-P3G01, G5-P3G08, G5-P4B02, G5-P4E04, G5R4-P1D06, G5R4-P1H11, G5R4-P2B10, G5R4-P2H8, G5R4-P3G05, G5R4-P4A07, or G5R4-P4B01.

[0388] VH

[0389] An ABP specific to B*35:01_EVDPIGHVY (SEQ ID NO:2) may contain a VH sequence. The VH sequence may be selected from QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYDINWVRQAPGQGLEWMGIINPRSGSTKYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGVRYYGMDVWGQGTTVTVSS (SEQ ID NO:36), QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSHDINWVRQAPGQGLEWMGWMNPNSGDTGYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGVRGYDRSAGYWGQGTLVIVSS (SEQ ID NO:37), EVQLLESGGGLVKPGGSLRLSCAASGFSFSSYWMSWVRQAPGKGLEWISYISGDSGYTNYADSVKGRFTISRDDSKNTLYLQMNSLKTEDTAVYYCASHDYGDYGEYFQHWGQGTLVTVSS (SEQ ID NO:38), EVQLLQSGGGLVQPGGSLRLSCAASGFTFSNSDMNWVRQAPGKGLEWVAYISSGSSTIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVSWYCSSTSCGVNWFDPWGQGTLVTVSS (SEQ ID NO:39), EVQLLESGGGLVQPGGSLRLSCAASGFTFSNSDMNWVRQAPGKGLEWVASISSSGGYINYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKVNWNDGPYFDYWGQGTLVTVSS (SEQ ID NO:40), QVQLVQSGAEVKKPGSSVKVSCKASGGTFSNFGVSWLRQAPGQGLEWMGGIIPILGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCATPTNSGYYGPYYYYGMDVWGQGTTVTVSS (SEQ ID NO:41), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDVMDVWGQGTTVTVSS (SEQ IDNO:42)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSGYLVSWVRQAPGQGLEWMGWINPNSGGTNTAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAREGYGMDVWGQGTTVTVSS(SEQ ID NO:43)、QVQLVQSGAEVKKPGASVKVSCKASGYIFRNYPMHWVRQAPGQGLEWMGWINPDSGGTKYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDNGVGVDYWGQGTLVTVSS(SEQ ID NO:44)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWMNPNIGNTGYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGIADSGSYYGNGRDYYYGMDVWGQGTTVTVSS(SEQ ID NO:45)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSSYGISWVRQAPGQGLEWMGWINPNSGVTKYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGDYYFDYWGQGTLVTVSS(SEQ ID NO:46)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYDINWVRQAPGQGLEWMGWINPNSGDTKYSQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGTRYYGMDVWGQGTTVTVSS(SEQ ID NO:47)、EVQLLESGGGLVKPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVSYISSSSSYTNYADSVKGRFTISRDDSKNTLYLQMNSLKTEDTAVYYCARDVVANFDYWGQGTLVTVSS(SEQ ID NO:48)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGWMNPDSGSTGYAQRFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGHSSGWYYYYGMDVWGQGTTVTVSS(SEQ IDNO:49), EVQLLESGGGLVQPGGSLRLSCAASGFTFTSYSMHWVRQAPGKGLEWVSSITSFTNTMYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDLGSYGGYYWGQGTLVTVSS (SEQ ID NO:50), QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARSWFGGFNYHYYGMDVWGQGTTVTVSS (SEQ ID NO:51), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGWMNPNSGNTGYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARELPIGYGMDVWGQGTTVTVSS (SEQ ID NO:52) and QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIVGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGGSYYYYGMDVWGQGTTVTVSS (SEQ ID NO:53).

[0390] VL

[0391] An ABP specific to B*35:01_EVDPIGHVY (SEQ ID NO:2) may comprise a VL sequence. The VL sequence may be selected from DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSYRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGLQTPITFGQGTRLEIK (SEQ ID NO:54), DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSSRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTPPTFGPGTKVDIK (SEQ ID NO:55), DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQAISFPLTFGQSTKVEIK (SEQ ID NO:56), DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYSASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPLTFGGGTKVEIK (SEQ ID NO:57), DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPLTFGGGTKVEIK (SEQ ID NO:58), DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSIPLTFGGGTKVEIK (SEQ ID NO:59), DIQMTQSPSSLSASVGDRVTITCRASQGISNYLNWYQQKPGKAPKLLIYYASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTYMMPYTFGQGTKVEIK (SEQ IDNO:60), DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYGASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYITPWTFGQGTKVEIK(SEQ ID NO:61), DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYITPYTFGQGTKLEIK(SEQ ID NO:62), DIVMTQSPDSLAVSLGERATINCKTSQSVLYRPNNENYLAWYQQKPGQPPKLLIYQASIREPGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYTTPYTFGQGTKLEIK(SEQ ID NO:63), DIQMTQSPSSLSASVGDRVTITCRASQSISRFLNWYQQKPGKAPKLLIYGASRPQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGQGTKVEIK(SEQ ID NO:64) NO:64)、DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSHRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTPLTFGGGTKVEIK(SEQ ID NO:65)、EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYAASARASGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYGSWPRTFGQGTKVEIK(SEQ ID NO:66)、DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYGASRLQSGVPSRFGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPVTFGQGTKVEIK(SEQ ID NO:67)、DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTPYTFGQGTKVEIK(SEQ IDNO: 68), DIQMTQSPSSLSASVGDRVTITCQASEDISNHLNWYQQKPGKAPKLLIYDALSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPFTFGPGTKVDIK (SEQ ID NO: 69), DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTPLTFGQGTKVEIK (SEQ ID NO: 70) and DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIK (SEQ ID NO: 71).

[0392] VH-VL combination

[0393] ABP specific to B*35:01_EVDPIGHVY (SEQ ID NO:2) may contain specific VH sequences and specific VL sequences. In some implementations, the ABP specific to B*35:01_EVDPIGHVY (SEQ ID NO:2) comprises a VH sequence and a VL sequence from an scFv named G5_P7_E7, G5_P7_B3, G5_P7_A5, G5_P7_F6, G5-P1B12, G5-P1C12, G5-P1-E05, G5-P3G01, G5-P3G08, G5-P4B02, G5-P4E04, G5R4-P1D06, G5R4-P1H11, G5R4-P2B10, G5R4-P2H8, G5R4-P3G05, G5R4-P4A07, or G5R4-P4B01. Table 4 shows the VH and VL sequences of identified scFvs that specifically bind to B*35:01_EVDPIGHVY (SEQ ID NO:2). For clarity, each identified scFv hit is named with a clone name, and each row contains the VH and VL sequences for that specific clone name. For example, the scFv identified by the clone name G5_P7_E7 contains the VH sequence QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYDINWVRQAPGQGLEWMGIINPRSGSTKYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGVRYYGMDVWGQGTTVTVSS (SEQ ID NO:36) and the VL sequence DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSYRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGLQTPITFGQGTRLEIK (SEQ ID NO:54).

[0394] Antibodies specific to A*02:01_AIFPGAVPAA (SEQ ID NO:198) (HLA-peptide target "G8")

[0395] In some respects, this document provides an ABP comprising an antibody or antigen-binding fragment thereof that specifically binds to an HLA-peptide target, wherein the HLA class I molecule of the HLA-peptide target is HLA subtype A*02:01, and the HLA-restricted peptide of the HLA-peptide target comprises, or is substantially composed of, the sequence AIFPGAVPAA (SEQ ID NO:198) (“G8”).

[0396] CDR

[0397] ABP specific to A*02:01_AIFPGAVPAA (SEQ ID NO:198) may contain one or more antibody complementarity-determining region (CDR) sequences, such as three heavy chain CDRs (CDR-H1, CDR-H2, CDR-H3) and three light chain CDRs (CDR-L1, CDR-L2, CDR-L3).

[0398] ABP specific to A*02:01_AIFPGAVPAA (SEQ ID NO:198) may contain the CDR-H3 sequence. The CDR-H3 sequence may be selected from the group consisting of CARDDYGDYVAYFQHW (SEQ ID NO: 201), CARDLSYYYGMDVW (SEQ ID NO: 202), CARVYDFWSVLSGFDIW (SEQ ID NO: 203), CARVEQGYDIYYYYYMDVW (SEQ ID NO: 204), CARSYDYGDYLNFDYW (SEQ ID NO: 204) NO:205),CARASGSGYYYYYGMDVW(SEQ ID NO:206),CAASTWIQPFDYW(SEQ ID NO:207),CASNGNYYGSGSYYNYW(SEQ ID NO:208),CARAVYYDFWSGPFDYW(SEQ ID NO:209),CAKGGIYYGSGSYPSW(SEQ ID NO:210), CARGLYYMDVW (SEQ ID NO:211), CARGLYGDYFLYYGMDVW (SEQ ID NO:212), CARGLLGFGEFLTYGMDVW (SEQ ID NO:213), CARDRDSSWTYYYYGMDVW (SEQ ID NO:214), CARGLYGDYFLYYGMDVW (SEQ ID NO:212), CARGDYYDSSGYYFPVYFDYW (SEQ ID NO:215), and CAKDPFWSGHYYYYGMDVW (SEQ ID NO:212) ID NO:216).

[0399] ABP specific to A*02:01_AIFPGAVPAA (SEQ ID NO:198) may contain a CDR-L3 sequence. The CDR-L3 sequence can be selected from CQQNYNSVTF (SEQ ID NO:217), CQQSYNTPWTF (SEQ ID NO:218), CGQSYSTPPTF (SEQ ID NO:219), CQQSYSAPYTF (SEQ ID NO:111), CQQSYSIPPTF (SEQ ID NO:220), CQQSYSAPYTF (SEQ ID NO:220) NO:111), CQQHNSYPPTF (SEQ ID NO:221), CQQYSTYPITI (SEQ ID NO:222), CQQANSFPWTF (SEQ ID NO:223), CQQSHSTPQTF (SEQ ID NO:224), CQQSYSTPLTF (SEQ ID NO:30), CQQSYSTPLTF (SEQ ID NO:30), CQQTYSTPWTF (SEQ ID NO:225), CQQYGSSPYTF (SEQ ID NO:226), CQQSHSTPLTF (SEQ ID NO:227), CQQANGFPLTF (SEQ ID NO:228) and CQQSYSTPLTF (SEQ ID NO:30).

[0400] ABPs specific to A*02:01_AIFPGAVPAA (SEQ ID NO:198) may contain a specific heavy chain CDR3 (CDR-H3) sequence and a specific light chain CDR3 (CDR-L3) sequence. In some implementations, the ABP comprises CDR-H3 and CDR-L3 from scFv, which are named G8-P1A03, G8-P1A04, G8-P1A06, G8-P1B03, G8-P1C11, G8-P1D02, G8-P1H08, G8-P2B05, G8-P2E06, R3G8-P2C10, R3G8-P2E04, R3G8-P4F05, R3G8-P5C03, R3G8-P5F02, R3G8-P5G08, G8-P1C01, or G8-P2C11. Table 7 shows the CDR sequences of identified scFvs that specifically bind to A*02:01_AIFPGAVPAA (SEQ ID NO:198). For clarity, each identified scFv match is named with a clone name, and each line contains the CDR sequence for that specific clone name. For example, the scFv identified by clone name G8-P1A03 contains the heavy chain CDR3 sequence CARDDYGDYVAYFQHW (SEQ ID NO:201) and the light chain CDR3 sequence CQQNYNSVTF (SEQ ID NO:217).

[0401] ABP specific to A*02:01_AIFPGAVPAA (SEQ ID NO:198) may contain all 6 CDRs from scFv, which are named G8-P1A03, G8-P1A04, G8-P1A06, G8-P1B03, G8-P1C11, G8-P1D02, G8-P1H08, G8-P2B05, G8-P2E06, R3G8-P2C10, R3G8-P2E04, R3G8-P4F05, R3G8-P5C03, R3G8-P5F02, R3G8-P5G08, G8-P1C01, or G8-P2C11.

[0402] VH

[0403] An ABP specific for A*02:01_AIFPGAVPAA (SEQ ID NO:198) may comprise a VH sequence. The VH sequence may be selected from QVQLVQSGAEVKKPGASVKVSCKASGGTFSRSAITWVRQAPGQGLEWMGWINPNSGATNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDDYGDYVAYFQHWGQGTLVTVSS (SEQ ID NO:229), QVQLVQSGAEVKKPGASVKVSCKASGYPFIGQYLHWVRQAPGQGLEWMGIINPSGDSATYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDLSYYYGMDVWGQGTTVTVSS (SEQ ID NO:230), QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYMHWVRQAPGQGLEWMGWMNPIGGGTGYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARVYDFWSVLSGFDIWGQGTLVTVSS (SEQ ID NO:231), EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVSGINWNGGSTGYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVEQGYDIYYYYYMDVWGKGTTVTVSS (SEQ ID NO:232), QVQLVQSGAEVKKPGASVKVSCKASGGTLSSYPINWVRQAPGQGLEWMGWISTYSGHADYAQKLQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARSYDYGDYLNFDYWGQGTLVTVSS (SEQID NO:199), EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYWMSWVRQAPGKGLEWVSSISGRGDNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARASGSGYYYYYGMDVWGQGTTVTVSS (SEQ IDNO:233)、QVQLVQSGAEVKKPGASVKVSCKASGYTFGNYFMHWVRQAPGQGLEWMGMVNPSGGSETFAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAASTWIQPFDYWGQGTLVTVSS(SEQ ID NO:234)、EVQLLESGGGLVQPGGSLRLSCAASGFDFSIYSMNWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASNGNYYGSGSYYNYWGQGTLVTVSS(SEQ ID NO:235)、QVQLVQSGAEVKKPGASVKVSCKASGYTLTTYYMHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARAVYYDFWSGPFDYWGQGTLVTVSS(SEQ ID NO:236)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGWINPYSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKGGIYYGSGSYPSWGQGTLVTVSS(SEQID NO:237)、QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYGVSWVRQAPGQGLEWMGWISPYSGNTDYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGLYYMDVWGKGTTVTVSS(SEQ ID NO:238)、QVQLVQSGAEVKKPGASVKVSCKASGYTFSNMYLHWVRQAPGQGLEWMGWINPNTGDTNYAQTFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGLYGDYFLYYGMDVWGQGTKVTVSS(SEQ ID NO:239)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGWMNPNSGNTGYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGLLGFGEFLTYGMDVWGQGTLVTVSS(SEQ IDNO: 240), QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYIHWVRQAPGQGLEWMGVINPSGGSTTYAQKLQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDRDSSWTYYYYGMDVWGQGTTVTVSS (SEQ ID NO: 241), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSNYMHWVRQAPGQGLEWMGWMNPNSGNTGYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGLYGDYFLYYGMDVWGQGTTVTVSS (SEQ ID NO: 242), QVQLVQSGAEVKKPGASVKVSCKASGGTFSSHAISWVRQAPGQGLEWMGVIIPSGGTSYTQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGDYYDSSGYYFPVYFDYWGQGTLVTVSS (SEQ ID NO: 243) and QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYAMNWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDPFWSGHYYYYGMDVWGQGTTVTVSS (SEQ ID NO: 244).

[0404] VL

[0405] An ABP specific to A*02:01_AIFPGAVPAA (SEQ ID NO:198) may contain a VL sequence. The VL sequence may be selected from DIQMTQSPSSLSASVGDRVTITCRASQSITSYLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQNYNSVTFGQGTKLEIK (SEQ ID NO:245), DIQMTQSPSSLSASVGDRVTITCWASQGISSYLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYNTPWTFGPGTKVDIK (SEQ ID NO:246), DIQMTQSPSSLSASVGDRVTITCRASQAISNSLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCGQSYSTPPTFGQGTKLEIK (SEQ ID NO:247), DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYKASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSAPYTFGPGTKVDIK (SEQ ID NO:248), DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSIPPTFGGGTKVDIK (SEQ ID NO:200), DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSAPYTFGGGTKVEIK (SEQ ID NO:249), DIQMTQSPSSLSASVGDRVTITCRASQGINSYLAWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHNSYPPTFGQGTKLEIK (SEQ IDNO:250)、DIQMTQSPSSLSASVGDRVTITCRASQSISRWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTYPITIGQGTKVEIK(SEQ ID NO:251)、DIQMTQSPSSLSASVGDRVTITCRASQGISNSLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPWTFGQGTKLEIK(SEQ ID NO:252)、DIQMTQSPSSLSASVGDRVTITCRASQDVSTWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSHSTPQTFGQGTKVEIK(SEQ ID NO:253)、DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKLEIK(SEQ ID NO:254)、DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIK(SEQ ID NO:255)、DIQMTQSPSSLSASVGDRVTITCRASQGISNWLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTYSTPWTFGQGTKLEIK(SEQID NO:256)、EIVMTQSPATLSVSPGERATLSCRASQSVGNSLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYGSSPYTFGQGTKVEIK(SEQ ID NO:257)、DIQMTQSPSSLSASVGDRVTITCRASQSISGYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSHSTPLTFGQGTKVEIK(SEQ IDNO: 258), DIQMTQSPSSLSASVGDRVTITCRASQNIYTYLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANGFPLTFGGGTKVEIK (SEQ ID NO: 259) and DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIK (SEQ ID NO: 71).

[0406] VH-VL combination

[0407] ABP specific to A*02:01_AIFPGAVPAA (SEQ ID NO:198) may contain a specific VH sequence and a specific VL sequence. In some implementations, the ABP specific to A*02:01_AIFPGAVPAA (SEQ ID NO:198) may comprise a VH sequence and a VL sequence from an scFv named G8-P1A03, G8-P1A04, G8-P1A06, G8-P1B03, G8-P1C11, G8-P1D02, G8-P1H08, G8-P2B05, G8-P2E06, R3G8-P2C10, R3G8-P2E04, R3G8-P4F05, R3G8-P5C03, R3G8-P5F02, R3G8-P5G08, G8-P1C01, or G8-P2C11. Table 6 shows the VH and VL sequences of identified scFvs that specifically bind to A*02:01_AIFPGAVPAA (SEQ ID NO:198). For clarity, each identified scFv hit is named with a clone name, and each row contains the VH and VL sequences for that specific clone name. For example, the scFv identified by clone name G8-P1A03 contains the VH sequence QVQLVQSGAEVKKPGASVKVSCKASGGTFSRSAITWVRQAPGQGLEWMGWINPNSGATNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDDYGDYVAYFQHWGQGTLVTVSS (SEQ ID NO:229) and the VL sequence DIQMTQSPSSLSASVGDRVTITCRASQSITSYLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQNYNSVTFGQGTKLEIK (SEQ ID NO:245).

[0408] Antibodies specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) (HLA-peptide target "G10")

[0409] In some respects, this document provides an ABP comprising an antibody or an antigen-binding fragment thereof that specifically binds to an HLA-peptide target, wherein the HLA class I molecule of the HLA-peptide target is HLA subtype A*01:01, and the HLA-restricted peptide of the HLA-peptide target comprises the sequence ASSLPTTMNY (SEQ ID NO:260) (“G10”), constitutes therewith, or is substantially composed of therewith.

[0410] CDR

[0411] ABP specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain one or more antibody complementarity-determining region (CDR) sequences, such as three heavy chain CDRs (CDR-H1, CDR-H2, CDR-H3) and three light chain CDRs (CDR-L1, CDR-L2, CDR-L3).

[0412] ABPs specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain the CDR-H3 sequence. The CDR-H3 sequence can be selected from CARDQDTIFGVVITWFDPW (SEQ ID NO:261), CARDKVYGDGFDPW (SEQ ID NO:262), CAREDDSMDVW (SEQ ID NO:263), CARDSSGLDPW (SEQ ID NO:264), CARGVGNLDYW (SEQID NO:265), CARDAHQYYDFWSGYYSGTYYYGMDVW (SEQ ID NO:266), CAREQWPSYWYFDLW (SEQ ID NO:267), CARDRGYSYGYFDYW (SEQ ID NO:268), CARGSGDPNYYYYYGLDVW (SEQ ID NO:269), CARDTGDHFDYW (SEQ ID NO:269) NO:270), CARAENGMDVW (SEQ ID NO:271), CARDPGGYMDVW (SEQ ID NO:272), CARDGDAFDIW (SEQ ID NO:273), CARDMGDAFDIW (SEQ ID NO:274), CAREEDGMDVW (SEQ ID NO:275), CARDTGDHFDYW (SEQ ID NO:270), CARGEYSSGFFFVGWFDLW (SEQ ID NO:276) and CARETGDDAFDIW (SEQ ID NO:277).

[0413] ABPs specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain a CDR-L3 sequence. The CDR-L3 sequence may be selected from CQQYFTTPYTF (SEQ ID NO:278), CQQAEAFPYTF (SEQ ID NO:279), CQQSYSTPITF (SEQ ID NO:280), CQQSYIIPYTF (SEQ ID NO:281), CHQTYSTPLTF (SEQ ID NO:282), CQQAYSFPWTF (SEQ ID NO:283), CQQGYSTPLTF (SEQ ID NO:284), CQQANSFPRTF (SEQ ID NO:285), CQQANSLPYTF (SEQ ID NO:286), CQQSYSTPFTF (SEQ ID NO:109), CQQSYSTPFTF (SEQ ID NO:109), CQQSYGVPTF (SEQ ID NO:287), CQQSYSTPLTF (SEQ ID NO:30), CQQSYSTPLTF (SEQ ID NO:30), CQQYYSYPWTF (SEQ ID NO:288), CQQSYSTPFTF (SEQ ID NO:109), CMQTLKTPLSF (SEQ ID NO:289) and CQQSYSTPLTF (SEQ ID NO:30).

[0414] An ABP specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain a specific heavy chain CDR3 (CDR-H3) sequence and a specific light chain CDR3 (CDR-L3) sequence. In some embodiments, the ABP contains CDR-H3 and CDR-L3 from scFvs named R3G10-P1A07, R3G10-P1B07, R3G10-P1E12, R3G10-P1F06, R3G10-P1H01, R3G10-P1H08, R3G10-P2C04, R3G10-P2G11, R3G10-P3E04, R3G10-P4A02, R3G10-P4C05, R3G10-P4D04, R3G10-P4D10, R3G10-P4E07, R3G10-P4E12, R3G10-P4G06, R3G10-P5A08, or R3G10-P5C08. Table 9 shows the CDR sequences of identified scFvs that specifically bind to A*01:01_ASSLPTTMNY (SEQ ID NO:260). For clarity, each identified scFv hit is named with a clone name, and each row contains the CDR sequence for that specific clone name. For example, the scFv identified by clone name R3G10-P1A07 contains the heavy chain CDR3 sequence CARDQDTIFGVVITWFDPW (SEQ ID NO:261) and the light chain CDR3 sequence CQQYFTTPYTF (SEQ ID NO:278).

[0415] For A*01:01_ASSLPTTMNY (SEQ ID NO:260) A specific ABP may contain all 6 CDRs from scFv, which are named R3G10-P1A07, R3G10-P1B07, R3G10-P1E12, R3G10-P1F06, R3G10-P1H01, R3G10-P1H08, R3G10-P2C04, R3G10-P2G11, R3G10-P3E04, R3G10-P4A02, R3G10-P4C05, R3G10-P4D04, R3G10-P4D10, R3G10-P4E07, R3G10-P4E12, R3G10-P4G06, R3G10-P5A08, or R3G10-P5C08.

[0416] VH

[0417] The ABP specific for A*01:01_ASSLPTTMNY (SEQ ID NO:260) may comprise a VH sequence. The VH sequence may be selected from EVQLLESGGGLVKPGGSLRLSCAASGFTFSSYWMSWVRQAPGKGLEWVSGISARSGRTYYADSVKGRFTISRDDSKNTLYLQMNSLKTEDTAVYYCARDQDTIFGVVITWFDPWGQGTLVTVSS (SEQ ID NO:290), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIIHPGGGTTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDKVYGDGFDPWGQGTLVTVSS (SEQ ID NO:291), QVQLVQSGAEVKKPGASVKVSCKASGYIFTGYYMHWVRQAPGQGLEWMGMIGPSDGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAREDDSMDVWGKGTTVTVSS (SEQ ID NO:292), QVQLVQSGAEVKKPGASVKVSCKASGYTFIGYYMHWVRQAPGQGLEWMGMIGPSDGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDSSGLDPWGQGTLVTVSS (SEQ ID NO:293), QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGMIGPSDGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGVGNLDYWGQGTLVTVSS (SEQ ID NO:294), QVQLVQSGAEVKKPGASVKVSCKASGVTFSTSAISWVRQAPGQGLEWMGWISPYNGNTDYAQMLQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDAHQYYDFWSGYYSGTYYYGMDVWGQGTTVTVSS (SEQ IDNO:295)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSNSIINWVRQAPGQGLEWMGWMNPNSGNTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAREQWPSYWYFDLWGRGTLVTVSS(SEQ ID NO:296)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSTHDINWVRQAPGQGLEWMGVINPSGGSAIYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDRGYSYGYFDYWGQGTLVTVSS(SEQ ID NO:297)、QVQLVQSGAEVKKPGASVKVSCKASGNTFIGYYVHWVRQAPGQGLEWVGIINPNGGSISYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSGDPNYYYYYGLDVWGQGTTVTVSS(SEQ ID NO:298)、QVQLVQSGAEVKKPGASVKVSCKASGYTLSYYYMHWVRQAPGQGLEWMGMIGPSDGSTSYAQRFQGRVTMTRDTSTGTVYMELSSLRSEDTAVYYCARDTGDHFDYWGQGTLVTVSS(SEQ ID NO:299)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGIIGPSDGSTTYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARAENGMDVWGQGTTVTVSS(SEQ ID NO:300)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYVHWVRQAPGQGLEWMGIIAPSDGSTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDPGGYMDVWGKGTTVTVSS(SEQ ID NO:301)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYLHWVRQAPGQGLEWMGMIGPSDGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGDAFDIWGQGTMVTVSS(SEQ IDNO: 302), QVQLVQSGAEVKKPGSSVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGRISPSDGSTTYAPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDMGDAFDIWGQGTTVTVSS (SEQ ID NO: 303), QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGMIGPSDGSTSYAQRFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAREEDGMDVWGQGTTVTVSS (SEQ ID NO: 304), QVQLVQSGAEVKKPGASVKVSCKASGYTLSYYYMHWVRQAPGQGLEWMGMIGPSDGSTSYAQRFQGRVTMTRDTSTGTVYMELSSLRSEDTAVYYCARDTGDHFDYWGQGTLVTVSS (SEQ ID NO: 299), QVQLVQSGAEVKKPGSSVKVSCKASGGTFNNFAISWVRQAPGQGLEWMGGIIPIFDATNYAQKFQGRVTFTADESTSTAYMELSSLRSEDTAVYYCARGEYSSGFFFVGWFDLWGRGTQVTVSS (SEQ ID NO: 305) and QVQLVQSGAEVKKPGASVKVSCKASGYNFTGYYMHWVRQAPGQGLEWMGIIAPSDGSTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARETGDDAFDIWGQGTMVTVSS (SEQ ID NO: 306).

[0418] VL

[0419] The ABP specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may comprise a VL sequence. The VL sequence may be selected from DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYFTTPYTFGQGTKLEIK (SEQ ID NO:307), DIQMTQSPSSLSASVGDRVTITCRASQSISRWLAWYQQKPGKAPKLLIFDASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQAEAFPYTFGQGTKVEIK (SEQ ID NO:308), DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPITFGQGTRLEIK (SEQ ID NO:309), DIQMTQSPSSLSASVGDRVTITCRASQSISNYLNWYQQKPGKAPKLLIYKASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYIIPYTFGQGTKLEIK (SEQ ID NO:310), DIQMTQSPSSLSASVGDRVTITCRASQSISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQTYSTPLTFGQGTKVEIK (SEQ ID NO:311), DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYSASNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQAYSFPWTFGQGTKVEIK (SEQ ID NO:312), DIQMTQSPSSLSASVGDRVTITCRASQNISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYSTPLTFGQGTRLEIK (SEQ IDNO:313)、DIQMTQSPSSLSASVGDRVTITCRASQDISRYLAWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPRTFGQGTKVEIK(SEQ ID NO:314)、DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSLPYTFGQGTKVEIK(SEQ ID NO:315)、DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASTLQNGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKVDIK(SEQ ID NO:316)、DIQMTQSPSSLSASVGDRVTITCRASQRISSYLNWYQQKPGKAPKLLIYSASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKVDIK(SEQ ID NO:317)、DIQMTQSPSSLSASVGDRVTITCRASQSISSYLAWYQQKPGKAPKLLIYDASKLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYGVPTFGQGTKLEIK(SEQ ID NO:318)、DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIK(SEQID NO:319)、DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIK(SEQ ID NO:71)、DIQMTQSPSSLSASVGDRVTITCRASQGISTYLAWYQQKPGKAPKLLIYDASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYSYPWTFGQGTRLEIK(SEQ IDNO: 320), DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASTLQNGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKVDIK (SEQ ID NO: 316), DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQTLKTPLSFGGGTKVEIK (SEQ ID NO: 321) and DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIK (SEQ ID NO: 71).

[0420] VH-VL combination

[0421] ABP specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain a specific VH sequence and a specific VL sequence. In some embodiments, an ABP specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) contains a VH sequence and a VL sequence from an scFv named R3G10-P1A07, R3G10-P1B07, R3G10-P1E12, R3G10-P1F06, R3G10-P1H01, R3G10-P1H08, R3G10-P2C04, R The sequences are 3G10-P2G11, R3G10-P3E04, R3G10-P4A02, R3G10-P4C05, R3G10-P4D04, R3G10-P4D10, R3G10-P4E07, R3G10-P4E12, R3G10-P4G06, R3G10-P5A08, or R3G10-P5C08. Table 8 shows the VH and VL sequences of identified scFvs that specifically bind to A*01:01_ASSLPTTMNY (SEQ ID NO:260). For clarity, each identified scFv hit is named with a clone name, and each row contains the VH and VL sequences for that specific clone name. For example, the scFv identified by clone name R3G10-P1A07 contains the VH sequence EVQLLESGGGLVKPGGSLRLSCAASGFTFSSYWMSWVRQAPGKGLEWVSGISARSGRTYYADSVKGRFTISRDDSKNTLYLQMNSLKTEDTAVYYCARDQDTIFGVVITWFDPWGQGTLVTVSS (SEQ ID NO:290) and the VL sequence DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYFTTPYTFGQGTKLEIK (SEQ ID NO:307).

[0422] Antibody specific to A*02:01LLASSILCA (SEQ ID NO:322)(G7)

[0423] In some respects, this document provides an ABP comprising an antibody or an antigen-binding fragment thereof that specifically binds to an HLA-peptide target, wherein the HLA class I molecule of the HLA-peptide target is HLA subtype A*02:01, and the HLA-restricted peptide of the HLA-peptide target comprises the sequence LLASSILCA (SEQ ID NO:322) (“G7”), is composed of or substantially composed of the sequence said.

[0424] Sequence of G7 specific antibody

[0425] As described in further detail, an ABP specific to A*02:01_LLASSILCA (SEQ ID NO:322) may comprise one or more sequences.

[0426] CDR

[0427] ABP specific to A*02:01_LLASSILCA (SEQ ID NO:322) may contain one or more antibody complementarity-determining region (CDR) sequences, for example, three heavy chain CDRs (CDR-H1, CDR-H2, CDR-H3) and three light chain CDRs (CDR-L1, CDR-L2, CDR-L3).

[0428] ABP specific to A*02:01_LLASSILCA (SEQ ID NO:322) may contain a CDR-H3 sequence. The CDR-H3 sequence may be selected from CARDGYDFWSGYTSDDYW (SEQ ID NO:323), CASDYGDYR (SEQ ID NO:324), CADLMTTVVTPGDYGMDVW (SEQ ID NO:325), CARQDGGAFAFDIW (SEQ ID NO:326), CARELGYYYGMDVW (SEQ ID NO:327), CAALIFGVPLLPYGMDVW (SEQ ID NO:328), CAKDLATVGEPYYYYGMDVW (SEQ ID NO:329), and CARLWFGELHYYYYYGMDVW (SEQ ID NO:330).

[0429] ABP specific to A*02:01_LLASSILCA (SEQ ID NO:322) may contain a CDR-L3 sequence. The CDR-L3 sequence may be selected from CHHYGRSHTF (SEQ ID NO:331), CQQANAFPPTF (SEQ ID NO:332), CQQYYSIPLTF (SEQ ID NO:333), CQQSYSTPPTF (SEQ ID NO:334), CQQSYSFPYTF (SEQ ID NO:335), CMQALQTPLTF (SEQ ID NO:31), CQQGNTFPLTF (SEQ ID NO:336), and CMQGSHWPPSF (SEQ ID NO:337).

[0430] An ABP specific to A*02:01_LLASSILCA (SEQ ID NO:322) may contain a specific heavy chain CDR3 (CDR-H3) sequence and a specific light chain CDR3 (CDR-L3) sequence. In some embodiments, the ABP contains CDR-H3 and CDR-L3 from scFvs named G7R3-P1C6, G7R3-P1G10, 1-G7R3-P1B4, 2-G7R4-P2C2, 3-G7R4-P1A3, 4-G7R4-B5-P2E9, 5-G7R4-B10-P1F8, or B7 (G7R3-P3A9). Table 36 shows the CDR sequences of identified scFvs that specifically bind to A*02:01_LLASSILCA (SEQ ID NO:322). For clarity, each identified scFv match is named with a clone name, and each line contains the CDR sequence for that specific clone name. For example, the scFv identified by the clone name G7R3-P1C6 contains the heavy chain CDR3 sequence CARDGYDFWSGYTSDDYW (SEQ ID NO:323) and the light chain CDR3 sequence CHHYGRSHTF (SEQ ID NO:331).

[0431] An ABP specific to A*02:01_LLASSILCA (SEQ ID NO:322) may contain all six CDRs from scFv, which are named G7R3-P1C6, G7R3-P1G10, 1-G7R3-P1B4, 2-G7R4-P2C2, 3-G7R4-P1A3, 4-G7R4-B5-P2E9, 5-G7R4-B10-P1F8 or B7 (G7R3-P3A9).

[0432] VL

[0433] An ABP specific to *02:01_LLASSILCA (SEQ ID NO: 322) may contain a VL sequence. The VL sequence may be selected from EIVMTQSPATLSVSPGERATLSCRASQSVSSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCHHYGRSHTFGQGTKVEIK (SEQ ID NO: 346), DIQMTQSPSSLSASVGDRVTITCRASQDIRNDLGWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANAFPPTFGQGTKVEIK (SEQ ID NO: 347), DIVMTQSPDSLAVSLGERATINCKSSQSVFYSSNNKNQLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSIPLTFGQGTKLEIK (SEQ ID NO: 348), DIQMTQSPSSLSASVGDRVTITCQASQDIFKYLNWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPTFGQGTRLEIK (SEQ ID NO: 349), DIQMTQSPSSLSASVGDRVTITCRASQSISTWLAWYQQKPGKAPKLLIYYASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSFPYTFGQGTKVEIK (SEQ ID NO: 350), DIVMTQSPLSLPVTPGEPASISCSSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTPLTFGGGTKVEIK (SEQ ID NO: 351), DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYSASNLRSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGNTFPLTFGQGTKVEIK (SEQ IDNO: 352) and DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGSHWPPSFGQGTRLEIK (SEQ ID NO: 353).

[0434] VH

[0435] An ABP specific for *02:01_LLASSILCA (SEQ ID NO:322) may comprise a VH sequence. The VH sequence may be selected from QVQLVQSGAEVKKPGASVKVSCKASGGTFSNYGISWVRQAPGQGLEWMGIINPGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGYDFWSGYTSDDYWGQGTLVTVSS (SEQ ID NO:338), EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMHWVRQAPGKGLEWVSGISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASDYGDYRGQGTLVTVSS (SEQ ID NO:339), QVQLVQSGAEVKKPGASVKVSCKASGYTFSNYYIHWVRQAPGQGLEWMGWLNPNSGNTGYAQRFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDLMTTVVTPGDYGMDVWGQGTTVTVSS (SEQ ID NO:340), QVQLVQSGAEVKKPGASMKVSCKASGYTFTTDGISWVRQAPGQGLEWMGRIYPHSGYTEYAKKFKGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARQDGGAFAFDIWGQGTMVTVSS (SEQ ID NO:341), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSQYMHWVRQAPGQGLEWMGWISPNNGDTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARELGYYYGMDVWGQGTTVTVSS (SEQ ID NO:342), QVQLVQSGAEVKKPGSSVKVSCKASRYTFTSYDINWVRQAPGQGLEWMGRIIPMLNIANYAPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARALIFGVPLLPYGMDVWGQGTTVTVSS (SEQ IDNO: 343), EVQLLQSGGGLVQPGGSLRLSCAASGFTFSSSWMHWVRQAPGKGLEWVSFISTSSGYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDLATVGEPYYYYGMDVWGQGTTVTVSS (SEQ ID NO: 344) and QVQLVQSGAEVKKPGSSVKVSCKASGDTFNTYALSWVRQAPGQGLEWMGWMNPNSGNAGYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARLWFGELHYYYYYGMDVWGQGTMVTVSS (SEQ ID NO: 345).

[0436] VH-VL combination

[0437] An ABP specific to A*02:01_LLASSILCA (SEQ ID NO:322) may contain a specific VH sequence and a specific VL sequence. In some embodiments, an ABP specific to A*02:01_LLASSILCA (SEQ ID NO:322) contains a VH sequence and a VL sequence from an scFv named G7R3-P1C6, G7R3-P1G10, 1-G7R3-P1B4, 2-G7R4-P2C2, 3-G7R4-P1A3, 4-G7R4-B5-P2E9, 5-G7R4-B10-P1F8, or B7 (G7R3-P3A9). Table 35 shows the VH and VL sequences of identified scFvs that specifically bind to A*02:01_LLASSILCA (SEQ ID NO:322). For clarity, each identified scFv match is named with a clone name, and each line contains the VH and VL sequences for that specific clone name. For example, the scFv identified by clone name G7R3-P1C6 contains the VH sequence QVQLVQSGAEVKKPGASVKVSCKASGGTFSNYGISWVRQAPGQGLEWMGIINPGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGYDFWSGYTSDDYWGQGTLVTVSS (SEQ ID NO:338) and the VL sequence EIVMTQSPATLSVSPGERATLSCRASQSVSSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCHHYGRSHTFGQGTKVEIK (SEQ ID NO:346).

[0438] Antibodies specific to A*01:01_NTDNNLAVY (SEQ ID NO:73)(G2)

[0439] In some respects, this document provides an ABP comprising an antibody or an antigen-binding fragment thereof that specifically binds to an HLA-PEPTIDE target, wherein the HLA class I molecule of the HLA-peptide target is HLA subtype A*01:01, and the HLA-restricted peptide of the HLA-peptide target comprises the sequence NTDNNLAVY (SEQ ID NO:73) (“G2”), is composed of, or is substantially composed of the sequence said.

[0440] Sequence of G2-specific antibody

[0441] As described in further detail, an ABP specific to A*01:01_NTDNNLAVY (SEQ ID NO:73) may comprise one or more sequences.

[0442] CDR

[0443] ABP specific to A*01:01_NTDNNLAVY (SEQ ID NO:73) may contain one or more antibody complementarity-determining region (CDR) sequences, for example, it may contain three heavy chain CDRs (CDR-H1, CDR-H2, CDR-H3) and three light chain CDRs (CDR-L1, CDR-L2, CDR-L3).

[0444] An ABP specific for A*01:01_NTDNNLAVY (SEQ ID NO:73) may comprise a CDR-H3 sequence. The CDR-H3 sequence may be selected from CAATEWLGVW (SEQ ID NO:74), CARANWLDYW (SEQ ID NO:75), CARANWLDYW (SEQ ID NO:75), CARDWVLDYW (SEQ ID NO:76), CARGEWLDYW (SEQ ID NO:77), CARGWELGYW (SEQ ID NO:78), CARDFVGYDDW (SEQ ID NO:79), CARDYGDLDYW (SEQ ID NO:80), CARGSYGMDVW (SEQ ID NO:81), CARDGYSGLDVW (SEQ ID NO:82), CARDSGVGMDVW (SEQ ID NO:83), CARDGVAVASDYW (SEQ ID NO:84), CARGVNVDDFDYW (SEQ ID NO:85), CARGDYTGNWYFDLW (SEQ ID NO:86), CARANWLDYW (SEQ ID NO:75), CARDQFYGGNSGGHDYW (SEQ ID NO:87), CAREEDYW (SEQ ID NO:88), CARGDWFDPW (SEQ ID NO:89), CARGDWFDPW (SEQ ID NO:89), CARGEWFDPW (SEQ ID NO:90), CARSDWFDPW (SEQ ID NO:91), CARDSGSYFDYW (SEQ ID NO:92), CARDYGGYVDYW (SEQ ID NO:93), CAREGPAALDVW (SEQ ID NO:94), CARERRSGMDVW (SEQ ID NO:95), CARVLQEGMDVW (SEQ ID NO:96), CASERELPFDIW (SEQ ID NO:97), CAKGGGGYGMDVW (SEQ ID NO:98), CAAMGIAVAGGMDVW (SEQ ID NO:99), CARNWNLDYW (SEQ ID NO:100), CATYDDGMDVW (SEQ ID NO:101), CARGGGGALDYW (SEQ ID NO:102), CALSGNYYGMDVW (SEQ ID NO:103), CARGNPWELRLDYW (SEQ ID NO:104), and CARDKNYYGMDVW (SEQ ID NO:105).

[0445] An ABP specific for A*01:01_NTDNNLAVY (SEQ ID NO:73) may comprise a CDR-L3 sequence. The CDR-L3 sequence may be selected from CQQSYNTPYTF (SEQ ID NO:106), CQQSYSTPYTF (SEQ ID NO:107), CQQSYSTPYSF (SEQ ID NO:108), CQQSYSTPFTF (SEQ ID NO:109), CQQSYGVPYTF (SEQ ID NO:110), CQQSYSAPYTF (SEQ ID NO:111), CQQSYSAPYTF (SEQ ID NO:111), CQQSYSAPYSF (SEQ ID NO:112), CQQSYSTPYTF (SEQ ID NO:107), CQQSYSVPYSF (SEQ ID NO:113), CQQSYSAPYTF (SEQ IDNO:111), CQQSYSVPYSF (SEQ ID NO:113), CQQSYSTPQTF (SEQ ID NO:114), CQQLDSYPFTF(SEQID NO:115), CQQSYSSPYTF (SEQ ID NO:116), CQQSYSTPLTF (SEQ ID NO:30), CQQSYSTPYSF (SEQ ID NO:108), CQQSYSTPYTF (SEQ ID NO:107), CQQSYSTPYTF (SEQ ID NO:107), CQQSYSTPFTF (SEQ ID NO:109), CQQSYSTPTF (SEQ ID NO:117), CQQTYAIPLTF (SEQ ID NO:118), CQQSYSTPYTF (SEQ ID NO:107), CQQSYIAPFTF (SEQ ID NO:119), CQQSYSIPLTF (SEQ IDNO:25), CQQSYSNPTF (SEQ ID NO:120), CQQSYSTPYSF (SEQ ID NO:108), CQQSYSDQWTF(SEQID NO:121), CQQSYLPPYSF (SEQ ID NO:122), CQQSYSSPYTF (SEQ ID NO:116), CQQSYTTPWTF (SEQ ID NO:123), CQQSYLPPYSF (SEQ ID NO:122), CQEGITYTF (SEQ ID NO:124), CQQYYSYPFTF (SEQ ID NO:125) and CQHYGYSPVTF (SEQ ID NO:126).

[0446] An ABP specific to A*01:01_NTDNNLAVY (SEQ ID NO:73) may contain a specific heavy chain CDR3 (CDR-H3) sequence and a specific light chain CDR3 (CDR-L3) sequence. In some embodiments, the ABP contains CDR-H3 and CDR-L3 from scFvs named G2-P2E07, G2-P2E03, G2-P2A11, G2-P2C06, G2-P1G01, G2-P1C02, G2-P1H01, G2-P1B12, G2-P1B06, G2-P2H10, G2-P1H10, G2-P2C11, G2-P1C09, G2-P1A10, G2-P1B10, G2-P1D07, G2-P1E05, G2-P1D03, G2-P1G12, G2-P2H11, G2-P1C03, G2-P1G07, G2-P1F12, G2-P1G03, G2-P2B08, G2-P2A10, G2-P2D04, G2-P1C06, G2-P2A09, G2-P1B08, G2-P1E03, G2-P2A03, G2-P2F01, G2-P1H11, or G2-P1D06. The identified CDR sequences of scFv that specifically bind to A*01:01_NTDNNLAVY (SEQ ID NO:73) are shown in Table 34. For clarity, each identified scFv match is named with a clone name, and each line contains the CDR sequence for that specific clone name. For example, the scFv identified by clone name G2-P2E07 contains the heavy chain CDR3 sequence CAATEWLGVW (SEQ ID NO:74) and the light chain CDR3 sequence CQQSYNTPYTF (SEQ ID NO:106).

[0447] An ABP specific to A*01:01_NTDNNLAVY (SEQ ID NO:73) may contain all six CDRs from scFv, which are named G2-P2E07, G2-P2E03, G2-P2A11, G2-P2C06, G2-P1G01, G2-P1C02, G2-P1H01, G2-P1B12, G2-P1B06, G2-P2H10, G2-P1H10, G2-P2C11, G2-P1C09, G2-P1A10, G2-P1B10, G... 2-P1D07, G2-P1E05, G2-P1D03, G2-P1G12, G2-P2H11, G2-P1C03, G2-P1G07, G2-P1F12, G2-P1G03, G2-P2B08, G2-P2A10, G2-P2D04, G2-P1C06, G2-P2A09, G2-P1B08, G2-P1E03, G2-P2A03, G2-P2F01, G2-P1H11 or G2-P1D06.

[0448] VL

[0449] The ABP specific for A*01:01_NTDNNLAVY (SEQ ID NO:73) contains a VL sequence. The VL sequence can be selected from DIQMTQSPSSLSASVGDRVTITCRASQSISTWLAWYQQKPGKAPKLLIYAASSLRSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYNTPYTFGQGTKLEIK (SEQ ID NO:163), DIQMTQSPSSLSASVGDRVTITCRASQSISRWLAWYQQKPGKAPKLLIYAASTVQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTKLEIK (SEQ ID NO:164), DIQMTQSPSSLSASVGDRVTITCRASQDISRWLAWYQQKPGKAPKLLIYAASRLQAGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYSFGQGTKLEIK (SEQ ID NO:165), DIQMTQSPSSLSASVGDRVTITCRASQTISSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKVDIK (SEQ ID NO:166), DIQMTQSPSSLSASVGDRVTITCRASQTISSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYGVPYTFGQGTKVEIK (SEQID NO:167), DIQMTQSPSSLSASVGDRVTITCRASQSISNWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSAPYTFGPGTKVDIK (SEQ ID NO:168), DIQMTQSPSSLSASVGDRVTITCRASQSVGNWLAWYQQKPGKAPKLLIYGASSLQTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSAPYTFGQGTKVEIK (SEQ IDNO:169)、DIQMTQSPSSLSASVGDRVTITCRASQNIGNWLAWYQQKPGKAPKLLIYAASTLQTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSAPYSFGQGTKLEIK(SEQ ID NO:170)、DIQMTQSPSSLSASVGDRVTITCRASQSISRWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQTGKLEIK(SEQ ID NO:170) ID NO:171)、DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAKPLLIYGASLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSVPYSFGQGTKLEIK(SEQ ID NO:172)、DIQMTQSPSSLSASVGDRVTITCRASQSISKWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSAPYTFGQGTKVEIK(SEQ ID NO:173), DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSVPYSFGQGTKLEIK(SEQ ID NO:174), DIQMTQSPSSLSASVGDRVTITCRASQTISNYLNWYQQKPGKAPKLLIYAASNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPQTFGQGTKVEIK(SEQ ID NO:175), DIQMTQSPSSLSASVGDRVTITCRASRDIGRAVGWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQLDSYPFTFGPGTKVDIK(SEQ ID NO:176), DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSSPYTFGPGTKVDIK(SEQ ID NO:177)NO:177)、DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKLEIK(SEQ ID NO:178)、DIQMTQSPSSLSASVGDRVTITCRASQSIGRWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYSFGQGTKVEIK(SEQ ID NO:179)、DIQMTQSPSSLSASVGDRVTITCRASQSISTWLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFAQGTKLEIK(SEQ ID NO:180)、DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYGASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTKLEIK(SEQ ID NO:181)、DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKVDIK(SEQ ID NO:182)、DIQMTQSPSSLSASVGDRVTITCRASQSVSNWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPTFGQGTKLEIK(SEQ ID NO:183)、DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTYAIPLTFGGGTKVEIK(SEQ ID NO:184)、DIQMTQSPSSLSASVGDRVTITCQASQDIGSWLAWYQQKPGKAPKLLIYATSSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTKLEIK(SEQ IDNO:185)、DIQMTQSPSSLSASVGDRVTITCRASQGISRWLAWYQQKPGKAPKLLIYAASTLQPGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYIAPFTFGPGTKVDIK(SEQ ID NO:186)、DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASRLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSIPLTFGGGTKVEIK(SEQ ID NO:187)、DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYGVSSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSNPTFGQGTKVEIK(SEQID NO:188)、DIQMTQSPSSLSASVGDRVTITCRASQSISSWVAWYQQKPGKAPKLLIYGASNLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYSFGQGTKLEIK(SEQ ID NO:189)、DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSDQWTFGQGTKVEIK(SEQ ID NO:190)、DIQMTQSPSSLSASVGDRVTITCRASQSISRWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYLPPYSFGQGTKVEIK(SEQ ID NO:191)、DIQMTQSPSSLSASVGDRVTITCRASQSISNWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTYFTLTISSLQPEDFATYYCQQSYSSPYTFGQGTKLEIK(SEQ ID NO:192)、DIQMTQSPSSLSASVGDRVTITCRASQSISHYLNWYQQKPGKAPKLLIYGASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYTTPWTFGQGTRLEIK(SEQ IDNO: 193), DIQMTQSPSSLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYLPPYSFGQGTKLEIK (SEQ ID NO: 194), DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYGASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQEGITYTFGQGTKVEIK (SEQ ID NO: 195), DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYSYPFTFGPGTKVDIK (SEQ ID NO: 162) and EIVMTQSPATLSVSPGERATLSCRASQSVSRNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQHYGYSPVTFGQGTKLEIK (SEQ ID NO: 196).

[0450] VH

[0451] An ABP specific for A*01:01_NTDNNLAVY (SEQ ID NO:73) may comprise a VH sequence. The VH sequence may be selected from QVQLVQSGAEVKKPGASVKVSCKASGGTFSSATISWVRQAPGQGLEWMGWIYPNSGGTVYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAATEWLGVWGQGTTVTVSS (SEQ ID NO:128), EVQLLQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGWINPNSGGTISAPNFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARANWLDYWGQGTLVTVSS (SEQ ID NO:129), EVQLLESGAEVKKPGASVKVSCKASGYTFTTYDLAWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARANWLDYWGQGTLVTVSS (SEQ ID NO:130), QVQLVQSGAEVKKPGASVKVSCKSSGYSFDSYVVNWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDWVLDYWGQGTLVTVSS (SEQ ID NO:131), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWMNPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGEWLDYWGQGTLVTVSS (SEQ ID NO:132), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGWELGYWGQGTLVTVSS (SEQ ID NO:133), QVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTINWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDFVGYDDWGQGTLVTVSS (SEQ IDNO:134)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGITWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDYGDLDYWGQGTLVTVSS(SEQ ID NO:135)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSNYILSWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSYGMDVWGQGTTVTVSS(SEQ ID NO:136)、QVQLVQSGAEVKKPGASVKVSCKASGYSFTRYNMHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGYSGLDVWGKGTTVTVSS(SEQID NO:137)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGWINPNNGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDSGVGMDVWGQGTTVTVSS(SEQID NO:138)、QVQLVQSGAEVKKPGASVKVSCKASGGTFNNYAFSWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDGVAVASDYWGQGTLVTVSS(SEQ ID NO:139)、QVQLVQSGAEVKKPGASVKVSCKASGYTFSSYNMHWVRQAPGQGLEWMGWINGNTGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGVNVDDFDYWGQGTLVTVSS(SEQ ID NO:140)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSSYAFSWVRQAPGQGLEWMGWINPDTGYTRYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGDYTGNWYFDLWGRGTLVTVSS(SEQ IDNO:141)、EVQLLESGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWINPYSGGTNYAQKLQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARANWLDYWGQGTLVTVSS(SEQ ID NO:142)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWISAYNGYTNYAQNLQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDQFYGGNSGGHDYWGQGTLVTVSS(SEQ ID NO:143)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYNMHWVRQAPGQGLEWMGWMNPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAREEDYWGQGTLVTVSS(SEQ ID NO:144)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTINWVRQAPGQGLEWMGWINPNSGGANYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGDWFDPWGQGTLVTVSS(SEQ ID NO:145)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQGLEWMGWISPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGDWFDPWGQGTLVTVSS(SEQ ID NO:146)、QVQLVQSGAEVKKPGASVKVSCKASGYTFSDYYVHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGEWFDPWGQGTLVTVSS(SEQ ID NO:147)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTTYYMHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARSDWFDPWGQGTLVTVSS(SEQ IDNO:148)、QVQLVQSGAEVKKPGASVKVSCKASGGTFSNYAINWVRQAPGQGLEWMGWISPYSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDSGSYFDYWGQGTLVTVSS(SEQ ID NO:149)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYYMHWVRQAPGQGLEWMGWIYPNTGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDYGGYVDYWGQGTLVTVSS(SEQID NO:150)、EVQLLESGAEVKKPGASVKVSCKASGYTFTSYAMNWVRQAPGQGLEWMGWMNPNSGGTKYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAREGPAALDVWGQGTLVTVSS(SEQ ID NO:151)、QVQLVQSGAEVKKPGASVKVSCKASGYTLTSHLIHWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARERRSGMDVWGQGTTVTVSS(SEQ ID NO:152)、EVQLLESGAEVKKPGASVKVSCKASGYSFTDYIVHWVRQAPGQGLEWMGWINPYSGGTKYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARVLQEGMDVWGQGTLVTVSS(SEQ ID NO:153)、QVQLVQSGAEVKKPGASVKVSCKASGYTFSNFLINWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCASERELPFDIWGQGTMVTVSS(SEQID NO:154)、QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYQMFWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAKGGGGYGMDVWGQGTTVTVSS(SEQ IDNO:155), QVQLVQSGAEVKKPGASVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGWINPNSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAAMGIAVAGGMDVWGQGTLVTVSS (SEQ ID NO:156), QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYHMHWVRQAPGQGLEWMGWIHPDSGGTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARNWNLDYWGQGTLVTVSS (SEQ ID NO:157), QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWMNPNSGNTGYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCATYDDGMDVWGQGTTVTVSS (SEQ ID NO:158), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYTVNWVRQAPGQGLEWMGWINPNSGGTKYAQNFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGGGGALDYWGQGTLVTVSS (SEQ ID NO:159), QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGMINPRDDTTDYARDFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCALSGNYYGMDVWGQGTTVTVSS (SEQ ID NO:160), QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYMHWVRQAPGQGLEWMGMINPSGGGTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGNPWELRLDYWGQGTLVTVSS (SEQ ID NO:127) and QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSQYMHWVRQAPGQGLEWMGRIIPLLGIVNYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDKNYYGMDVWGQGTTVTVSS (SEQ ID NO:161).

[0452] VH-VL combination

[0453] ABP specific to A*01:01_NTDNNLAVY (SEQ ID NO:73) comprises a specific VH sequence and a specific VL sequence. In some embodiments, ABP specific to A*01:01_NTDNNLAVY (SEQ ID NO:73) comprises a VH sequence and a VL sequence from an scFv named G2-P2E07, G2-P2E03, G2-P2A11, G2-P2C06, G2-P1G01, G2-P1C02, G2-P1H01, G2-P1B12, G2-P1B06, G2-P2H10, G2-P1H10, G2-P2C11, G2-P1C09, G2-P1A10, G2-P1B10, G2-P1D07, G2-P1E05, G2-P1D03, G2-P1G12, G2-P2H11, G2-P1C03, G2-P1G07, G2-P1F12, G2-P1G03, G2-P2B08, G2-P2A10, G2-P2D04, G2-P1C06, G2-P2A09, G2-P1B08, G2-P1E03, G2-P2A03, G2-P2F01, G2-P1H11, or G2-P1D06. The VH and VL sequences of identified scFvs that specifically bind to A*01:01_NTDNNLAVY (SEQ ID NO:73) are shown in Table 33. For clarity, each identified scFv match is named with a clone name, and each line contains the CDR sequence for that specific clone name. For example, the scFv identified by clone name G2-P2E07 contains the VH sequence QVQLVQSGAEVKKPGASVKVSCKASGGTFSSATISWVRQAPGQGLEWMGWIYPNSGGTVYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCAATEWLGVWGQGTTVTVSSAS (SEQ ID NO:365) and the VL sequence DIQMTQSPSSLSASVGDRVTITCRASQSISTWLAWYQQKPGKAPKLLIYAASSLRSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYNTPYTFGQGTKLEIK (SEQ ID NO:163).

[0454] receptor

[0455] In the provided ABPs, such as the HLA-peptide ABP, the receptor is used. Receptors may include antigen receptors that specifically bind to the HLA-peptide targets disclosed herein, and other chimeric receptors. The receptor may be a T-cell receptor (TCR). The receptor may be a chimeric antigen receptor (CAR).

[0456] TCRs can be soluble or membrane-bound. Among antigen receptors are functional non-TCR antigen receptors, such as chimeric antigen receptors (CARs). Cells expressing the receptors and their use in adoptive cell therapies, such as those treating diseases and conditions associated with HLA-peptide expression, including cancer, are also provided.

[0457] Exemplary antigen receptors (including CARs) and methods for engineering and introducing such receptors into cells include those described in the following documents: for example, International Patent Application Publications Nos. WO200014257, WO2013126726, WO2012 / 129514, WO2014031687, WO2013 / 166321, WO2013 / 071154, and WO2013 / 123061; and U.S. Patent Application Publications Nos. US2002131960, US2013287748, and US20130. 149337; U.S. Patents 6,451,995, 7,446,190, 8,252,592, 8,339,645, 8,398,282, 7,446,179, 6,410,319, 7,070,995, 7,265,209, 7,354,762, 7,446,191, 8,324,353 and 8,479,118; and European Patent Application EP2537416, and / or those described in the following documents: Sadelain et al., Cancer Discov. April 2013; 3(4):388-398; Davila et al. (2013) PLoS ONE 8(4):e61338; Turtle et al., Curr. Opin. Immunol., October 2012; 24(5):633-39; Wu et al., Cancer, March 2012 18(2):160-75. In some respects, antigen receptors include the CARs described in U.S. Patent No. 7,446,190 and those described in International Patent Application Publication No. WO / 2014055668A1. Exemplary CARs include any of the CARs described in the aforementioned publications, such as WO2014031687, U.S. Patent No. 8,339,645, U.S. Patent No. 7,446,179, US 2013 / 0149337, U.S. Patent No. 7,446,190, U.S. Patent No. 8,389,282, etc., wherein the antigen-binding portion (e.g., scFv) is replaced by an antibody (e.g., the antibody provided herein).

[0458] Chimeric receptors include chimeric antigen receptors (CARs). Chimeric receptors, such as CARs, generally include an extracellular antigen-binding domain comprising, being, or contained within one of the provided anti-HLA-peptide ABPs (such as anti-HLA-peptide antibodies). Therefore, the extracellular portion of a chimeric receptor (e.g., a CAR) typically contains one or more HLA-peptide-ABPs, such as one or more antigen-binding fragments, domains, or portions, or one or more antibody variable domains, and / or antibody molecules (as described herein). In some embodiments, the CAR contains an HLA-peptide-binding portion or a portion of an ABP (e.g., an antibody) molecule, such as a variable heavy (VH) chain region and / or a variable light (VL) chain region of an antibody, for example, an scFv antibody fragment.

[0459] TCR

[0460] On the one hand, the ABPs provided herein, such as ABPs that specifically bind to the HLA-peptide targets disclosed herein, contain T-cell receptors (TCRs). TCRs can be isolated and purified.

[0461] In most T cells, the TCR is a heterodimeric polypeptide with alpha (α) and beta (β) chains encoded by TRA and TRB, respectively. The α chain typically contains an α variable region encoded by TRAV, an α linker region encoded by TRAJ, and an α constant region encoded by TRAC. The β chain typically contains a β variable region encoded by TRBV, a β diversity region encoded by TRBD, a β linker region encoded by TRBJ, and a β constant region encoded by TRBC. The TCR-α chain is generated via VJ recombination, while the β chain receptor is generated via V(D)J recombination. Additional diversity in the TCR arises from linker diversity. Several bases can be deleted and several bases added (called N and P nucleotides) at each linker. In most T cells, the TCR contains both γ and δ chains. The TCR γ chain is generated through VJ recombination, while the TCR δ chain is generated through V(D)J recombination (Kenneth Murphy, Paul Travers, and Mark Walport, Janeway's Immunology 7th ed., Garland Science, 2007, incorporated herein by reference in its entirety). The antigen-binding site of a TCR typically contains six complementarity-determining regions (CDRs). The α chain contributes three CDRs: αCDR1, αCDR2, and αCDR3. The β chain also provides three CDRs: βCDR1, βCDR2, and βCDR3. αCDR3 and βCDR3 are the regions most affected by V(D)J recombination and are responsible for most variations in the TCR repertoire.

[0462] TCRs can specifically recognize HLA-peptide targets, such as those disclosed in Tables A, A1, or A2; therefore, a TCR can be an ABP that specifically binds to HLA-peptides. TCRs can be soluble, such as antibodies similar to those secreted by B cells. TCRs can also be membrane-bound, for example, binding to cells such as T cells or natural killer (NK) cells. Therefore, TCRs can be used in the context of soluble antibodies and / or membrane-bound CARs.

[0463] Any TCR disclosed herein may include an α-variable region, an α-connection region, optionally an α-constant region, a β-variable region, optionally a β-diversity region, a β-connection region, and optionally a β-constant region.

[0464] In some implementations, the TCR or CAR is a recombinant TCR or CAR. A recombinant TCR or CAR may contain any TCR identified herein, but may contain one or more modifications. Exemplary modifications, such as amino acid substitutions, are described herein. The amino acid substitutions described herein are referred to in the IMGT nomenclature and the amino acid numbers available at www.imgt.org.

[0465] Recombinant TCRs or CARs can be human TCRs or CARs containing complete human sequences, such as natural human sequences. Recombinant TCRs or CARs may retain their natural human variable domain sequences but contain modifications to the α-constant region, β-constant region, or both. Such modifications to the TCR constant region can, for example, drive preferential pairing of exogenous TCR strands to improve TCR assembly and expression in TCR gene therapy.

[0466] In some implementations, the α and β constant regions are modified by replacing the mouse constant region sequence with a whole human constant region sequence. This “mouse-derived” TCR and its preparation method are described in Cancer Res. 2006 Sep 1; 66(17):8878-86, which is incorporated herein by reference in its entirety.

[0467] In some embodiments, the α and β constant regions are modified by substituting one or more amino acids in the human TCRα constant (TRAC) region, TCRβ constant (TRBC) region, or TRAC and TRAB regions, i.e., replacing human residues with mouse residues ((human → mouse amino acid exchange)). One or more amino acid substitutions in the TRAC region may 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. One or more amino acid substitutions in the human TRBC region may 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 2010 1 June, 184(11) 6223-6231, which is incorporated herein by reference in its entirety.

[0468] In some implementations, human TRAC contains an Asp substitution at residue 210, while human TRBC contains a Lys substitution at residue 134. This substitution can promote the formation of salt bridges between the α and β chains and disulfide bonds between the TCR chains. These targeted substitutions are described in J Immunol 2010 1 June; 184(11) 6232-6241, which is incorporated herein by reference in its entirety.

[0469] In some implementations, the human TRAC and human TRBC regions are modified to contain introduced cysteine residues, which can improve the preferential pairing of exogenous TCR chains by forming additional disulfide bonds. For example, the human TRAC may contain a Cys substitution at residue 48, and the human TRBC may contain a Cys substitution at residue 57, as described in the following references: Cancer Research, April 15, 2007; 67(8):3898-903 and Blood, March 15, 2007; 109(6):2331-8; which are incorporated herein by reference in their entirety.

[0470] Recombinant TCRs or CARs may contain additional modifications to the α and β chains.

[0471] In some implementations, the α and β chains are modified by linking the extracellular domains of the α and β chains to the intact human CD3ζ ((CD3-zeta)) molecule. Such modifications are described in the following references: J Immunol 1 June 2008, 180(11) 7736-7746; Gene Ther. Aug 2000; 7(16): 1369-77; and The Open Gene Therapy Journal, 2011, 4: 11-22 (the references are hereby incorporated in their entirety by reference).

[0472] In some implementations, the α chain is modified by introducing hydrophobic amino acid substitutions into the transmembrane region of the α chain, as described in JImmunol 1 June 2012; 188(11)5538-5546; Gene Ther. 7(16):1369-77; and The Open Gene Therapy Journal, 2011, 4:11-22 (the entire references are hereby incorporated by reference).

[0473] The α or β chain can be modified by altering any N-glycosylation site in the amino acid sequence, as described in J Exp Med. 2009 Feb 16; 206(2):463–475 (which is hereby incorporated in its entirety by reference).

[0474] The α and β chains may each contain a dimerizing domain, such as a heterodimerizing domain. Such heterodimerizing domains are known in the art to be leucine zippers (5H3 domains), hydrophobic proline-rich reverse domains, or other similar forms. In one instance, the α and β chains can be modified by introducing 30-merging segments into the carboxyl ends of the α and β extracellular domains, wherein said segments selectively associate to form stable leucine zippers. This modification is described in: PNAS November 22, 1994, 91(24)11408-11412; https: / / doi.org / 10.1073 / pnas.91.24.11408; which is incorporated herein by reference in its entirety.

[0475] The TCRs identified in this paper may be modified to include mutations that result in increased affinity or half-life, such as those described in WO2012 / 013913, which are incorporated herein by reference in their entirety.

[0476] Recombinant TCRs or CARs can be single-stranded TCRs (scTCRs). Such scTCRs may include an α-strand variable region sequence fused to the N-terminus of the extracellular sequence of the α-strand constant region of the TCR, a TCR β-strand variable region fused to the N-terminus of the extracellular sequence of the β-strand constant region of the TCR, and a linker sequence connecting the C-terminus of the α-strand to the N-terminus of the β-strand, or vice versa. In some embodiments, the extracellular sequences of the α-strand and β-strand constant regions of the scTCR are linked by disulfide bonds. In some embodiments, the length of the linker sequence and the position of the disulfide bonds cause the variable region sequences of the α-strand and β-strand to be substantially oriented to each other as in the native αβT cell receptor. An exemplary scTCR is described in U.S. Patent No. 7,569,664, which is incorporated herein by reference in its entirety.

[0477] In some cases, the variable region of the scTCR can be covalently linked via a short peptide linker, as described in Gene Therapy, Vol. 7, pp. 1369–1377 (2000). The short peptide linker may be serine-rich or glycine-rich. For example, the linker may be (Gly4Ser)3 (SEQ ID NO: 366), as described in Cancer Gene Therapy (2004) 11, 487–496 (which is incorporated herein by reference in its entirety).

[0478] Recombinant TCRs or their antigen-binding fragments can be expressed as fusion proteins. For example, a TCR or its antigen-binding fragment can be fused with a toxin. Such a fusion protein is described in Cancer Res. 2002, March 15; 62(6):1757-60. A TCR or its antigen-binding fragment can also be fused with the Fc region of an antibody. Such a fusion protein is described in J Immunol. 2017, May 1, 198((1 Supplement))120.9.

[0479] In some embodiments, recombinant receptors such as TCRs or CARs (e.g., their antibody portions) also include a spacer, which may be or contain at least a portion of an immunoglobulin constant region or a variant or modified form thereof, such as a hinge region, for example, the IgG4 hinge region and / or CH1 / CL and / or Fc region. In some embodiments, the constant region or portion is human IgG, such as IgG4 or IgG1. In some aspects, a portion of the constant region serves as a spacer between an antigen recognition component (e.g., scFv) and a transmembrane domain. The length of the spacer can provide increased cellular reactivity after antigen binding compared to the absence of a spacer. In some instances, the length of the spacer is about 12 amino acids, or not more than 12 amino acids. Exemplary spacers comprise those having at least about 10 to 229 amino acids, about 10 to 200 amino acids, about 10 to 175 amino acids, about 10 to 150 amino acids, about 10 to 125 amino acids, about 10 to 100 amino acids, about 10 to 75 amino acids, about 10 to 50 amino acids, about 10 to 40 amino acids, about 10 to 30 amino acids, about 10 to 20 amino acids, or about 10 to 15 amino acids, and those comprising any integer between the endpoints of any of the listed ranges. In some embodiments, the spacer region has about 12 or fewer amino acids, about 119 or fewer amino acids, or about 229 or fewer amino acids. Exemplary spacers comprise individual IgG4 hinges, IgG4 hinges connected to the CH2 and CH3 domains, or IgG4 hinges connected to the CH3 domain. Exemplary spacers include, but are not limited to, those described in the following documents: Hudecek et al. (2013) Clin. Cancer Res., 19:3153, or International Patent Application Publication No. WO2014031687. In some embodiments, the constant region or portion is IgD.

[0480] 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, in the case of a CAR, signaling components activated by an antigen-receptor complex (such as a TCR complex), and / or signaling via another cell surface receptor. Therefore, in some embodiments, an HLA-peptide-specific binding component (e.g., ABP, such as an antibody or TCR) is linked to one or more transmembrane and intracellular signaling domains. In some embodiments, the transmembrane domain is fused to an extracellular domain. In one embodiment, a transmembrane domain natively associated with one of the domains in the receptor (e.g., CAR) is used. In some cases, the transmembrane domain is selected or modified by amino acid substitution to prevent this domain from binding to transmembrane domains of the same or different surface membrane proteins, thereby minimizing interactions with other members of the receptor complex.

[0481] In some embodiments, the transmembrane domain is natural or synthetic. If of natural origin, in some aspects, the domain is derived from any membrane-binding or transmembrane protein. The transmembrane region comprises a transmembrane region (i.e., a transmembrane region containing at least these) derived from the α, β, or ζ chain of the T cell receptor, CD28, CD3ε, CD45, CD4, CD5, CDDS, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, and / or CD154. Alternatively, in some embodiments, the transmembrane domain is synthetic. In some aspects, the synthetic transmembrane domain primarily comprises hydrophobic residues such as leucine and valine. In some aspects, a triplet of phenylalanine, tryptophan, and valine is present at each end of the synthetic transmembrane domain. In some embodiments, the domain is linked by a linker, spacer, and / or transmembrane domain.

[0482] Intracellular signal transduction domains contain those that mimic or approximate signals transmitted through natural antigen receptors, signals transmitted through a combination of such receptors and co-stimulatory receptors, and / or signals transmitted solely through co-stimulatory receptors. In some embodiments, short oligonucleotide or polypeptide linkers are present, such as linkers between 2 and 10 amino acids in length, like linkers containing glycine and serine, such as glycine-serine duplexes, and form a connection between the transmembrane domain and the cytoplasmic signal transduction domain of the receptor.

[0483] The receptor, such as a TCR or CAR, may contain at least one or more intracellular signaling components. In some embodiments, the receptor contains an intracellular component of the TCR complex, such as the TCR CD3 chain, for example, the CD3ζ chain, which mediates T cell activation and cytotoxicity. Therefore, in some aspects, an HLA-peptide-binding ABP (e.g., an antibody) is linked to one or more cell signaling modules. In some embodiments, the cell signaling module contains a CD3 transmembrane domain, a CD3 intracellular signaling domain, and / or other CD transmembrane domains. In some embodiments, the receptor (e.g., CAR) further contains portions of one or more additional molecules, such as Fc receptor-γ, CD8, CD4, CD25, or CD16. For example, in some aspects, the CAR contains a chimeric molecule between CD3ζ or Fc receptor-γ and CD8, CD4, CD25, or CD16.

[0484] In some embodiments, once the TCR or CAR is attached, the cytoplasmic or intracellular signaling domain of the receptor activates at least one of normal effector function or immune cell (e.g., engineered T cells expressing the receptor) response. For example, in some cases, the receptor induces T cell function, such as cytolytic activity or T helper activity, such as the secretion of cytokines or other factors. In some embodiments, for example, if the intracellular signaling domain of the antigen receptor component transduces effector signaling, the complete immunostimulatory chain is replaced with a truncated portion of the intracellular signaling domain of the antigen receptor component or a co-stimulatory molecule. In some embodiments, one or more intracellular signaling domains contain a cytoplasmic sequence of a T cell receptor (TCR) and, in some respects, also include those co-receptors that, in their natural environment, synergize with such a receptor to initiate signal transduction upon antigen receptor binding, and / or any derivatives or variants of such molecules, and / or any synthetic sequence having the same function.

[0485] In the case of a natural TCR, full activation typically requires not only signal transduction via the TCR but also co-stimulatory signals. Therefore, in some embodiments, components for generating secondary or co-stimulatory signals are also included in the receptor to facilitate full activation. In other embodiments, the receptor does not contain components for generating co-stimulatory signals. In some aspects, additional receptors are expressed in the same cell and provide components for generating secondary or co-stimulatory signals.

[0486] In some respects, T cell activation is described as being mediated by two classes of cytoplasmic signaling sequences: those that induce antigen-dependent primary activation via the TCR (primary cytoplasmic signaling sequence), and those that act in an antigen-independent manner to provide secondary or co-stimulatory signals (secondary cytoplasmic signaling sequences). In some respects, receptors contain one or both of these signaling components.

[0487] In some embodiments, the receptor contains a major cytoplasmic signaling sequence that regulates primary activation of the TCR complex. This major cytoplasmic signaling sequence, which functions in a stimulatory manner, may contain signaling motifs known as immune receptor tyrosine-based activation motifs or ITAMs. Examples of ITAM-containing major cytoplasmic signaling sequences include sequences derived from TCR or CD3ζ, FcRγ, FcRβ, CD3γ, CD3δ, CD3ε, CDS, CD22, CD79a, CD79b, and CD66d. In some embodiments, the cytoplasmic signaling molecule in the CAR contains a cytoplasmic signaling domain, a portion thereof, or a sequence derived from CD3ζ.

[0488] In some implementations, the receptor includes the signal transduction domain and / or transmembrane portion of a co-stimulatory receptor, such as CD28, 4-1BB, OX40, DAP10, and ICOS. In some aspects, the same receptor contains both an activating component and a co-stimulatory component.

[0489] In some embodiments, the activation domain is contained within a receptor, while the co-stimulatory component is provided by another receptor that recognizes a different antigen. In some embodiments, the receptor comprises both an activating or stimulating receptor and a co-stimulatory receptor expressed on the same cell (see WO2014 / 055668). In some aspects, the receptor targeting the HLA-peptide is a stimulatory or activating receptor. In other aspects, it is a co-stimulatory receptor. In some embodiments, the cell also contains an inhibitory receptor (e.g., iCAR, see Fedorov et al., Sci. Transl. Medicine, 5(215)(December 2013)), such as a receptor that recognizes antigens other than the HLA-peptide, thereby reducing or inhibiting the activation signal transmitted via the receptor targeting the HLA-peptide through the binding of the inhibitory receptor to its ligand, for example, to reduce off-target effects.

[0490] In some embodiments, the intracellular signaling domain includes a CD28 transmembrane domain and a signaling domain connected to the intracellular domain of CD3 (e.g., CD3-ζ). In some embodiments, the intracellular signaling domain includes a chimeric CD28 and CD137 (4-1BB, TNFRSF9) co-stimulatory domain connected to the intracellular domain of CD3ζ.

[0491] In some embodiments, the receptor contains one or more, such as two or more, co-stimulatory domains and activation domains in the cytoplasmic portion, such as a major activation domain. Exemplary receptors contain intracellular components of CD3-ζ, CD28, and 4-1BB.

[0492] In some embodiments, the CAR or other antigen receptor, such as a TCR, also includes a marker, such as a cell surface marker, which can be used to confirm the transduction or engineering of cells to express the receptor, such as a truncated form of a cell surface receptor, such as a truncated EGFR (tEGFR). In some aspects, the marker comprises all or part (e.g., a truncated form) 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 operatively linked to a polynucleotide encoding a linker sequence (such as a cleavable linker sequence or a ribosomal skipping sequence, such as T2A). See WO2014031687. In some embodiments, the introduction of a construct encoding a CAR and EGFRt separated by a T2A ribosomal switch can express two proteins from the same construct, making EGFRt a marker for detecting cells expressing such a construct. In some embodiments, the marker and optional linker sequence can be any sequence disclosed in patent application publication WO2014031687. For example, the marker could be a truncated EGFR ((tEGFR)) that is optionally linked to a linker sequence, such as a T2A ribosomal skipping sequence.

[0493] In some implementations, the marker is a molecule, such as a cell surface protein, that is not naturally present on T cells or on T cells or parts thereof.

[0494] In some implementations, the molecule is a non-self molecule, such as a non-self protein, which is a molecule that the host's immune system does not recognize as "self" when the cell is adopted to it.

[0495] In some embodiments, the marker has no therapeutic function and / or does not produce an effect other than being used as a marker for genetic engineering (e.g., for selecting successfully engineered cells). In other embodiments, the marker may be a therapeutic molecule or a molecule that otherwise exerts certain desired effects, such as a ligand for cells encountered in vivo, such as a co-stimulatory or immune checkpoint molecule, thereby enhancing and / or attenuating the cellular response upon adoptive transfer and encounter with the ligand.

[0496] TCRs or CARs may contain one or more 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, leucine, α-aminodecanoic acid, homoserine, S-acetaminomethylcysteine, trans-3- and trans-4-hydroxyproline, 4-aminophenylalanine, 4-nitrophenylalanine, 4-chlorophenylalanine, 4-carboxyphenylalanine, 3-phenylserine, 3-hydroxyphenylalanine, phenylglycine, α-naphthylalanine, cyclohexylalanine, cyclohexylglycine, dihydroindole-2-carboxylic acid. Acids, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, aminomalonic acid, aminomalonic acid monoamide, N'-benzyl-N'-methyllysine, N,'N'-dibenzyllysine, 6-hydroxylysine, ornithine, α-aminocyclopentanecarboxylic acid, α-aminocyclohexanecarboxylic acid, α-aminocycloheptanecarboxylic acid, α-(2-amino-2-norbornene)carboxylic acid, α,γ-diaminobutyric acid, α,γ-diaminopropionic acid, homophenylalanine, and α-tert-butylglycine.

[0497] In some contexts, CARs are referred to as first-generation, second-generation, and / or third-generation CARs. In some respects, first-generation CARs are CARs that provide only CD3-chain-induced signaling after antigen binding; in some respects, second-generation CARs are CARs that provide both signaling and co-stimulatory signals, such as CARs containing intracellular signaling domains from co-stimulatory receptors (e.g., CD28 or CD137); and in some respects, third-generation CARs are CARs containing multiple co-stimulatory domains from different co-stimulatory receptors.

[0498] In some embodiments, the chimeric antigen receptor includes an extracellular portion containing an antibody or fragment described herein. In some aspects, the chimeric antigen receptor includes an extracellular portion and an intracellular signaling domain, wherein the extracellular portion contains an antibody or fragment described herein. In some embodiments, the antibody or fragment comprises an scFv or a single-domain VH antibody, and the intracellular domain contains ITAM. In some aspects, the intracellular signaling domain comprises the signaling domain of the ζ chain of CD3, i.e., the ζ(CD3) chain. In some embodiments, the chimeric antigen receptor includes a transmembrane domain connecting the extracellular domain and the intracellular signaling domain.

[0499] In some embodiments, the transmembrane domain contains the transmembrane portion of CD28. The extracellular domain and the transmembrane domain may be directly or indirectly connected. In some embodiments, the extracellular domain and the transmembrane domain are connected by a spacer, such as any spacer described herein. In some embodiments, the chimeric antigen receptor contains an intracellular domain of a T-cell costimulatory molecule, such as an intracellular domain between the transmembrane domain and the intracellular signaling domain. In some embodiments, the T-cell costimulatory molecule is CD28 or 41BB.

[0500] In some embodiments, the CAR contains an antibody (e.g., an antibody fragment), a transmembrane domain containing a transmembrane portion of CD28 or a functional variant thereof, and an intracellular signaling domain containing a signaling portion of CD28 or a functional variant thereof and a signaling portion of CD3ζ or a functional variant thereof. In some embodiments, the CAR contains an antibody (e.g., an antibody fragment), a transmembrane domain containing a transmembrane portion of CD28 or a functional variant thereof, and an intracellular signaling domain containing a signaling portion of 4-1BB or a functional variant thereof and a signaling portion of CD3ζ or a functional variant thereof. In some of these embodiments, the receptor also includes a spacer containing a portion of an Ig molecule (such as a human Ig molecule), such as an Ig hinge, for example an IgG4 hinge, or a hinge-only spacer.

[0501] In some implementations, the transmembrane domain of the receptor (e.g., CAR) is the transmembrane domain of human CD28 or a variant thereof, such as the 27-amino acid-sized transmembrane domain of human CD28 (accession number: P10747.1).

[0502] In some implementations, the chimeric antigen receptor contains an intracellular domain of a T-cell co-stimulatory molecule. In some aspects, the T-cell co-stimulatory molecule is CD28 or 41BB.

[0503] In some embodiments, the intracellular signaling domain includes an intracellular costimulatory signaling domain of human CD28 or a functional variant thereof or a portion thereof, such as its 41-amino acid-sized domain and / or a domain having LL to GG substitutions at positions 186-187 of the native CD28 protein. In some embodiments, the intracellular domain includes a 41BB intracellular costimulatory signaling domain or a functional variant thereof or a portion thereof, such as the 42-amino acid-sized cytoplasmic domain of human 4-1BB (accession number: Q07011.1) or a functional variant thereof or a portion thereof.

[0504] In some embodiments, the intracellular signal transduction domain includes the human CD3ζ stimulation signal transduction domain or a functional variant thereof, such as the 112AA cytoplasmic domain of isotype 3 of human CD3ζ (accession number: P20963.2), or the CD3ζ signal transduction domain described in U.S. Patent No. 7,446,190 or U.S. Patent No. 8,911,993.

[0505] In some embodiments, the spacer contains only the hinge region of IgG, such as a hinge of only IgG4 or IgG1. In other embodiments, the spacer is an Ig hinge attached to the CH2 and / or CH3 domains, such as an IgG4 hinge. In some embodiments, the spacer is an Ig hinge attached to both CH2 and CH3 domains, such as an IgG4 hinge. In some embodiments, the spacer is an Ig hinge attached only to the CH3 domain, such as an IgG4 hinge. In some embodiments, the spacer is or contains a glycine-serine-rich sequence or other flexible linker, such as known flexible linkers.

[0506] For example, in some embodiments, the CAR comprises an antibody or a fragment thereof, such as any HLA-peptide antibody comprising a single-chain antibody (sdAb, e.g., containing only the VH region) and the scFv described herein; a spacer, such as any Ig-hinge containing a spacer; a CD28 transmembrane domain; a CD28 intracellular signaling domain; and a CD3ζ signaling domain. In some embodiments, the CAR comprises an antibody or a fragment thereof, such as any HLA-peptide antibody comprising an sdAb and the scFv described herein; a spacer, such as any Ig-hinge containing a spacer; a CD28 transmembrane domain; a CD28 intracellular signaling domain; and a CD3ζ signaling domain.

[0507] Target-specific TCRs against A*01:01_ASSLPTTMNY (SEQ ID NO:260)[G10]

[0508] In some respects, this article provides an ABP comprising a TCR or an antigen-binding fragment thereof that specifically binds to an HLA-peptide target, wherein the HLA class I molecule of the HLA-peptide target is HLA subtype A*01:01, and the HLA-restricted peptide of the HLA-peptide target comprises the sequence ASSLPTTMNY (“G10”) (SEQ ID NO:260).

[0509] TCRs specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain an αCDR3 sequence. The αCDR3 sequence may be any of the αCDR3 sequences listed in Table 15. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety. Table 15 shows the α and βCDR3 sequences of identified TCR clonal types.

[0510] A TCR specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain a βCDR3 sequence. The βCDR3 sequence may be any of the βCDR3 sequences in Table 15. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety.

[0511] A TCR specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain a specific αCDR3 sequence and a specific βCDR3 sequence. For example, a TCR specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain an αCDR3 sequence and a βCDR3 sequence from any of the TCRs identified in Table 15. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety.

[0512] A TCR specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain TRAV, TRAJ, TRBV, optional TRBD and TRBJ amino acid sequences, optional TRAC sequence, and optional TRBC sequence. For example, a TCR specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain TRAV, TRAJ, TRBV, TRBD, TRBJ amino acid sequences, TRAC sequence, and TRBC sequence from any of the TCRs identified in Table 14. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety. For clarity, each identified TCR is assigned a TCR ID number. For example, TCR ID#1 assigned to TCR includes the TRAV25 sequence, TRAJ37 sequence, TRAC sequence, TRBV19 sequence, TRBD1 sequence, TRBJ1-5 sequence, and TRBC1 sequence.

[0513] A TCR specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain an αVJ sequence. The αVJ sequence may be any of the αVJ sequences listed in Table 16. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety.

[0514] A TCR specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain a βV(D)J sequence. The βV(D)J sequence may be any of the βV(D)J sequences in Table 16. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety.

[0515] A TCR specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain an αVJ sequence and a βV(D)J sequence. For example, a TCR specific to A*01:01_ASSLPTTMNY (SEQ ID NO:260) may contain an αVJ sequence and a βV(D)J sequence from any of the TCRs identified in Table 16. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety. Table 16 shows the full-length αV(J) and βV(D)J sequences of identified TCR clonal types. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety.

[0516] Target-specific TCR for A*01:01_HSEVGLPVY (SEQ ID NO:367)

[0517] In some respects, this article provides an ABP comprising a TCR or an antigen-binding fragment thereof that specifically binds to an HLA-peptide target, wherein the HLA class I molecule of the HLA-peptide target is HLA subtype A*01:01, and the HLA-restricted peptide of the HLA-peptide target comprises the sequence HSEVGLPVY (SEQ ID NO:367).

[0518] TCRs specific to A*01:01_HSEVGLPVY (SEQ ID NO:367) may contain an αCDR3 sequence. The αCDR3 sequence may be any of the αCDR3 sequences listed in Table 18. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety. Table 18 shows the α and βCDR3 sequences of identified TCR clonal types.

[0519] A TCR specific to A*01:01_HSEVGLPVY (SEQ ID NO:367) may contain a βCDR3 sequence. The βCDR3 sequence may be any of the βCDR3 sequences in Table 18. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety.

[0520] A TCR specific to A*01:01_HSEVGLPVY (SEQ ID NO:367) may contain a specific αCDR3 sequence and a specific βCDR3 sequence. For example, a TCR specific to A*01:01_HSEVGLPVY (SEQ ID NO:367) may contain an αCDR3 sequence and a βCDR3 sequence from any of the TCRs identified in Table 18. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety.

[0521] A TCR specific to A*01:01_HSEVGLPVY (SEQ ID NO:367) may contain TRAV, TRAJ, TRBV, optional TRBD and TRBJ amino acid sequences, optional TRAC sequence, and optional TRBC sequence. For example, a TCR specific to A*01:01_HSEVGLPVY (SEQ ID NO:367) may contain TRAV, TRAJ, TRBV, TRBD, TRBJ amino acid sequences, TRAC sequence, and TRBC sequence from any TCR identified in Table 17. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety.

[0522] A TCR specific to A*01:01_HSEVGLPVY (SEQ ID NO:367) may contain an αVJ sequence. The αVJ sequence may be any of the αVJ sequences listed in Table 19. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety.

[0523] A TCR specific to A*01:01_HSEVGLPVY (SEQ ID NO:367) may contain a βV(D)J sequence. The βV(D)J sequence may be any of the βV(D)J sequences in Table 19. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety.

[0524] A TCR specific to A*01:01_HSEVGLPVY (SEQ ID NO:367) may contain an αVJ sequence and a βV(D)J sequence. For example, a TCR specific to A*01:01_HSEVGLPVY (SEQ ID NO:367) may contain an αVJ sequence and a βV(D)J sequence from any of the TCRs identified in Table 19. Reference is made to PCT / US2018 / 06793, filed December 28, 2018, which is incorporated herein by reference in its entirety.

[0525] Target-specific TCRs targeting A*02:01__LLASSILCA (SEQ ID NO:322)[G7]

[0526] In some respects, this article provides an ABP comprising a TCR or an antigen-binding fragment thereof that specifically binds to an HLA-peptide target, wherein the HLA class I molecule of the HLA-peptide target is HLA subtype A*02:01, and the HLA-restricted peptide of the HLA-peptide target comprises the sequence LLASSILCA (SEQ ID NO:322).

[0527] TCRs specific to A*02:01__LLASSILCA (SEQ ID NO:322) may contain the αCDR3 sequence. Reference is made to SEQ ID NOs 4277, 4278, 4279, 4280, or 4281 of PCT / US2018 / 046997, filed August 17, 2018, which are incorporated herein by reference in their entirety.

[0528] TCRs specific to A*02:01__LLASSILCA (SEQ ID NO:322) may contain the βCDR3 sequence. Reference is made to SEQ ID NOs 4291-4295 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0529] A TCR specific to A*02:01__LLASSILCA (SEQ ID NO:322) may contain a specific αCDR3 sequence and a specific βCDR3 sequence. For a specific combination of αCDR3 and βCDR3 sequences, refer to PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0530] A TCR specific to A*02:01__LLASSILCA (SEQ ID NO:322) may comprise TRAV, TRAJ, TRBV, optional TRBD and TRBJ amino acid sequences, optional TRAC sequence, and optional TRBC sequence. For specific combinations of TRAV, TRAJ, TRBV, optional TRBD, TRBJ amino acid sequences, optional TRAC sequence, and optional TRBC sequence, refer to PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0531] TCRs specific to A*02:01__LLASSILCA (SEQ ID NO:322) may contain the αVJ sequence. Reference is made to SEQ ID NOs4306-4310 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0532] TCRs specific to A*02:01__LLASSILCA (SEQ ID NO:322) may contain the βV(D)J sequence. Reference is made to SEQ ID NOs 4321-4325 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0533] The TCR specific to A*02:01__LLASSILCA (SEQ ID NO:322) may contain αVJ and βV(D)J sequences. For specific combinations of αVJ and βV(D)J sequences, refer to PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0534] Target-specific TCR for A*01:01_EVDPIGHLY (SEQ ID NO:354)

[0535] In some respects, this article provides an ABP comprising a TCR or an antigen-binding fragment thereof that specifically binds to an HLA-peptide target, wherein the HLA class I molecule of the HLA-peptide target is HLA subtype A*01:01, and the HLA-restricted peptide of the HLA-peptide target comprises the sequence EVDPIGHLY (SEQ ID NO:354).

[0536] TCRs specific to A*01:01_EVDPIGHLY (SEQ ID NO:354) may contain the αCDR3 sequence. Reference is made to SEQ ID NOS 3052-3350 or 4273-4276 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0537] TCRs specific to A*01:01_EVDPIGHLY (SEQ ID NO:354) may contain the βCDR3 sequence. Reference is made to SEQ ID NOs3351-3655 or 4287-4290 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0538] A TCR specific to A*01:01_EVDPIGHLY (SEQ ID NO:354) may contain a specific αCDR3 sequence and a specific βCDR3 sequence. For a specific combination of αCDR3 and βCDR3 sequences, refer to PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0539] A TCR specific to A*01:01_EVDPIGHLY (SEQ ID NO:354) may comprise TRAV, TRAJ, TRBV, optional TRBD and TRBJ amino acid sequences, optional TRAC sequence, and optional TRBC sequence. For specific combinations of TRAV, TRAJ, TRBV, optional TRBD, TRBJ amino acid sequences, optional TRAC sequence, and optional TRBC sequence, refer to PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0540] TCRs specific to A*01:01_EVDPIGHLY (SEQ ID NO:354) may contain the αVJ sequence. Reference is made to SEQ ID NOs3656-3961 or 4302-4305 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0541] TCRs specific to A*01:01_EVDPIGHLY (SEQ ID NO:354) may contain the βV(D)J sequence. Reference is made to SEQ ID NOS 3962-4269 or 4317-4320 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0542] A TCR specific to A*01:01_EVDPIGHLY (SEQ ID NO:354) may contain an αVJ sequence and a βV(D)J sequence. For specific combinations of αVJ and βV(D)J sequences, refer to PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0543] Target-specific TCRs against B*44:02_GEMSSNSTAL (SEQ ID NO:357)

[0544] In some respects, this article provides an ABP comprising a TCR or an antigen-binding fragment thereof that specifically binds to an HLA-peptide target, wherein the HLA class I molecule of the HLA-peptide target is HLA subtype B*44:02, and the HLA-restricted peptide of the HLA-peptide target comprises the sequence GEMSSNSTAL (SEQ ID NO:357).

[0545] TCRs specific to B*44:02_GEMSSNSTAL (SEQ ID NO:357) may contain the αCDR3 sequence. Reference is made to SEQ ID NOs 4284-4286 or 3138 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0546] TCRs specific to B*44:02_GEMSSNSTAL (SEQ ID NO:357) may contain the βCDR3 sequence. Reference is made to SEQ ID NOS 4298-4301 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0547] A TCR specific to B*44:02_GEMSSNSTAL (SEQ ID NO:357) may contain a specific αCDR3 sequence and a specific βCDR3 sequence. For specific combinations of αCDR3 and βCDR3 sequences, refer to PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0548] A TCR specific to B*44:02_GEMSSNSTAL (SEQ ID NO:357) may comprise TRAV, TRAJ, TRBV, optional TRBD and TRBJ amino acid sequences, optional TRAC sequence, and optional TRBC sequence. For specific combinations of TRAV, TRAJ, TRBV, optional TRBD, TRBJ amino acid sequences, optional TRAC sequence, and optional TRBC sequence, refer to PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0549] TCRs specific to B*44:02_GEMSSNSTAL (SEQ ID NO:357) may contain the αVJ sequence. Reference is made to SEQ ID NOs4313-4316 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0550] TCRs specific to B*44:02_GEMSSNSTAL (SEQ ID NO:357) may contain the βV(D)J sequence. Reference is made to SEQ ID NOS 4328-4331 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0551] A TCR specific to B*44:02_GEMSSNSTAL (SEQ ID NO:357) may contain an αVJ sequence and a βV(D)J sequence. For specific combinations of αVJ and βV(D)J sequences, refer to PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0552] Target-specific TCRs against A*02:01_GVYDGEEHSV (SEQ ID NO:356)

[0553] In some respects, this article provides an ABP comprising a TCR or an antigen-binding fragment thereof that specifically binds to an HLA-peptide target, wherein the HLA class I molecule of the HLA-peptide target is HLA subtype A*02:01, and the HLA-restricted peptide of the HLA-peptide target comprises the sequence GVYDGEEHSV (SEQ ID NO:356).

[0554] TCRs specific to A*02:01_GVYDGEEHSV (SEQ ID NO:356) may contain the αCDR3 sequence. Reference is made to SEQ ID NOs 4282-4283 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0555] TCRs specific to A*02:01_GVYDGEEHSV (SEQ ID NO:356) may contain the βCDR3 sequence. Reference is made to SEQ ID NOs 4296-4297 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0556] A TCR specific to A*02:01_GVYDGEEHSV (SEQ ID NO:356) may contain a specific αCDR3 sequence and a specific βCDR3 sequence. For specific combinations of αCDR3 and βCDR3 sequences, refer to PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0557] A TCR specific to A*02:01_GVYDGEEHSV (SEQ ID NO:356) may comprise TRAV, TRAJ, TRBV, optional TRBD and TRBJ amino acid sequences, optional TRAC sequence, and optional TRBC sequence. For specific combinations of TRAV, TRAJ, TRBV, optional TRBD, TRBJ amino acid sequences, optional TRAC sequence, and optional TRBC sequence, refer to PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0558] TCRs specific to A*02:01_GVYDGEEHSV (SEQ ID NO:356) may contain the αVJ sequence. Reference is made to SEQ ID NOS 4311-4312 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0559] TCRs specific to A*02:01_GVYDGEEHSV (SEQ ID NO:356) may contain the βV(D)J sequence. Reference is made to SEQ ID NOs 4326-4327 of PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0560] A TCR specific to A*02:01_GVYDGEEHSV (SEQ ID NO:356) may contain an αVJ sequence and a βV(D)J sequence. For specific combinations of αVJ and βV(D)J sequences, refer to PCT / US2018 / 046997, filed August 17, 2018, which is incorporated herein by reference in its entirety.

[0561] engineered cells

[0562] Cells, such as cells containing antigen receptors, are also provided, for example, antigen receptors containing the extracellular domain of the anti-HLA-peptide ABP as described herein (e.g., CAR or TCR). Populations of such cells and compositions containing such cells are also provided. In some embodiments, the composition or population is rich in such cells, such as cells expressing the HLA-peptide ABP, comprising at least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or greater than 99% of all cells in the composition, or a certain type of cell (e.g., T cells or CD8+ or CD4+ cells). In some embodiments, the composition comprises at least one cell containing an antigen receptor disclosed herein. The composition includes pharmaceutical compositions and formulations for administration (e.g., for adoptive cell therapy). Treatment methods for administering the cells and compositions to a subject, such as a patient, are also provided.

[0563] Therefore, genetically engineered cells expressing ABP containing a receptor (e.g., TCR or CAR) are also provided. These cells are generally eukaryotic cells, such as mammalian cells, and are typically human cells. In some embodiments, the cells are derived from blood, bone marrow, lymph nodes, or lymphoid organs, and are cells of the immune system, such as cells of innate or adaptive immunity, such as myeloid or lymphoid cells containing lymphocytes, typically T cells and / or NK cells. Other exemplary cells include stem cells, such as pluripotent and multipotent stem cells, including induced pluripotent stem cells (iPSCs). The cells are generally primary cells, such as cells isolated directly from a subject and / or cells isolated from and frozen from a subject. In some implementations, the cells comprise one or more subgroups of T cells or other cell types, such as the entire T cell population, CD4+ cells, CD8+ cells, and their subpopulations, as defined by function, activation state, maturity, differentiation potential, expansion, recycling, localization and / or persistence, antigen specificity, type of antigen receptor, presence in a specific organ or compartment, marker or cytokine secretion profile, and / or degree of differentiation. Regarding the subject to be treated, the cells may be allogeneic and / or autologous. These methods include off-the-shelf approaches. In some aspects, as with the prior art, the cells are pluripotent and / or multipotent, such as stem cells, such as induced pluripotent stem cells (iPSCs). In some implementations, the method includes isolating cells from the subject as described herein, preparing, processing, culturing, and / or engineering them, and re-introducing them into the same patient before or after cryopreservation.

[0564] Subtypes and subsets of T cells and / or CD4+ and / or CD8+ T cells include: primary T (TN) cells, effector T cells (TEFF), memory T cells and their subtypes, such as stem cell memory T (TSCM) cells, central memory T (TCM) cells, effector memory T (TEM) cells or terminally differentiated effector memory T cells, tumor-infiltrating lymphocytes (TIL), immature T cells, mature T cells, helper T cells, cytotoxic T cells, mucosa-associated non-variant T (MALT) cells, naturally occurring and adopted regulatory T (Treg) cells, helper T cells, such as TH1 cells, TH2 cells, TH3 cells, TH17 cells, TH9 cells, TH22 cells, follicular helper T cells, α / β T cells, and δ / γ T cells.

[0565] In some embodiments, the cells are natural killer (NK) cells. In some embodiments, the cells are monocytes or granulocytes, such as bone marrow cells, macrophages, neutrophils, dendritic cells, mast cells, eosinophils, and / or basophils.

[0566] Cells can be genetically modified to reduce or knock out endogenous TCR expression. Such modifications are described in the following literature: Mol Ther Nucleic Acids. Dec 2012; 1(12):e63; Blood. Aug 11, 2011; 118(6):1495-503; Blood. Jun 14, 2012; 119(24):5697–5705; Torikai, Hiroki et al. "HLA and TCR Knockout by Zinc Finger Nucleases: Toward “off-the-Shelf” Allogeneic T-Cell Therapy for CD19+ Malignancies." Blood 116.21(2010):3766;Blood.January 18, 2018;131(3):311-322.doi:10.1182 / blood-2017-05-787598;and WO2016069283, which are incorporated in whole by reference.

[0567] Cells can be genetically modified to promote the secretion of cytokines. This modification is described by Hsu C, Hughes MS, Zheng Z, Bray RB, Rosenberg SA, Morgan RA. Primary human T lymphocytes engineered with a codon-optimized IL-15gene resist withdrawal-induced apoptosis and persist long-term in the absence of exogenous cytokine. J Immunol. 2005; 175:7226–34; Quintarelli C, Vera JF, Savoldo B, GiordanoAttianese GM, Pule M, Foster AE, Co-expression of cytokine and suicide genes to enhance the activity and safety of tumor-specific cytotoxic Tlymphocytes. Blood. 2007;110:2793–802; and Hsu C, Jones SA, Cohen CJ, Zheng Z, Kerstann K, Zhou J, Cytokine-independent growth and clonal expansion of aprimary human CD8+T-cell clone following retroviral transduction with the IL-15gene. Blood. 2007;109:5168–77.

[0568] It has been shown that the mismatch between chemokine receptors on T cells and chemokines secreted by tumors is the cause of suboptimal transport of T cells into the tumor microenvironment. To improve therapeutic efficacy, cells can be genetically modified to increase their recognition of chemokines in the tumor microenvironment. Examples of such modifications are described in Moon et al., Expression of afunctional CCR2 receptor enhances tumor localization and tumor eradication by targeted human T cells expressing a mesothelin-specific chimeric antibody receptor. Clin Cancer Res. 2011; 17:4719-4730(()); and Craddock et al., Enhanced tumor trafficking of GD2 chimeric antigen receptor T cells by expression of the chemokine receptor CCR2b. J Immunother. 2010; 33:780-788.

[0569] Cells can be genetically modified to enhance the expression of costimulatory / enhancing receptors (such as CD28 and 41BB).

[0570] Adverse reactions to T-cell therapy can include cytokine release syndrome and persistent B-cell exhaustion. Introducing a suicide / safety switch into recipient cells can improve the safety of cell therapy. Therefore, cells can be genetically modified to include a suicide / safety switch. A suicide / safety switch can be a gene that confers sensitivity to a reagent (e.g., a drug) on cells expressing the gene and causes cell death when the cell comes into contact with or is exposed to the reagent. An exemplary suicide / safety switch is described in Protein Cell. 2017 Aug; 8(8):573–589. The suicide / safety switch can be HSV-TK. The suicide / safety switch can be a cytosine deaminase, a purine nucleoside phosphorylase, or a nitroreductase. The suicide / safety switch can be RapaCIDe. TM The suicide / safety switch system can be CD20 / rituximab, as described in U.S. Patent Application Publication No. US20170166877A1. The suicide / safety switch system can be CD20 / rituximab, as described in Haematologica. 2009 Sep; 94(9):1316–1320. These references are incorporated herein by reference in their entirety.

[0571] TCRs or CARs can be introduced into recipient cells as cleavage receptors, which assemble only in the presence of heterodimerized small molecules. Such systems are described in Science. 2015. 16 October; 350(6258):aab4077 and U.S. Patent No. 9,587,020 (the entire documents and patents are hereby incorporated by reference).

[0572] In some embodiments, the cell contains one or more nucleic acids, such as polynucleotides encoding the TCRs or CARs disclosed herein, wherein said polynucleotides are introduced through genetic engineering to express the recombinant TCRs or CARs disclosed herein or genetically engineered TCRs or CARs. In some embodiments, the nucleic acid is heterologous, i.e., not typically present in the cell or in samples obtained from the cell, such as samples obtained from another organism or cell, for example, cells that are not typically found in engineered cells and / or organisms from which such cells are derived. In some embodiments, the nucleic acid is not naturally occurring, such as a nucleic acid not found in nature, comprising a chimeric combination of nucleic acids encoding various domains from multiple different cell types.

[0573] Nucleic acids can contain codon-optimized nucleotide sequences. Without being bound by any specific theory or mechanism, codon optimization of nucleotide sequences is believed to increase the translation efficiency of mRNA transcripts. Nucleotide sequence codon optimization can involve replacing one codon encoding the same amino acid with a natural codon, but it can improve translation efficiency by translating tRNA, which is more readily available within the cell. Nucleotide sequence optimization can also reduce mRNA secondary structures that interfere with translation, thereby improving translation efficiency.

[0574] TCRs or CARs can be introduced into recipient cells using constructs or vectors. Exemplary constructs are described herein. The polynucleotides encoding the α and β chains of a TCR or CAR may be present in a single construct or in separate constructs. The polynucleotides encoding the α and β chains are operatively linked to a promoter, such as a heterologous promoter. The heterologous promoter can be a strong promoter, such as EF1α, CMV, PGK1, Ubc, β-actin, CAG, etc. The heterologous promoter can be a weak promoter. The heterologous promoter can be an inducible promoter. Exemplary inducible promoters include, but are not limited to, TRE, NFAT, GAL4, LAC, etc. Other exemplary inducible expression systems are described in U.S. Patent Nos. 5,514,578, 6,245,531, 7,091,038 and European Patent No. 0517805, which are incorporated herein by reference in their entirety.

[0575] Constructs for introducing TCRs or CARs into recipient cells may also contain polynucleotides encoding signal peptides (signal peptide elements). Signal peptides can facilitate surface transport of the introduced TCR or CAR. Exemplary signal peptides include, but are not limited to, CD8 signal peptides, immunoglobulin signal peptides, with specific examples including GM-CSF and IgGκ. Such signal peptides are described in Trends Biochem Sci. Oct 2006; 31(10):563-71. Epub Aug 21, 2006; and An et al., “Construction of a New Anti-CD19 Chimeric Antigen Receptor and the Anti-Leukemia Function Study of the Transduced T Cells.” Oncotarget 7.9(2016):10638–10649. PMC.Web. Aug 16, 2018; which are hereby incorporated by reference.

[0576] In some cases, such as when both the α and β chains are expressed from a single construct or open reading frame, or when the construct contains a marker gene, the construct may contain a ribosomal jumping sequence. The ribosomal jumping sequence may be a 2A peptide, such as a P2A or T2A peptide. Exemplary P2A and T2A peptides are described in Scientific Reports, Volume 7, Article No. 2193(2017)(())(()), which is hereby incorporated in its entirety by reference. In some cases, a FURIN / PACE cleavage site is introduced upstream of the 2A element. For example, the FURIN / PACE cleavage site is described at http: / / www.nuolan.net / substrates.html. The cleavage peptide may also be a cleavage site for factor Xa. In cases where both the α and β chains are expressed from a single construct or open reading frame, the construct may contain an internal ribosomal entry site ((IRES)).

[0577] The construct may further comprise one or more marker genes. Exemplary marker genes include, but are not limited to, GFP, luciferase, HA, and lacZ. As known to those skilled in the art, the marker may be a selective marker, such as an antibiotic resistance marker, a heavy metal resistance marker, or a biocidal resistance marker. The marker may be a complementary marker for auxotrophic hosts. Exemplary complementary markers and auxotrophic hosts are described in Gene. 2001 Jan 24; 263(1-2):159-69. Such markers may be expressed via IRES, frameshift sequences, 2A peptide linkers, fusions with TCRs or CARs, or separately from a single promoter.

[0578] Exemplary vectors or systems for introducing TCR or CAR into recipient cells include, but are not limited to: adeno-associated virus, adenovirus, adenovirus + modified vaccinia virus, ankara virus (MVA), adenovirus + retrovirus, adenovirus + Sendai virus, adenovirus + vaccinia virus, alphavirus (VEE) replicon vaccine, antisense oligonucleotide, Bifidobacterium longum, CRISPR-Cas9, Escherichia coli, flavivirus, gene gun, herpesvirus, mononucleotic herpesvirus, Lactococcus lactis, electroporation, lentivirus, liposome transfection, Listeria monocytogenes, measles virus, modified vaccinia ankara virus (MVA), mRNA electroporation, naked / plasmid DNA, naked / plasmid DNA + adenovirus, naked / plasmid DNA + modified vaccinia ankara virus (MVA), naked / plasmid DNA + RNA transfer, naked / plasmid DNA + vaccinia virus, naked / plasmid DNA + vesicular stomatitis virus, Newcastle disease virus, non-viral vectors, PiggyBac TM (PB) transposons, nanoparticle-based systems, poliovirus, poxvirus, poxvirus + vaccinia virus, retrovirus, RNA transfer, RNA transfer + naked / plasmid DNA, RNA virus, Saccharomyces cerevisiae, Salmonella typhimurium, Semliki Forest virus, Sendai virus, Shigella dysenteriae, simian virus, siRNA, Sleeping Beauty transposon, Streptococcus mutans, vaccinia virus, Venezuelan equine encephalitis virus replicon, vesicular stomatitis virus, and Vibrio cholerae.

[0579] In a preferred embodiment, TCR or CAR is transmitted via adeno-associated virus (AAV), adenovirus, CRISPR-CAS9, herpesvirus, lentivirus, liposome transfection, mRNA electroporation, or PiggyBac. TM (PB) transposons, retroviruses, RNA transfer, or Sleeping Beauty transposons are introduced into recipient cells.

[0580] In some implementations, the vector used to introduce TCR or CAR into recipient cells is a viral vector. Exemplary viral vectors include adenovirus vectors, adeno-associated virus (AAV) vectors, lentiviral vectors, herpesvirus vectors, retroviral vectors, etc. Such vectors are described herein.

[0581] Figure 2Exemplary embodiments of a TCR construct for introducing a TCR or CAR into recipient cells are shown. In some embodiments, the TCR construct comprises the following polynucleotide sequences in the 5'-3' direction: a promoter sequence, a signal peptide sequence, a TCRβ variable (TCRβv) sequence, a TCRβ constant (TCRβc) sequence, a cleavage peptide (e.g., P2A), a signal peptide sequence, a TCRα variable (TCRαv) sequence, and a TCRα constant (TCRαc) sequence. In some embodiments, the TCRβc and TCRαc sequences of the construct comprise one or more murine regions, such as the complete murine constant sequence or human-to-mouse amino acid exchange described herein. In some embodiments, the construct further comprises the 3' of the TCRαc sequence, a cleavage peptide sequence (e.g., T2A), and a reporter gene. In one embodiment, the construct comprises the following polynucleotide sequences in the 5'-3' direction: a promoter sequence, a signal peptide sequence, a TCRβ variable ((TCRβv)) sequence, a TCRβ constant (TCRβc) sequence containing one or more murine regions, a cleavage peptide (e.g., P2A), a signal peptide sequence, a TCRα variable (TCRαv) sequence, and a TCRα constant (TCRαc) sequence, wherein the TCRα constant (TCRαc) sequence contains one or more murine regions, a cleavage peptide (e.g., T2A), and a reporter gene.

[0582] Figure 3 An exemplary construct backbone sequence for cloning TCRs into an expression system for therapeutic development is described.

[0583] Figure 4 An exemplary construct sequence is described for cloning an identified A*0201-LLASSILCA (SEQ ID NO:322)-specific TCR into an expression system for therapeutic development.

[0584] Figure 5 An exemplary construct sequence is described for cloning an identified A*0101_EVDPIGHLY (SEQ ID NO:354) specific TCR into an expression system for therapeutic development.

[0585] Nucleotides, vectors, host cells, and related methods

[0586] Also provided are isolated nucleic acids encoding the HLA-peptide ABP, a vector containing the nucleic acid, a host cell containing the vector and the nucleic acid, and a recombinant technology for producing the ABP.

[0587] Nucleic acids can be recombinant nucleic acids. Recombinant nucleic acids can be constructed outside living cells by linking natural or synthetic nucleic acid fragments to nucleic acid molecules or their replication products that can replicate in living cells. For the purposes of this article, replication can be in vitro or in vivo.

[0588] For recombinant production of ABP, the nucleic acid encoding ABP can be isolated and inserted into a reproducible vector for further cloning (i.e., DNA amplification) or expression. In some respects, the nucleic acid can be produced by homologous recombination, for example, as described in U.S. Patent No. 5,204,244, which is incorporated in whole by reference.

[0589] Many different vectors are known in the art. 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, for example, as described in U.S. Patent No. 5,534,615, which is incorporated herein by reference in its entirety.

[0590] Exemplary vectors or constructs suitable for expressing ABPs (e.g., TCRs, CARs, antibodies, or their antigen-binding fragments) include, for example, the pUC series (Fermentas Life Sciences), the pBluescript series (Stratagene, La Jolla, CA), the pET series (Novagen, Madison, WI), the pGEX series (Pharmacia Biotech, Uppsala, Sweden), and the pEX series (Clontech, Palo Alto, CA). Phage vectors such as AGT10, AGT11, AZapII (Stratagene), AEMBL4, and ANM1149 are also suitable for expressing the ABPs described herein.

[0591] Illustrative examples of suitable host cells are provided below. These host cells are not restrictive, and any suitable host cell can be used to generate the ABP provided herein.

[0592] Suitable host cells include any prokaryotic (e.g., bacteria), lower eukaryotic (e.g., yeast), or higher eukaryotic (e.g., mammalian) cells. Suitable prokaryotes include eubacteria, such as Gram-negative or Gram-positive organisms, such as Enterobacteriaceae, including species such as Escherichia (E. coli), Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella, Salmonella typhimurium, and Serratia (Serratia muscaria). *S. marcescans*, *Shigella*, *Bacilli* (*B. subtilis* and *B. licheniformis*), *Pseudomonas* (*P. aeruginosa*), and *Streptomyces*. A useful Escherichia coli cloning host is *Escherichia coli* 294, although other strains such as *Escherichia coli* B, *Escherichia coli* X1776, and *Escherichia coli* W3110 are also applicable.

[0593] Besides prokaryotes, eukaryotic microorganisms (such as filamentous fungi or yeasts) are also suitable clonal or expression hosts for vectors encoding the HLA-peptide ABP. *Saccharomyces cerevisiae* or common baker's yeast is a commonly used lower eukaryotic host microorganism. However, many other genera, species, and strains are available and useful, such as *Schizosaccharomyces*; *Kluyveromyces* (including *Kluyveromyces lactis*, *Kluyveromyces brittle*, *Kluyveromyces bulgaricus*, *Kluyveromyces wickii*, *Kluyveromyces flavum*, *Kluyveromyces davidiana*, *Kluyveromyces thermophilus*, and *Kluyveromyces marx*); *Yersinia*; *Pichia pastoris*; *Candida* (*Candida albicans*); *Trichoderma*; *Neurospora crassa*; *Schwanyophytes* (*Schwanyophytes serrata*); and filamentous fungi, such as *Penicillium*, *Cereus*, and *Aspergillus* (*Aspergillus nidus* and *Aspergillus niger*).

[0594] Useful mammalian host cells include COS-7 cells, HEK293 cells; young hamster kidney (BHK) cells; Chinese hamster ovary (CHO) cells; mouse testicular supporting cells; and African green monkey kidney cells (VERO-76), etc.

[0595] Host cells used to produce the HLA-peptide ABP can be cultured in a variety of media. Commercially available media, such as Ham F10, Minimal Essential Medium (MEM), RPMI-1640, and DuPont's modified Eagle's Minimal Essential Medium (DMEM), are suitable for culturing host cells. Additionally, any media described in Ham et al., Meth. Enz., 1979, 58:44; Barnes et al., Anal. Biochem., 1980, 102:255; and U.S. Patents 4,767,704, 4,657,866, 4,927,762, 4,560,655, and 5,122,469; or WO 90 / 03430 and WO 87 / 00195, all of which are incorporated herein by reference in their entirety.

[0596] Any of these culture media may 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 (defined as inorganic compounds typically present in micromolar final concentrations), and glucose or equivalent energy. Other necessary supplements may also be included at appropriate concentrations known to those skilled in the art.

[0597] The culture conditions, such as temperature and pH, are the same as those previously used with the host cells used for expression, and are obvious to the average technician.

[0598] When using recombinant techniques, ABP can be generated intracellularly, in the periplasmic space, or secreted directly into the culture medium. If ABP is generated intracellularly, the first step is to remove particle debris from the host cell or cell lysate, for example, by centrifugation or ultrafiltration. For example, Carter et al. (Bio / Technology, 1992, 10:163-167, incorporated herein by reference in its entirety) described a method for isolating ABP secreted into the periplasmic space of Escherichia coli. Briefly, the cell paste is thawed for approximately 30 minutes in the presence of sodium acetate (pH 3.5), EDTA, and benzyl sulfonyl fluoride (PMSF). Cell debris can then be removed by centrifugation.

[0599] In some implementations, ABP is produced in a cell-free system. In some aspects, the cell-free system is an in vitro transcription and translation system, as described in Yin et al. mAbs, 2012, 4:217-225, which is incorporated herein by reference in its entirety. In some aspects, the cell-free system utilizes a cell-free extract from eukaryotic or prokaryotic cells. In some aspects, the prokaryotic cell is *Escherichia coli*. Cell-free expression of ABP may be useful, for example, when ABP accumulates in cells as insoluble aggregates or when the yield obtained from periplasmic expression is low.

[0600] When ABP is secreted into the culture medium, a commercially available protein concentrate filter (e.g., [filter name missing]) is generally used first. or The ultrafiltration unit concentrates the supernatant from this type of expression system. Protease inhibitors such as PMSF may be included in any of the above steps to inhibit proteolysis, and antibiotics may be included to prevent the growth of foreign contaminants.

[0601] Cell-prepared ABP compositions can be purified using, for example, hydroxyapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography, with affinity chromatography being a particularly useful purification technique. The suitability of protein A as an affinity ligand depends on the type and isotype of any immunoglobulin Fc domain present in the ABP. Protein A can be used to purify ABP containing human γ1, γ2, or γ4 heavy chains (Lindmark et al., J. Immunol. Meth., 1983, 62:1-13, which are incorporated herein by reference in their entirety). Protein G can be used for all mouse isotypes and human γ3 (Guss et al., EMBO J., 1986, 5:1567-1575, which are incorporated herein by reference in their entirety).

[0602] The matrix to which the affinity ligands are attached is typically agarose, but other matrices can also be used. Mechanically stable matrices (such as controlled-pore glass or poly(divinyl)styrene) offer faster flow rates and shorter processing times than agarose. If the ABP contains a CH3 domain, it can be used... The resin is purified.

[0603] Other protein purification techniques may also be used by those skilled in the art, such as ion exchange column fractionation, ethanol precipitation, reversed-phase high-performance liquid chromatography (HPLC), silica gel chromatography, and heparin. Chromatography, focusing chromatography, SDS-PAGE, and ammonium sulfate precipitation are among the methods used.

[0604] After any initial purification step, the mixture containing the target ABP and contaminants can be subjected to low-pH hydrophobic interaction chromatography using an elution buffer with a pH between about 2.5 and about 4.5, typically at low salt concentrations (e.g., about 0 to about 0.25 M salt).

[0605] Methods for preparing HLA-peptide ABP

[0606] Preparation of HLA-peptide antigen

[0607] The HLA-peptide antigen used to isolate or generate the ABP described herein can be a complete HLA-peptide or a fragment of an HLA-peptide. The HLA-peptide antigen can be, for example, an isolated protein or a protein expressed on the cell surface.

[0608] In some implementations, the HLA-peptide antigen is a non-natural variant of the HLA-peptide, such as an HLA-peptide protein with an amino acid sequence not found in nature or a post-translational modified HLA-peptide protein.

[0609] In some embodiments, the HLA-peptide antigen is truncated by removing, for example, intracellular or transmembrane sequences or signaling sequences. In some embodiments, the HLA-peptide antigen is fused to a human IgG1 Fc domain or a multihistidine tag at its C-terminus.

[0610] Methods for identifying ABP

[0611] ABPs that bind to HLA-peptides can be identified using any method known in the art, such as phage display or subject immunization.

[0612] One method for identifying antigen-binding proteins includes: providing at least one HLA-peptide target; binding said at least one target to the antigen-binding protein, thereby identifying said antigen-binding protein. Antigen-binding proteins can be present in a library containing a variety of different antigen-binding proteins.

[0613] In some implementations, the library is a phage display library. Phage display libraries can be developed such that they are substantially free of HLA antigen-binding proteins that nonspecifically bind to HLA-peptide targets. Antigen-binding proteins can be present in yeast display libraries containing a variety of different antigen-binding proteins. Yeast display libraries can be developed such that they are substantially free of HLA antigen-binding proteins that nonspecifically bind to HLA-peptide targets.

[0614] In some implementations, the library is a yeast display library.

[0615] In some implementations, the library is a TCR presentation library. Exemplary TCR presentation libraries and their usage are described in: WO 98 / 39482; WO 01 / 62908; WO 2004 / 044004; WO2005116646, WO2014018863, WO2015136072 and WO2017046198; and Helmut et al. ((2000)) PNAS 97(26)14578-14583, all of which are incorporated herein by reference in their entirety.

[0616] In some respects, the combination step is performed more than once, optionally at least three times, for example at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 times.

[0617] Additionally, the method may further 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.

[0618] Another method for identifying antigen-binding proteins 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. Isolation of the antigen-binding protein may include: screening the serum of the subject to identify the antigen-binding protein. The method may further 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.

[0619] In some aspects, isolating antigen-binding proteins includes isolating B cells from a subject expressing the antigen-binding protein. The B cells can be used to generate hybridomas. The B cells can also be used to clone one or more CDRs of B cells. For example, the B cells can be immortalized by EBV conversion. The sequence encoding the antigen-binding protein can be cloned from immortalized B cells or can be cloned directly from B cells isolated from an immunized subject. A library containing the antigen-binding protein from the B cells can also be created, optionally, wherein the library is a phage display library or a yeast display library.

[0620] Another method for identifying antigen-binding proteins may include: obtaining cells containing the antigen-binding protein; contacting the cells with an HLA-multimer (e.g., a tetramer) containing at least one HLA-peptide target; and identifying the antigen-binding protein by the binding between the HLA-multimer and the antigen-binding protein.

[0621] The cells may be, for example, T cells, optionally cytotoxic T lymphocytes (CTLs) or natural killer (NK) cells. The method may further include isolating cells optionally using flow cytometry, magnetic separation, or single-cell separation. The method may further include sequencing the antigen-b...

Claims

1. An isolated antigen-binding protein (ABP) that specifically binds to a human leukocyte antigen (HLA)-peptide target, wherein the HLA-peptide target comprises 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 is an HLA subtype A*02:01, and the HLA-restricted peptide comprises the sequence AIFPGAVPAA, and wherein the ABP comprises an antibody or an antigen-binding fragment thereof, the antibody or antigen-binding fragment thereof comprising CDR-H1 composed of the sequence GTLSSYPIN, CDR-H2 composed of the sequence GWISTYSGHADYA, CDR-H3 composed of the sequence CARSYDYGDYLNFDYW, CDR-L1 composed of the sequence QASQDISNYLN, CDR-L2 composed of the sequence AASSLQS, and CDR-L3 composed of the sequence CQQSYSIPPTF.

2. The isolated ABP as claimed in claim 1, wherein the ABP comprises the VH sequence QVQLVQSGAEVKKPGASVKVSCKASGGTLSSYPINWVRQAPGQGLEWMGWISTYSGHADYAQKLQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARSYDYGDYLNFDYWGQGTLVTVSS.

3. The isolated ABP as described in claim 1 or 2, wherein the ABP comprises the VL sequence DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSIPPTFGGGTKVDIK.

4. The detachable ABP as claimed in claim 1, wherein the ABP is connected to the support.

5. The isolated ABP as claimed in claim 4, wherein the scaffold comprises serum albumin or Fc.

6. The isolated ABP as described in claim 5, wherein the Fc is a human Fc and is an isotype of IgG, IgA, IgD, IgE or IgM.

7. The detachable ABP as claimed in claim 1, wherein the ABP is connected to the support via a connector.

8. The isolated ABP as claimed in claim 7, wherein the adapter is a peptide adapter.

9. The isolated ABP as claimed in claim 8, wherein the peptide linker is the hinge region of a human antibody.

10. The isolated ABP as claimed in claim 1, wherein the ABP comprises an Fv fragment, a Fab fragment, an F(ab')2 fragment, a Fab' fragment, a scFv fragment, a scFv-Fc fragment, and / or a single-domain antibody or an antigen-binding fragment thereof.

11. The isolated ABP as claimed in claim 1, wherein the ABP comprises an scFv fragment.

12. The isolated ABP as claimed in claim 1, wherein the ABP is a monoclonal antibody.

13. The isolated ABP of claim 1, wherein the ABP is an antibody, and wherein the antibody is a humanized antibody, a human antibody, or a chimeric antibody.

14. The isolated ABP as claimed in claim 1, wherein the ABP is multispecific.

15. The isolated ABP as claimed in claim 14, wherein the ABP is bispecific.

16. The isolated ABP of claim 1, wherein the ABP binds to more than one antigen or more than one epitope on a single antigen.

17. The isolated ABP of claim 1, wherein the ABP comprises a heavy chain constant region selected from the classes of IgG, IgA, IgD, IgE and IgM.

18. The isolated ABP of claim 1, wherein the ABP comprises a heavy chain constant region of class human IgG and subclasses selected from IgG1, IgG4, IgG2 and IgG3.

19. The isolated ABP of claim 1, wherein the ABP comprises a modification to extend its half-life.

20. The isolated ABP of claim 1, wherein the ABP comprises a decorated Fc.

21. The isolated ABP of claim 20, wherein the modified Fc comprises one or more mutations that extend the half-life.

22. The isolated ABP of claim 21, wherein one or more mutations that extend the half-life are YTE.

23. The isolated ABP of claim 1, wherein the ABP is part of a chimeric antigen receptor (CAR), the chimeric antigen receptor comprising: an extracellular portion comprising the ABP; and an intracellular signal transduction domain.

24. The isolated ABP of claim 23, wherein the ABP comprises scFv and the intracellular signal transduction domain comprises ITAM.

25. The isolated ABP as claimed in claim 23 or 24, wherein the intracellular signal transduction domain comprises the signal transduction domain of the ζ chain of the CD3-ζ (CD3) chain.

26. The isolated ABP of claim 23, further comprising a transmembrane domain connecting the extracellular portion and the intracellular signal transduction domain.

27. The isolated ABP of claim 26, wherein the transmembrane domain comprises the transmembrane portion of CD28.

28. The isolated ABP as described in claim 23, further comprising an intracellular signal transduction domain of a T cell co-stimulatory molecule.

29. The isolated ABP as claimed in claim 28, wherein the T cell co-stimulatory molecule is CD28, 4-1BB, OX-40, ICOS, or any combination thereof.

30. The isolated ABP of claim 1, wherein the ABP binds to the HLA-peptide target via a contact point with the HLA class I molecule and via a contact point with the HLA-restricted peptide of the HLA-peptide target.

31. The isolated ABP of claim 1, wherein the binding of the ABP to the amino acid position or contact point on the restriction peptide or HLA subtype is determined by position scanning, hydrogen-deuterium exchange, or protein crystallography.

32. An engineered cell expressing a receptor comprising the isolated ABP of any one of claims 1-31.

33. The engineered cell as described in claim 32 is a T cell.

34. The engineered cell of claim 33, wherein the T cell is a cytotoxic T cell (CTL).

35. The engineered cell of claim 32 or 33, wherein the ABP is expressed from a heterologous promoter.

36. An isolated polynucleotide or polynucleotide group encoding the isolated ABP, or its antigen-binding portion, as claimed in any one of claims 1-31.

37. A vector or group of vectors comprising the polynucleotide or group of polynucleotides as described in claim 36.

38. A host cell comprising the polynucleotide or polynucleotide group as described in claim 36, or the vector or vector group as described in claim 37.

39. The host cell of claim 38, wherein the host cell is a CHO, HEK293, or a T cell.

40. A method for generating ABP, comprising: The ABP is expressed using the host cell as described in claim 38, and the expressed ABP is isolated.

41. A pharmaceutical composition comprising isolated ABP as described in any one of claims 1-31, and a pharmaceutically acceptable excipient.

42. A kit comprising isolated ABP as claimed in any one of claims 1-31, or a pharmaceutical composition as claimed in claim 41, and instructions for use.

43. A virus comprising the isolated polynucleotides or polynucleotide groups as described in claim 36.

44. The virus of claim 43, wherein the virus is a filamentous bacteriophage.

45. A yeast cell comprising the isolated polynucleotide or polynucleotide group as described in claim 36.

Citation Information

Patent Citations

Methods and materials for high gradient magnetic separation of biological materials
EP0452342A1
Trifunctional antibody-like compounds as a combined diagnostic and therapeutic agent
EP0453082A1
Identification and expression of insect steroid receptor DNA sequences
EP0517805A1
Constitutive expression of costimulatory ligands on adoptively transferred T lymphocytes
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Halogenated pyranthrone and process of making same
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Patent Information

AI Technical Summary

Abstract

Description

Patent Citations

Methods and materials for high gradient magnetic separation of biological materials

Trifunctional antibody-like compounds as a combined diagnostic and therapeutic agent

Identification and expression of insect steroid receptor DNA sequences

Constitutive expression of costimulatory ligands on adoptively transferred T lymphocytes

Halogenated pyranthrone and process of making same