TCR molecules and cells targeting the KRAS G12V mutation and uses thereof

Abstract

The present invention provides a binding protein having antigen specificity for an antigenic peptide / HLA complex, comprising a binding domain containing a T cell receptor (TCR) α chain variable region and a TCR β chain variable region, and uses of the binding protein and pharmaceutical compositions containing the binding protein in the treatment of cancers associated with the antigenic peptide.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority to a Chinese patent application bearing application number 202310507375.6, filed with the China Patent Office on May 06, 2023, the entire contents of which are hereby incorporated by reference.

[0002] This specification relates to TCR molecules and cells expressing the TCR molecules, particularly TCR molecules that target target cells with a KRAS G12V mutation and immune effector cells expressing the TCR molecules. This specification also relates to the use of these TCR molecules and cells in therapy. [Background technology]

[0003] The RAS gene family (HRAS, KRAS, NRAS) is the most common proto-oncogene leading to tumorigenesis. KRAS mutations (centred at codon 12) are the most common and are present in approximately 22-30% of tumors, particularly in pancreatic adenocarcinoma (~60-90%), colorectal cancer (~40%), lung cancer (~10% in China, ~30% in Europe and the United States), bile duct cancer (~17%-37%), ovarian cancer, and endometrial cancer (~10-40%, depending on pathological classification, often occurring in serous carcinoma). Taking pancreatic adenocarcinoma as an example, approximately 60,430 patients are expected to be diagnosed with the disease in 2021, and approximately 48,220 patients are expected to die from the disease. While progress has been made in diagnosis and treatment of pancreatic adenocarcinoma over the past decade, the lack of effective treatment options means the 5-year survival rate is approximately 9%. Currently, treatment for non-resectable pancreatic adenocarcinoma primarily involves chemotherapy to shrink tumors, but this is limited by the emergence of drug resistance, and targeted therapy has also had limited success. Furthermore, pancreatic adenocarcinoma is also resistant to immunotherapy, possibly due in part to a lack of new antigen-reactive tumor-infiltrating lymphocytes due to the disease's low mutational burden. Research into KRAS-G12V-targeting TCR-T may potentially alleviate the current predicament of a lack of effective treatments for KRAS-mutated tumors. Summary of the Invention

[0004] In one aspect, the present disclosure provides a binding protein having antigenic peptide / HLA complex antigen specificity, the binding protein comprising a binding domain containing a T cell receptor (TCR) alpha chain variable region and a TCR beta chain variable region, the antigenic peptide comprises the amino acid sequence VVGAVGVGK (SEQ ID NO: 91); the HLA is HLA-A*11:01, the CDR3 of the α chain variable region comprises an amino acid sequence set forth in SEQ ID NO: 5, 15, 25, 35, 45, 55, 65, 75, or 85, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity to the amino acid sequence set forth in SEQ ID NO: 5, 15, 25, 35, 45, 55, 65, 75, or 85; The CDR3 of the β-chain variable region comprises an amino acid sequence set forth in SEQ ID NO: 10, 20, 30, 40, 50, 60, 70, 80, or 90, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity to the amino acid sequence set forth in SEQ ID NO: 10, 20, 30, 40, 50, 60, 70, 80, or 90.

[0005] In some embodiments, the antigenic peptide is VVGAVGVGK (SEQ ID NO: 91) or VVVGAVGVGK (SEQ ID NO: 92).

[0006] In some embodiments, the α chain variable region has: 1) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 3, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 4, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 5; 2) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 13, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 14, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 15 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 15; or 3) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 23, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 24, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 25 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 25; 4) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 33, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 34, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 35 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 35; 5) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 43, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 44, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 45 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 45; 6) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 53, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 54, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 55 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 55; 7)CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 63, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 64, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 65 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 65; 8) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 73, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 74, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 75 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 75; or 9) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 83, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 84, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 85 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 85; In the β-chain variable region, 1) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 8, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 9, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 10 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 10; 2) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 18, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 19, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 20 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 20; 3) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 28, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 29, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 30 or an amino acid sequence set forth in SEQ ID NO: 30, 4) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 38, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 39, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 40, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 40; 5) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 48, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 49, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 50, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 50; 6) 6) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 58, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 59, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 60, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 60;CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 68, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 69, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 70 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 70; 8) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 78, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 79, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 80 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 80; or 9) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 88, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 89, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 90 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 90, includes amino acid sequences having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence shown.

[0007] In some embodiments, 1) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO:3, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:4, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:5 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:5; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO:8, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:9, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:10 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:10; or 2) 2) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 13, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 14, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 15 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 15; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 18, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 19, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 20 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 20; 3) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 23, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 24, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 25 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 25; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 28, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 29, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 30 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 30;4) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 33, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 34, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 35 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 35; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 38, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 39, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 40 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 40; or 5) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 43, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 44, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 45, 6) an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 45, wherein in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 48, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 49, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 50 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 50; In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 53, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 54, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 55 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 55; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 58, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 59, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 60 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 60; or 7)In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 63, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 64, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 65 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 65; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 68, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 69, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 70 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 70; or 8) In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 73, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 74, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 75 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 75; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 78, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 79, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 80 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 80; or In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 83, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 84, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 85 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 85; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 88, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 89, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 90 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 90.

[0008] In another aspect, the description provides a binding protein comprising a binding domain containing a T cell receptor (TCR) alpha chain variable region and a TCR beta chain variable region, wherein: 1) in the alpha chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO:3, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:4, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:5 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:5; and in the beta chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO:8, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:9, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:10 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:10; or 2) 2) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 13, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 14, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 15 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 15; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 18, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 19, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 20 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 20;3) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 23, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 24, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 25 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 25; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 28, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 29, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 30 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 30; 4) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 33, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 34, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 35 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 35; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 38, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 39, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 40 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 40; or 5) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 43, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 44, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 45, 6) an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 45, wherein in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 48, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 49, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 50 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 50;In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 53, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 54, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 55 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 55; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 58, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 59, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 60 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 60; or 7) In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 63, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 64, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 65 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 65; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 68, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 69, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 70 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 70; or 8) In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 73, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 74, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 75 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 75; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 78, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 79, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 80 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 80; orIn the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 83, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 84, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 85 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 85; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 88, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 89, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 90 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 90.

[0009] In some embodiments, the binding protein comprises: 1) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO:2 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity to the amino acid sequence set forth in SEQ ID NO:2; and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO:7 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity to the amino acid sequence set forth in SEQ ID NO:7; or 2) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 12 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 12, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 17 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 17; 3) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 22 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 22, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 27 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 27; 4) 4) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 32 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 32, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 37 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 37;the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 42 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 42, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 47 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 47; 6) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 52 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 52, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 57 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 57; 7) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 62 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 62, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 67 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 67; 8) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 72 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 72, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 77 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 77; or 9)The α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 82 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 82, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 87 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 87.

[0010] In some embodiments, the binding protein comprises the TCR alpha chain and TCR beta, wherein 1) the alpha chain comprises the amino acid sequence set forth in SEQ ID NO: 1 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity to the amino acid sequence set forth in SEQ ID NO: 1, and the beta chain comprises the amino acid sequence set forth in SEQ ID NO: 6 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity to the amino acid sequence set forth in SEQ ID NO: 6; or 2) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 11 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 11, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 16 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 16; 3) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 21 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 21, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 26 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 26; 4) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 31 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 31; the β chain comprises the amino acid sequence set forth in SEQ ID NO: 36 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 36; or the α chain comprises the amino acid sequence set forth in SEQ ID NO: 41;41, wherein the β chain comprises the amino acid sequence set forth in SEQ ID NO: 46 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 46; 6) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 51 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 51; the β chain comprises the amino acid sequence set forth in SEQ ID NO: 56 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 56; 6) the α chain comprises an amino acid sequence set forth in SEQ ID NO: 71 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 71; and the β chain comprises an amino acid sequence set forth in SEQ ID NO: 76 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 76; or 9)The α chain comprises the amino acid sequence set forth in SEQ ID NO: 81 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 81, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 86 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 86.

[0011] In some embodiments, the binding protein is a protein that is: 1) TCR molecule; 2) an antibody molecule; or 3) CAR molecule.

[0012] In some embodiments, when T cells expressing the binding protein are incubated with HLA-A*11:01 cells in the presence of the antigenic peptide, or when T cells expressing the binding protein are incubated with HLA-A*11:01 cells expressing the antigenic peptide, the T cells specifically kill the HLA-A*11:01 cells.

[0013] In some embodiments, when T cells expressing the binding protein are incubated with HLA-A*11:01 cells in the presence of the antigenic peptide, or when T cells expressing the binding protein are incubated with HLA-A*11:01 cells expressing the antigenic peptide, the T cells produce INF-γ and / or express CD69, or the amount of INF-γ and / or CD69 expression is increased.

[0014] In some embodiments, the binding protein further comprises a binding entity covalently or non-covalently linked to the binding domain, preferably a binding entity that may be a detectable marker, a radioisotope, or a therapeutic agent.

[0015] In another aspect, the document provides an isolated nucleic acid molecule encoding the binding protein, an alpha chain variable region, a beta chain variable region, an alpha chain, or a beta chain.

[0016] In another aspect, the present disclosure provides a vector comprising the above-described nucleic acid molecule.

[0017] In another aspect, the present disclosure provides a host cell that expresses the binding protein or that expresses the nucleic acid molecule or vector.

[0018] In some embodiments, the host cell is a mammalian cell, preferably a human cell.

[0019] In some embodiments, the host cell is a T cell or an NK cell.

[0020] In some embodiments, the host cells have killing activity against HLA-A*11:01 cells expressing the antigenic peptide VVGAVGVGK (SEQ ID NO: 91) or VVVGAVGVGK (SEQ ID NO: 92).

[0021] In another aspect, the present disclosure provides a pharmaceutical composition comprising 1) the binding protein, nucleic acid molecule, vector, or host cell described above, and 2) a pharmaceutically acceptable carrier.

[0022] In another aspect, the description provides the use of the binding protein, nucleic acid molecule, vector or host cell described above in the manufacture of a medicament for treating a tumor expressing the antigenic peptide.

[0023] In another aspect, the description provides a method of treating a tumor in a subject, comprising administering to the subject a therapeutically effective amount of the binding protein, nucleic acid molecule, vector, host cell, or pharmaceutical composition described above.

[0024] In some embodiments, the tumor is selected from pancreatic adenocarcinoma, colorectal cancer, lung cancer, bile duct cancer, endometrial cancer, and ovarian cancer.

[0025] In another aspect, the present disclosure provides a detection kit comprising the above-described binding protein. [Brief explanation of the drawings]

[0026] [Figure 1] FIG. 1 shows representative flow cytometry graphs of sorting of HLA-A*11:01 / KRAS G12V-specific T cells by pentamer enrichment of stimulated T cells. [Figure 2A] Figure 2A shows the killing results of CD8+ T cells transduced with different HLA-A*11:01 / KRAS G12V-specific TCRs and untransduced CD8+ T cells against K562-A*11:01 target cells bearing different antigenic peptides. The polypeptides carried in the figure are G12V-9 peptide: VVGAVGVGK (SEQ ID NO: 91), G12V-10 peptide: VVVGAVGVGK (SEQ ID NO: 92), and WT peptide: VVGAGGVGK (SEQ ID NO: 93) + VVVGAGGVGK (SEQ ID NO: 94), respectively. [Figure 2B] Figure 2B shows the killing of CD8+ T cells transduced with different HLA-A*11:01 / KRAS G12V-specific TCRs and untransduced CD8+ T cells against BxPC3 cells overexpressing HLA-A*11:01 and / or KRAS G12V. BxPC3-A1101 are BxPC3 cells transduced with HLA-A*11:01, BxPC3-A1101-G12V are BxPC3 cells transduced simultaneously with HLA-A*11:01 and KRAS G12V, and BxPC3-A1101-G12D are BxPC3 cells transduced simultaneously with HLA-A*11:01 and KRAS G12D. [Figure 3]Figure 3 shows the release of IFN-γ after co-incubation of TCR-transduced T cells with target cells bearing the polypeptide. BxPC3-A1101 are BxPC3 cells transduced with HLA-A*11:01, BxPC3-G12V are BxPC3 cells transduced with KRAS G12V, and BxPC3-A1101-G12V are BxPC3 cells transduced simultaneously with HLA-A*11:01 and KRAS G12V. [Figure 4A] Figure 4A shows the release of IFN-γ detected by ELISPOT after co-incubation of PBMCs transduced with a given TCR with target cells (K562-A*11:01) bearing different concentrations of polypeptide (10-11-10-6 M). [Figure 4B] Figure 4B shows the CD69 expression profile detected by flow cytometry after co-incubation of Jurkat cells transduced with a given TCR with target cells (K562-A*11:01) bearing different concentrations of polypeptide (10-11-10-6 M). [Figure 4C] Figure 4C shows the CD69 expression profile detected by flow cytometry after co-incubation of Jurkat cells transduced with a given TCR with target cells (K562-A*11:01) carrying different concentrations of G12V-9 or G12V-10 polypeptide (10-11-10-6M). DETAILED DESCRIPTION OF THE INVENTION

[0027] Unless otherwise explained, all technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art.

[0028] The term "or," unless clearly stated otherwise in the context of the context, refers to one of listed alternatives. The term "and / or" refers to any one, any two, any three, any more, or all of the listed alternatives.

[0029] The term "about" generally refers to a variation of more or less than 10% of the specified numerical value, for example, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or a variation of more or less than 10% of the specified numerical value.

[0030] The term "comprise" or "include" means the inclusion of the aforementioned elements, integers, or steps, but does not exclude any other elements, integers, or steps. As used herein, unless otherwise specified, the terms "comprise" or "include" also include embodiments consisting of the aforementioned elements, integers, or steps. For example, when referring to a variable region of a specific sequence, this also includes a variable region consisting of that specific sequence.

[0031] The term "major histocompatibility complex (MHC)" refers to the set of genes encoding the major histocompatibility antigens in animals. The structures of MHC class I and MHC class II molecules have now been fully characterized, and they are members of the immunoglobulin superfamily. Human MHC is called HLA (human leukocyte antigen). HLA class I molecules primarily contain the protein products (heavy or α chains) of three functional genes: HLA-A, HLA-B, and HLA-C. Each class I HLA heavy chain is matched with β2-microglobulin (β2m) to form a complete MHC class I molecule in the form of a heterodimer. Due to the polymorphism of the genes encoding HLA molecules, the HLA Nomenclature Committee has specified naming principles for specific alleles, including the following: for a given allele, the three letters HLA are followed by the name of the locus of the gene, separated by a hyphen, e.g., HLA-A; the locus name is followed by an * symbol, followed by the numerical number of the gene family to which the allele belongs, preferably consistent with the corresponding serological classification, followed by a colon (:) and the specific numerical number of the allele, e.g., HLA-A*02:01. One of the main functions of MHC molecules is antigen presentation: they bind antigenic peptides through their peptide-binding groove to form antigenic peptide-MHC complexes (pMHC), which are then presented on the cell surface for recognition by T cells (via TCRs) to trigger an immune response.

[0032] The term "antigenic peptide" refers to a short peptide capable of binding to the peptide-binding groove of an MHC molecule (or HLA molecule). The antigen-binding groove of an HLA class I molecule is composed of the α1 and α2 domains of the heavy chain (the extracellular region of which contains three domains, α1, α2, and α3), each of which folds into an α-helix and four β-sheets. Two α-helices form the walls of the groove, and eight β-sheets form the bottom of the groove. The length of an antigenic peptide suitable for fitting into the binding groove is typically 8 to 11 amino acids. In one specific example, the antigenic peptide is a fragment of a mutated KRAS protein: VVGAVGVGK (SEQ ID NO: 91). In another specific example, the antigenic peptide is a fragment of a mutated KRAS protein: VVVGAVGVGK (SEQ ID NO: 92).

[0033] The term "antigenic peptide / MHC complex" or "antigenic peptide / HLA complex" refers to a complex (pMHC) composed of an MHC molecule and an antigenic peptide, where the antigenic peptide is located in the peptide-binding groove. A TCR molecule recognizes and specifically binds to the complex via the CDR sequences in its binding domain. Binding of the TCR molecule to the pMHC complex is MHC-dependent; i.e., the TCR molecule recognizes the pMHC complex while simultaneously recognizing the antigenic peptide.

[0034] The term "T cell receptor (TCR)" refers to a functional unit of T cells that recognizes antigens (pMHC) and belongs to the immunoglobulin superfamily. TCRs expressed by T cells are glycoproteins present on the surface of the cell membrane in the form of α / β or γ / δ chain heterodimers. In the human body, most TCRs in peripheral blood (90%-95%) are heterodimers composed of two polypeptide chains, α and β. Both the α and β chains are divided into variable regions (Vα, Vβ), constant regions (Cα, Cβ), transmembrane regions, and intracellular regions. The intracellular regions of both the α and β chains are short, consisting of only a few amino acids. The sequence variation of the constant regions (Cα, Cβ) is relatively small, and specific amino acid sequences can be found, for example, in the public database of the International Immunogenetics Information System (IMGT). The transmembrane region contains positively charged amino acids that noncovalently bind to negatively charged amino acids in the transmembrane region of the CD3 molecule, stabilizing the structure of the TCR-CD3 complex. Amino acid sequence analysis of the variable regions of TCRs reveals that Vα and Vβ each contain three hypervariable regions, called complementarity-determining regions (CDRs), namely, CDR1, CDR2, and CDR3. CDR3 is the most frequently mutated region and largely determines the antigen specificity of the TCR. Similar to antibody molecules, apart from the hypervariable regions, the variable regions also contain four framework regions (FR1-FR4). The locations of CDR1-CDR3 and FR1-FR4 in the full-length TCR sequence are defined by the IMGT nomenclature, which is well known and can be found in the IMGT public database. The variable regions in the α chain and β chain, particularly the three CDRs in the α chain and the three CDRs in the β chain, together form a portion of the TCR that specifically recognizes and binds to the pMHC complex (also referred to herein as the "binding domain"). In the immune system, binding of an antigen-specific TCR to a pMHC complex induces direct physical contact between the T cell and an antigen-presenting cell (APC) or target cell, thereby activating the T cell or achieving a killing effect on the target cell.Unless otherwise stated or implied in the context, when referring to a TCR, the term also includes functional fragments thereof having antigen specificity, for example, the variable regions of Vα and Vβ linked via a short peptide chain.

[0035] The term "antibody" is used herein in its broadest sense and refers to a protein or polypeptide, including immunoglobulins or other types of molecules, that contain one or more antigen-binding domains and exhibit binding specificity to a specific antigen. Specific examples of antibodies may include intact antibodies (e.g., a typical four-chain antibody molecule), single-chain antibodies, single-domain antibodies, multispecific antibodies, etc. A typical antibody molecule is usually a tetramer composed of two identical heavy chains and two identical light chains linked together via disulfide bonds. Conserved differences in amino acid sequence separate the heavy and light chains into a variable region (V) located at the amino terminus and a constant region (C) located at the carboxy terminus. The variable region is responsible for antigen recognition and binding, while the constant region (e.g., Fc fragment) is responsible for initiating downstream effects, such as antibody-dependent cell-mediated cytotoxicity (ADCC). The heavy and light chain variable regions each contain three local regions with a high degree of variation in amino acid composition and sequence order. These regions are important for antibody antigen binding and are therefore also called complementarity-determining regions (CDRs). The three CDRs of the heavy chain are called HCDR1, HCDR2, and HCDR3, respectively, and the three CDRs of the light chain are called LCDR1, LCDR2, and LCDR3, respectively. Each heavy chain variable region (VH) and light chain variable region (VL) consists of three CDRs and four FR regions, which may be arranged from the amino terminus to the carboxy terminus in the order FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. As used herein, the antibody molecule may also include a fusion protein reconstituted with the variable region of a TCR molecule and the constant region of an antibody molecule, which simultaneously possesses the antigen specificity of a TCR molecule and the effective function of the constant region of an antibody molecule.

[0036] The term "chimeric antigen receptor (CAR)" refers to an engineered membrane protein receptor molecule that can confer desired specificity, e.g., the ability to bind to a specific tumor antigen, to immune effector cells. A chimeric antigen receptor typically consists of an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. In some cases, the antigen-binding domain is a single scFv sequence or a single-domain antibody fragment, responsible for recognizing and binding to a specific antigen. The intracellular signaling domain typically contains an immunoreceptor tyrosine-based activation motif (ITAM), e.g., a signaling domain derived from a CD3ζ molecule, and is responsible for activating immune effector cells, resulting in their killing. A chimeric antigen receptor may also contain a signal peptide at the amino terminus responsible for intracellular localization of the nascent protein, and a hinge region between the antigen-binding domain and the transmembrane domain. In addition to the signaling domain, the intracellular signaling domain may further include a costimulatory domain, e.g., derived from a 4-1BB or CD28 molecule.

[0037] The terms "antigen specificity," "target," or "specific binding" refer to the ability of a TCR (or other fusion protein containing its binding domain, e.g., a CAR) to specifically recognize and bind with high affinity to a corresponding pMHC complex. Relative to other molecules present in the environment, a TCR (or other fusion protein containing its binding domain, e.g., a CAR) binds with higher affinity to another molecule (e.g., a pMHC complex) for which it has antigen specificity. Several parameters, e.g., EC 50 The binding affinity of TCR to pMHC complexes can be evaluated by measuring the KD value and KD value.

[0038] The term "binding protein" refers to a protein capable of specifically binding to a specific ligand (e.g., an antigen). As used herein, binding proteins may include TCR molecules (including single-chain TCRs and soluble TCR molecules) capable of specifically binding to a pMHC complex, fusion proteins consisting of the TCR molecule and other proteins (e.g., antibody molecules or chimeric antigen receptors), and immune complexes (linked with a detectable tag, therapeutic agent, or radioisotope, etc.).

[0039] The term "fusion protein" refers to a protein molecule that is artificially created (e.g., by genetic engineering) and is composed of at least two different peptide fragments that do not occur in nature or in a single protein molecule. Common examples of fusion proteins containing antibody fragments include multispecific antibodies, antibody-cytokine fusion proteins, enzyme-labeled antibodies for immunodetection, chimeric antigen receptors (CARs), etc.

[0040] As used herein, the term "variant" when referring to a TCR (or antigen-specific fragment thereof) refers to a protein obtained by introducing one or more amino acid insertions, deletions, or substitutions based on the parent TCR (or antigen-specific fragment thereof), while retaining at least some of the functions of the parent TCR (or antigen-specific fragment thereof) (particularly functions of interest, such as the ability to bind to a corresponding pMHC complex). For example, a variant of a TCR (or antigen-specific fragment thereof) retains at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the binding ability of the parent TCR (or antigen-specific fragment thereof) to a pMHC complex, or has a higher binding ability than the parent TCR (or antigen-specific fragment thereof). In some embodiments, a variant of a TCR (or antigen-specific fragment thereof) retains at least 80%, 85%, 90%, 95%, or even 100% or more of the binding affinity of the parent TCR (or antigen-specific fragment thereof) to an antigen. Variants of TCRs (or antigen-specific fragments thereof) typically include those in which amino acids are changed in the framework sequence of the variable region and / or the constant region, but alterations of one or a few amino acids in the CDR region sequence are not excluded. Therefore, those skilled in the art will understand that corresponding variants of TCRs (or antigen-specific fragments thereof) provided herein can be obtained by substituting, deleting, or adding a few amino acids based on the amino acid sequence (CDR sequence, variable region sequence, or α / β chain sequence) of a specific TCR (or antigen-specific fragment thereof) provided herein, and verifying or screening the binding ability or biological activity of the resulting product with the corresponding pMHC complex, and that these variants are also within the scope of the present invention.

[0041] The terms "nucleic acid molecule," "nucleic acid," and "polynucleotide" refer to a polymer of nucleotides and can be used interchangeably herein. Such nucleotide polymers may contain natural and / or non-natural nucleotides and include, but are not limited to, DNA, RNA, and PNA. A "nucleic acid sequence" refers to the linear sequence of nucleotides contained in a nucleic acid molecule or polynucleotide.

[0042] The term "vector" refers to a nucleic acid molecule (e.g., a nucleic acid plasmid, or a virus) that has been engineered to contain a polynucleotide of interest (e.g., a coding sequence for a polypeptide of interest) or that is capable of replicating in a host cell. A vector may contain one or more of the following elements: an origin of replication, regulatory sequences (e.g., a promoter and / or enhancer) that control the expression of one or more polynucleotides of interest, and / or one or more selectable marker genes (e.g., antibiotic resistance genes and genes that can be used in colorimetric assays, e.g., β-galactose). The term "expression vector" refers to a vector for expressing a polypeptide of interest in a host cell.

[0043] A "host cell" refers to a cell that can be or has been the recipient of a vector or an isolated polynucleotide. Host cells can be prokaryotic or eukaryotic. Exemplary eukaryotic cells include mammalian cells, e.g., primate or non-primate cells, fungal cells, e.g., yeast, plant cells, and insect cells. Non-limiting exemplary mammalian cells include CHO cells, HEK-293 cells, BHK cells, and PER-C6 cells, as well as cells derived therefrom, e.g., 293-6E, CHO-DG44, CHO-K1, CHO-S, and CHO-DS cells. In some embodiments, host cells are used to produce proteins of interest; for example, the TCR α and / or β chains provided herein can be secreted and produced by mammalian cells. In other embodiments, host cells possess a specific function after expressing a TCR molecule provided herein; for example, T cells possess the ability to recognize and kill target cells expressing a specific antigenic peptide after expressing a TCR molecule provided herein. A host cell may be an isolated cell or cell line, and includes cells transfected in vivo with a nucleic acid molecule or expression vector provided herein. A host cell includes the progeny of a single host cell, and the progeny may not necessarily be completely identical (in terms of morphology or genomic DNA complement) to the original parent cell due to natural, accidental, or deliberate mutation.

[0044] "Subject" includes animals, e.g., mammals, including, but not limited to, primates, rodents, simians, felines, canines, equines, bovines, porcines, sheep, goats, mammalian laboratory animals, mammalian farm animals, mammalian sport animals, and mammalian pets. A subject may be male or female and of any age, including infants, children, adolescents, adults, and geriatric subjects. In some instances, a subject refers to an individual in need of diagnosis or treatment for a disease or condition. In some instances, a subject receiving diagnosis or treatment may be a patient suffering from or at risk for the condition associated with the diagnosis or treatment. In certain instances, a subject is a human, e.g., a human patient. The term is generally used interchangeably with "patient," "subject to be tested," "subject to be treated," and the like.

[0045] When referring to a drug composition, the term "pharmaceutically acceptable carrier" refers to a solid or liquid diluent, filler, antioxidant, stabilizer, or other substance that can be used safely, and that is suitable for administration to humans and / or animals without undue adverse side effects, while at the same time maintaining the vitality of the drug or active agent therein.

[0046] When referring to the treatment of a disease, an "effective amount" refers to the amount of an active molecule (e.g., an antibody) sufficient to induce the biological or medical response in the subject's body that is desired by the clinician. The "effective amount" at the time of administration can be determined by one skilled in the art based on factors such as the route of administration, the subject's weight, age, and medical condition. For example, for protein-based drugs, a typical daily dosage range may be 0.01 mg-100 mg / kg body weight of the active ingredient. For immune effector cells (e.g., T cells) expressing the TCR provided herein, the number of cells per infusion administration may be, for example, about 1 x 10 6 to approximately 1 x 10 12 The number of cells may vary from 1 x 10 to 1 x 10 or more widely. 6Fewer than one T cell can be administered. Methods of administering the active molecules (e.g., TCRs, antibodies, fusion proteins) or immune effector cells provided herein include, but are not limited to, by injection, e.g., intravenously, intramuscularly, intraarterially, subcutaneously, intraperitoneally, etc.

[0047] When referring to amino acid or nucleotide sequences, the term "sequence identity" (also referred to as "sequence identity") refers to the amount of match between two amino acid or nucleotide sequences (e.g., a query sequence and a reference sequence), typically expressed as a percentage. Typically, before calculating the percentage of identity between two amino acid or nucleotide sequences, a sequence alignment is first performed to introduce gaps, if any. If the amino acid residues or bases in the two sequences are similar at a certain alignment position, the two sequences are considered to match at that position. If the amino acid residues or bases in the two sequences are different, the position is considered to be mismatched. Some algorithms calculate the sequence identity by dividing the number of matching positions by the total number of positions in the alignment window. Other algorithms further consider the number of gaps and / or gap length. For purposes of the present invention, the open alignment software BLAST (accessible at the homepage ncbi.nlm.nih.gov) can be used to obtain optimal sequence alignment with default settings and calculate the sequence identity between two amino acid or nucleotide sequences. In some embodiments, "at least 80% sequence identity" as used herein includes, but is not limited to, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or even 100% sequence identity.

[0048] The present specification provides an isolated or purified binding protein, e.g., a T cell receptor (TCR), antibody, or CAR, having antigen specificity for an antigenic peptide (having an amino acid mutation site relative to the wild-type sequence)-MHC complex, wherein the antigenic peptide is derived from a mutated KRAS protein.

[0049] Further provided herein are polypeptides and proteins related to the TCR or its binding domain, as well as related nucleic acid molecules, vectors (e.g., recombinant expression vectors), host cells (e.g., T cells), and pharmaceutical compositions.

[0050] Additionally, the present specification provides methods and kits for detecting the presence of a tumor in a subject, as well as methods for treating or preventing a tumor in a subject.

[0051] T cell receptor (TCR) The present specification provides an isolated or purified T cell receptor (TCR) having antigen specificity for an antigenic peptide (having an amino acid mutation site relative to the wild-type sequence)-MHC complex, wherein the antigenic peptide is derived from a mutated KRAS protein and is (1) VVGAVGVGK (SEQ ID NO: 91) or (2) VVVGAVGVGK (SEQ ID NO: 92), or consists thereof. Considering that MHC class I molecules do not strictly limit the length of antigenic peptides, the inventors predict that based on the above antigenic peptide sequences provided herein, the binding proteins or TCR molecules provided herein will also have the ability to bind to these antigenic peptide variant-MHC complexes even if one or two amino acids are added or deleted in the antigenic peptide (particularly at the terminus).

[0052] The TCRs provided herein may be αβ heterodimers or single-chain forms (scTCRs). Examples of single-chain forms include αβ TCR polypeptides in the form of Vα-L-Vβ, Vβ-L-Vα, Vα-Cα-L-Vβ, or Vα-L-Vβ-Cβ (where Vα and Vβ refer to the variable regions of the TCR α chain and TCR β chain, respectively; Cα and Cβ refer to the constant regions of the TCR α chain and TCR β chain, respectively; and L is a linker sequence, typically a short peptide). Linker sequences may be conventional in the art and are commonly used in antibody and TCR engineering. The TCRs provided herein may be water-soluble TCR molecules and do not contain the transmembrane regions of the α and β chains.

[0053] In some embodiments, the present specification provides a TCR having antigen specificity for the antigenic peptide VVGAVGVGK (SEQ ID NO: 91)-MHC complex or (2) VVVGAVGVGK (SEQ ID NO: 92)-MHC complex. The CDR3 of the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 5, 15, 25, 36, 45, 55, 65, 75, or 85, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 5, 15, 25, 36, 45, 55, 65, 75, or 85, and the CDR3 of the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 10, 20, 30, 40, 50, 60, 70, 80, or 90, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 10, 20, 30, 40, 50, 60, 70, 80, or 90. Preferably, the MHC molecule is HLA-A*11:01.

[0054] Furthermore, in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 3, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 3, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 5; or CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 13, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 14, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 15 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 15. or CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 23, CDR2 the amino acid sequence set forth in SEQ ID NO: 24, and CDR3 the amino acid sequence set forth in SEQ ID NO: 25, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 25; or CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 33, and CDR2 the amino acid sequence set forth in SEQ ID NO: 34; The amino acid sequence shown in sequence number 34, CDR3 comprises the amino acid sequence shown in SEQ ID NO: 35 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence shown in SEQ ID NO: 35, CDR1 comprises the amino acid sequence shown in SEQ ID NO: 43, CDR2 comprises the amino acid sequence shown in SEQ ID NO: 44, and CDR3 comprises the amino acid sequence shown in SEQ ID NO: 45 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% identity with the amino acid sequence shown in SEQ ID NO: 45. or an amino acid sequence having 95% identity, wherein CDR1 has the amino acid sequence set forth in SEQ ID NO: 53, CDR2 has the amino acid sequence set forth in SEQ ID NO: 54, and CDR3 has the amino acid sequence set forth in SEQ ID NO: 55, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 55, or wherein CDR1 has the amino acid sequence set forth in SEQ ID NO: 63, CDR2 has the amino acid sequence set forth in SEQ ID NO: 64, and CDR3 has the amino acid sequence set forth in SEQ ID NO: 65,or CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 73, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 74, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 75 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 75; or CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 83. the CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 84, the CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 85 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 85; and in the β chain variable region, the CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 8, the CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 9, and the CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 10 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 10. or CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 18, CDR2 the amino acid sequence set forth in SEQ ID NO: 19, and CDR3 the amino acid sequence set forth in SEQ ID NO: 20, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 20; or CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 28, CDR2 the amino acid sequence set forth in SEQ ID NO: 29, and CDR3 the amino acid sequence set forth in SEQ ID NO: the amino acid sequence set forth in SEQ ID NO: 30 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 30; CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 38; CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 39; and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 40 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 40;CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 48, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 49, CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 50 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 50, or CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 58, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 59, CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 60 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 60, or CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 68, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 69, CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 70 or an amino acid sequence set forth in SEQ ID NO: 7 or CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 78, CDR2 the amino acid sequence set forth in SEQ ID NO: 79, and CDR3 the amino acid sequence set forth in SEQ ID NO: 80 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 80; or CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 88, CDR2 the amino acid sequence set forth in SEQ ID NO: 89, and CDR3 the amino acid sequence set forth in SEQ ID NO: 90 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 90.

[0055] Furthermore, in the TCR molecule, in the α chain variable region, CDR1 comprises the amino acid sequence shown in SEQ ID NO: 3, CDR2 comprises the amino acid sequence shown in SEQ ID NO: 4, and CDR3 comprises the amino acid sequence shown in SEQ ID NO: 5 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence shown in SEQ ID NO: 5; and in the β chain variable region, CDR1 comprises the amino acid sequence shown in SEQ ID NO: 8, CDR2 comprises the amino acid sequence shown in SEQ ID NO: 9, and CDR3 comprises the amino acid sequence shown in SEQ ID NO: 10. or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence shown in SEQ ID NO: 10; or in the α chain variable region, CDR1 comprises the amino acid sequence shown in SEQ ID NO: 13, CDR2 comprises the amino acid sequence shown in SEQ ID NO: 14, and CDR3 comprises the amino acid sequence shown in SEQ ID NO: 15 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence shown in SEQ ID NO: 15; In the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 18, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 19, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 20 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 20; or in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 23, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 24, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 25 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 20. and wherein the β chain variable region comprises an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 25, wherein CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 28, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 29, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 30 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 30; and wherein the α chain variable region comprisesCDR1 comprises the amino acid sequence shown in SEQ ID NO: 33, CDR2 comprises the amino acid sequence shown in SEQ ID NO: 34, and CDR3 comprises the amino acid sequence shown in SEQ ID NO: 35 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence shown in SEQ ID NO: 35; and in the β chain variable region, CDR1 comprises the amino acid sequence shown in SEQ ID NO: 38, CDR2 comprises the amino acid sequence shown in SEQ ID NO: 39, and CDR3 comprises the amino acid sequence shown in SEQ ID NO: 40 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence shown in SEQ ID NO: 40. or in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 43, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 44, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 45 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 45; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 48. the α chain variable region comprises an amino acid sequence as set forth in SEQ ID NO: 53, CDR2 an amino acid sequence as set forth in SEQ ID NO: 54, and CDR3 an amino acid sequence as set forth in SEQ ID NO: 55 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence as set forth in SEQ ID NO: 55; and wherein in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 58, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 59, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 60 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 60; and in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 63,CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 64, CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 65 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 65, and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 68, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 69, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 70 or an amino acid sequence having at least 80% identity with the amino acid sequence set forth in SEQ ID NO: 70. % identity, preferably at least 85% identity, more preferably 90% or 95% identity, with the amino acid sequence set forth in SEQ ID NO: 75; or in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 73, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 74, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 75 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 75; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: or in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 83, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 84, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 85 or an amino acid sequence having at least 80%, preferably at least 85%, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 80; and the β chain variable region comprises an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 5, wherein CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 88, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 89, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 90 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 90.

[0056] In some specific embodiments, in the TCR molecule, the alpha chain comprises the amino acid sequence set forth in SEQ ID NO: 1 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 1; the beta chain comprises the amino acid sequence set forth in SEQ ID NO: 6 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 6; or the alpha chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 11. the β chain comprises the amino acid sequence set forth in SEQ ID NO: 16 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 16; the α chain comprises the amino acid sequence set forth in SEQ ID NO: 21 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 21. the β chain comprises the amino acid sequence set forth in SEQ ID NO:26 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:26; or the α chain comprises the amino acid sequence set forth in SEQ ID NO:31 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:31. the β chain comprises the amino acid sequence set forth in SEQ ID NO: 36 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 36; the α chain comprises the amino acid sequence set forth in SEQ ID NO: 41 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 41; the β chain comprises the amino acid sequence set forth in SEQ ID NO: 46;The α chain comprises an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 46, or the α chain comprises an amino acid sequence set forth in SEQ ID NO: 51 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 51, and the β chain comprises an amino acid sequence set forth in SEQ ID NO: 56 or an amino acid sequence having at least or the α chain comprises the amino acid sequence set forth in SEQ ID NO: 61 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 61, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 66 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 66. % identity, more preferably 90% or 95% identity, with the amino acid sequence set forth in SEQ ID NO: 71; or the α chain comprises an amino acid sequence set forth in SEQ ID NO: 71 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 71; and the β chain comprises an amino acid sequence set forth in SEQ ID NO: 76 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 76. or the α chain comprises the amino acid sequence set forth in SEQ ID NO: 81 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 81, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 86 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 86.

[0057] Those skilled in the art will understand that variants of the TCR molecules provided herein can be obtained by substituting, deleting, or adding a small number of amino acids based on the specific sequences provided herein, and verifying or screening the binding ability or biological activity of the resulting products with the corresponding pMHC complex, and these variants are also included within the scope of the present invention. For example, the TCR molecules provided herein may have at least one and no more than 10 amino acid modifications, for example, 5, 4, 3, 2, or 1 amino acid modifications, in their full length or variable region sequences or CDR sequences. For example, the TCR molecules may have at least one and fewer than 10 amino acid modifications, for example, 5, 4, 3, 2, or 1 amino acid modifications, in the α or β variable region sequences, and may have a total of fewer than 5, 4, 3, 2, or 1 amino acid modifications in the α or β variable region CDR sequences.

[0058] It is anticipated that the TCRs (or antigen-specific fragments thereof) described herein may contain conservative amino acid substitutions. Conservative amino acid substitutions are generally described as substitutions of one amino acid residue with another of similar chemical structure, while having little or no effect on the function, activity, or other biological properties of the polypeptide. Conservative amino acid substitutions are well known in the art. Conservative substitutions are, for example, substitutions of one amino acid from the following sets (a)-(e) for another amino acid from the same set: (a) small aliphatic nonpolar or weakly polar residues: Ala, Ser, Thr, Pro, and Gly; (b) polar negatively charged residues and their (uncharged) amides: Asp, Asn, Glu, and Gln; (c) polar positively charged residues: His, Arg, and Lys; (d) large aliphatic nonpolar residues: Met, Leu, Ile, Val, and Cys; and (e) aromatic residues: Phe, Tyr, and Trp.

[0059] In some embodiments, the TCRs (or antigen-specific fragments thereof) provided herein may further comprise post-translational modifications. Examples of post-translational protein modifications include phosphorylation, acetylation, methylation, ADP-ribosylation, ubiquitination, glycosylation, carbonylation, sumoylation, biotinylation, or the addition of polypeptide side chains or hydrophobic groups. Thus, the modified TCRs (or antigen-specific fragments thereof) may contain non-amino acid components, such as lipids, polysaccharides and monosaccharides, and phosphates. One form of glycosylation, for example, is sialylation modification, in which one or more sialic acid groups are attached to the polypeptide. Sialic acid groups improve the solubility and serum half-life of the protein, while also reducing the potential immunogenicity of the protein.

[0060] If the N-terminus of the amino acid sequence of a TCR provided herein is a methionine residue, said methionine may be removed during the production of the recombinant protein, as will be familiar to those skilled in the art.

[0061] Fusion proteins and immunoconjugates Additionally, the present specification provides fusion proteins comprising the variable regions of the α and / or β chains of the TCR molecules provided herein, in particular the CDR sequences thereof.

[0062] In some embodiments, the fusion protein is in the form of a single-chain TCR, for example, the α and β chain variable regions can be linked via a linker sequence to form a binding domain with antigen specificity for the complete corresponding pMHC complex.

[0063] In some embodiments, the variable regions of the α and / or β chains of the TCR molecules provided herein, particularly their CDR sequences, can be used to form multispecific TCR molecules. In some embodiments, the fusion protein comprises at least two functional moieties, a first functional moiety having antigen specificity for a first pMHC complex, and a second functional moiety having an antigen specificity similar to or different from the first functional moiety.

[0064] In some embodiments, the variable regions of the α and / or β chains of the TCR molecules provided herein, particularly their CDR sequences, can be used to form multispecific fusion proteins. In some embodiments, the fusion proteins comprise at least two functional moieties, where a first functional moiety has antigen specificity for a pMHC complex and a second functional moiety has antigen specificity or targeting different from that of the first functional moiety. In some embodiments, the second functional moiety can be a targeting moiety that can specifically recognize and bind to a tumor-specific antigen, tumor-associated antigen, or other target cell surface molecule, such as an antibody or antigen-binding fragment thereof (e.g., scFv), hormone, growth factor, cytokine, and any other natural or non-natural ligand that can bind to a cell surface receptor (e.g., epidermal growth factor receptor (EGFR), CD28, platelet-derived growth factor receptor (PDGFR), nicotinic acetylcholine receptor (nAChR), etc.).

[0065] In some embodiments, a TCR molecule (or α and β chain variable regions or a binding domain consisting of these) provided herein can be used to replace the binding domain of an antibody molecule (e.g., a typical tetrameric antibody molecule) to form a recombinant antibody molecule with the antigen specificity of the TCR molecule. In some embodiments, the α chain variable region of the TCR molecule replaces two heavy chain variable regions of the antibody molecule and is linked to the heavy chain constant region, while the β chain variable region replaces two light chain variable regions of the antibody molecule and is linked to the light chain constant region. In some embodiments, the β chain variable region of the TCR molecule replaces two heavy chain variable regions of the antibody molecule and is linked to the heavy chain constant region, while the α chain variable region replaces two light chain variable regions of the antibody molecule and is linked to the light chain constant region. In some embodiments, the α chain variable region and β chain variable region of the TCR molecule replace one heavy chain variable region and one light chain variable region of the antibody molecule, respectively, while maintaining one original antigen-binding domain of the antibody molecule, forming a recombinant antibody molecule with bispecificity that targets the pMHC complex and the target antigen of the original antibody molecule, respectively. The constant region (e.g., Fc fragment) of the recombinant antibody molecule thus formed can exert the functions of the antibody molecule, such as complement-dependent cytotoxicity (CDC), antibody-dependent cellular cytotoxicity (ADCC), and mediation of phagocytosis. Furthermore, fusion with an Fc fragment can increase the half-life of the TCR molecule (or the variable regions of the α and β chains or binding domains composed of these) in the body, so that the time between administrations of the TCR molecule (or the variable regions of the α and β chains or binding domains composed of these) can be increased when used as a therapeutic agent. In some embodiments, the Fc fragment may be derived from the constant region of an immunoglobulin, such as IgG1, IgG2, or IgG4.

[0066] In some embodiments, chimeric antigen receptors (CARs) can be constructed using the TCR molecules provided herein (or the variable regions of the α and β chains, or binding domains consisting of these) as the extracellular binding domain. For example, the variable regions of the α and β chains can be linked via a linker sequence to form a binding molecule similar to an scFv, and linked to a transmembrane region and an intracellular signaling domain instead of the extracellular binding domain found in conventional CAR molecules. The CAR construct targets the pMHC complex corresponding to the original TCR molecule and activates T cells expressing the CAR via the intracellular signaling domain of the CAR molecule, thereby achieving the desired immunological effect (e.g., target cell killing).

[0067] In some embodiments, the TCR molecule (or the variable regions of the α and β chains or binding domains comprising these) may be linked to a binder to form an immune complex, and the binder may be, for example, a detectable tag, a therapeutic agent, a tracer, etc.

[0068] In some embodiments, the TCR molecule (or the variable regions of the α and β chains or binding domains thereof) can be linked to a protein tag to form a fusion protein. The protein tag may include a purification tag and a detectable tag. Purification tags include, but are not limited to, His6 tag, Flag tag, MBP tag, GST tag, SUMO tag, etc. The detectable tag can be used to indicate the presence or content of the TCR molecule (or the variable regions of the α and β chains or binding domains thereof) in a sample, or to track the location of the TCR molecule (or the variable regions of the α and β chains or binding domains thereof) within a subject's body or cells. Examples of detectable tags include various enzymes that can be used in immunodetection, such as horseradish peroxidase (HRP) and alkaline phosphatase (ALP), and fluorescent proteins, such as GFP. Due to the specific binding ability of the TCR molecule (or the variable regions of the α and β chains or the binding domains composed thereof) to the corresponding pMHC complex, the amount of the TCR molecule (or the variable regions of the α and β chains or the binding domains composed thereof) can be determined from the amount of detectable tag linked to the TCR molecule (or the variable regions of the α and β chains or the binding domains composed thereof), and the content of the corresponding pMHC complex in a sample can be determined. When the corresponding pMHC complex is useful as a tumor marker, the fusion protein can be used for the detection and diagnosis of tumors.

[0069] In some other embodiments, the TCR molecules provided herein (or the variable regions of the α and β chains or binding domains thereof) can be linked to a cytokine or therapeutic protein to form a fusion protein. In such cases, the ability of the TCR molecules (or the variable regions of the α and β chains or binding domains thereof) to specifically bind to the corresponding pMHC complex allows the cytokine or therapeutic protein to be delivered to a specific tissue or cell for a specific purpose, thereby achieving the therapeutic effect of the cytokine or therapeutic protein.

[0070] In some embodiments, the TCR molecule (or the variable regions of the α and β chains or binding domains composed thereof) can be linked to a chemotherapeutic agent, such as asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, gemcitabine, hydroxyurea, methotrexate, taxol, rituximab, vinblastine, or vincristine, thereby enabling the chemotherapeutic agent to be delivered to specific cells or tissues via the antigen specificity of the mTCR molecule.

[0071] In some embodiments, the TCR molecule (or the variable regions of the α and β chains or binding domains thereof) is irradiated with a radioisotope (e.g., 3 H, 14 C. 35 S) can be used for tracking or treatment purposes.

[0072] Host cells expressing TCR molecules Furthermore, the present specification provides host cells that express the TCR molecules provided herein (or the variable regions of the α chain and β chain, or binding domains comprising these).

[0073] The host cell used refers to any type of cell capable of expressing the TCR molecule (or the α and β chain variable regions or binding domains consisting of these) provided herein. Preferably, the host cell expresses the complete TCR molecule, i.e., including the α and β chain variable regions, the α and β chain constant regions, the transmembrane region, and the intracellular region. The host cell may be a eukaryotic or prokaryotic cell. Preferably, the host cell is a mammalian cell. Most preferably, the host cell is a human cell. The host cell may be of any cell type, derived from any type of tissue, and at any stage of development, but is preferably a peripheral blood lymphocyte (PBL) or peripheral blood mononuclear cell (PBMC). More preferably, the host cell is a T cell. For purposes herein, a T cell can be any T cell, e.g., a cultured T cell, e.g., a primary T cell or a T cell from a cultured T cell line, e.g., Jurkat, SupTl, etc., or a T cell obtained from a mammal. If obtained from a mammal, the T cell can be obtained from a number of sources, including, but not limited to, blood, bone marrow, lymph nodes, thymus, or other tissues or fluids. The T cell can be enriched or purified. The T cell can be any type of T cell and at any stage of development, including, but not limited to, CD4+ / CD8+ double-positive T cells, CD4+ helper T cells, e.g., Th1 and Th2 cells, CD4+ T cells, CD8+ T cells (e.g., cytotoxic T cells), tumor-infiltrating lymphocytes (TILs), memory T cells (e.g., central memory T cells and effector memory T cells), naive T cells, etc.

[0074] Additionally, the present specification provides a host cell that expresses a CAR provided herein. Preferably, the host cell is a human cell, particularly a human T cell or NK cell.

[0075] The effector function of host cells (e.g., T cells) expressing a TCR or CAR can be detected by many means. In some embodiments, host cells (e.g., T cells) expressing a TCR or CAR are incubated with target cells, and the number or activity of the target cells is detected (e.g., by detecting LDH release), which reflects the ability of the host cells to kill the target cells. In some embodiments, host cells (e.g., T cells) expressing a TCR or CAR are incubated with target cells, and the amount of IFN-γ secreted by the host cells or the expression of CD69 is detected, which evaluates the immune effect of the host cells.

[0076] Drug Compositions and Methods of Treatment The TCR molecules (or the variable regions of the α chain and β chain or binding domains composed thereof) provided herein and fusion proteins or immunoconjugates comprising the TCR molecules (or the variable regions of the α chain and β chain or binding domains composed thereof) can be formulated into pharmaceutical compositions together with a pharmaceutically acceptable carrier and administered to a subject to prevent or treat tumors.

[0077] In some embodiments, the TCR molecules (or the variable regions of the α and β chains or binding domains comprising these) and fusion proteins or immunoconjugates comprising the TCR molecules (or the variable regions of the α and β chains or binding domains comprising these) provided herein can be administered in combination with one or more other drugs (e.g., anti-tumor agents).

[0078] In some embodiments, the disease or condition to be treated is a tumor, particularly a tumor expressing the antigenic peptide VVGAVGVGK (SEQ ID NO: 91) or VVVGAVGVGK (SEQ ID NO: 92). These tumors include acute lymphocytic cancer, acute myeloid leukemia, alveolar rhabdomyosarcoma, bone cancer, brain cancer, breast cancer, anal cancer, anal canal cancer or anorectal cancer, eye cancer, intrahepatic bile duct cancer, joint cancer, cervical cancer, gallbladder cancer or pleural cancer, nasal cancer, nasal cavity cancer or middle ear cancer, oral cancer, vaginal cancer, vulvar cancer, chronic lymphocytic leukemia, chronic myeloid carcinoma, colon cancer, colorectal cancer, endometrial cancer, esophageal cancer, cervical cancer, gastrointestinal cancer / tumor, glioma, hominis cancer, and the like. Cancers include, but are not limited to, Hodgkin's lymphoma, hypopharyngeal cancer, renal cancer, laryngeal cancer, liver cancer, lung cancer, malignant mesothelioma, melanoma, multiple myeloma, nasopharyngeal cancer, non-Hodgkin's lymphoma, oropharyngeal cancer, ovarian cancer, penile cancer, pancreatic cancer, peritoneal cancer, peritoneal mesothelial and mesenteric cancer, pharyngeal cancer, prostate cancer, rectal cancer, kidney cancer, skin cancer, small intestine cancer, soft tissue cancer, stomach cancer, testicular cancer, thyroid cancer, uterine cancer, ureteral cancer, and bladder cancer. Suitable cancers include pancreatic cancer, colorectal cancer, lung cancer, endometrial cancer, ovarian cancer, and prostate cancer.

[0079] Nucleic acid molecules encoding the TCR molecules (or the variable regions of the α and β chains or binding domains composed thereof) and fusion proteins or immunoconjugates comprising the TCR molecules (or the variable regions of the α and β chains or binding domains composed thereof) provided herein, expression vectors containing the nucleic acid molecules, and host cells (T cells, CAR cells) transfected with the nucleic acid molecules or expression vectors can also be used for the above-mentioned therapeutic purposes by various means. For example, the expression vector can be introduced into the body of a subject by gene therapy methods known in the art to express the desired protein or polypeptide (e.g., the TCR molecules (or the variable regions of the α and β chains or binding domains composed thereof) and fusion proteins or immunoconjugates comprising the TCR molecules (or the variable regions of the α and β chains or binding domains composed thereof) provided herein, thereby achieving therapeutic purposes.

[0080] The effective dose for such use depends on the severity of the disease and the overall state of the patient's own immune system. The administration regimen will depend on the state of the disease and the condition of the subject, but generally ranges from a single bolus or continuous infusion to multiple doses per day (e.g., every 4-6 hours). A clinician skilled in the art can easily determine whether a subject is a candidate for such treatment, for example, from clinical tests, physical examination, and the subject's family history.

[0081] Detection or Treatment Kits The TCR molecules (or α and β chain variable regions or binding domains comprising these) provided herein and fusion proteins or immune complexes comprising TCR molecules (or α and β chain variable regions or binding domains comprising these) can specifically bind to the corresponding pMHC complexes in a sample. By detecting the amount of ternary complex (i.e., TCR-antigen peptide, MHC molecule) formed, the content (or presence or absence) of the corresponding pMHC complex in a sample can be conveniently determined.

[0082] As described above, for this purpose, the TCR molecules provided herein (or the variable regions of the α and β chains or binding domains comprising them) can be linked to various detection tags so that they can be detected by various means, including, but not limited to, bioluminescence, fluorescence, radioactive labeling, and the amount of product produced by an enzyme-catalyzed reaction. After detecting the content of the corresponding pMHC complex in a sample, the content can be compared with the normal content of the corresponding pMHC complex in a group of healthy individuals and used to determine the status or severity of the disease in the subject from whom the sample was provided. Changes in the content of the corresponding pMHC complex can be repeatedly detected over time during the course of treatment of the subject and used to determine the effectiveness of the treatment, thereby providing a basis for modifying the treatment plan.

[0083] The TCR molecules (or the variable regions of the α and β chains or binding domains comprising these) and fusion proteins or immunoconjugates comprising the TCR molecules (or the variable regions of the α and β chains or binding domains comprising these) provided herein, or host cells, can be contained in a container to form a detection or treatment kit. These containers may be in the form of a box, an ampoule, a vial, a tube, a bag, or any other suitable container known in the art. These containers can be made of plastic, glass, laminated paper, metal foil, or other materials suitable for storing drugs. If necessary, the container provided may further include instructions for use. The instructions typically include information on how to use the TCR molecule (or the α and β variable regions or binding domains comprising these), compositions comprising the TCR molecule (or the α and β variable regions or binding domains comprising these), or host cells to treat or prevent tumors, and may include, for example, a description of the therapeutic agent (e.g., the TCR molecule (or the α and β variable regions or binding domains comprising these), nucleic acid molecule, host cell, etc.), dosage regimens for treating or preventing tumor formation, precautions, warnings, indications, contraindications, adverse reactions, animal pharmacology, clinical studies, and / or reference materials. The instructions can be printed directly on the container (if any), or can be a label affixed to the container, or a separate paper, booklet, card, or foldout provided in or with the container.

[0084] Materials and reagents: 1. PBMCs from healthy individuals 2. Target polypeptide: A*11:01 KRAS-G12V-9 peptide: VVGAVGVGK (SEQ ID NO: 91) A*11:01 KRAS-G12V-10 peptide: VVVGAVGVGK (SEQ ID NO: 92) 3. Brilliant Violet 510® anti-human CD3 antibody: Biolegend 300448 4. APC anti-human CD8a antibody: Biolegend 300912 5. PE-KRAS-G12V-pentamer: PROIMMUNE 6. Chromium Next GEM Single Cell V(D)J Reagent Kit v1.1 7. Lentiviral Vectors 8. Polybrene viral enhancer: Merck Millipore TR-1003-G 9. Anti-CD3 / CD28 magnetic beads: Novoprotein GMP-B038 10. X-VIVO 15:lonza 04418q 11. OKM-100: CansBio sj18 12. RPMI 1640:gibco 11875-093 13. LDH Kit: dojindo 14. Human IFN-γ Kit: Excell Bio EH008-96 15. IFN-γ ELISPOT:Mabtech 3420-4AST-10 16. APC-anti-human CD69 antibody: biolegend 310910

[0085] equipment equipment Flow cytometer: BD LSRFortessa Flow cell sorter: SONY SH800S Microplate Reader: Molecular Devices SpectraMax i3x ELISPOT imaging device: Bio-sys Bioreader BIO-SYS Bioreader 6000-Eβ

[0086] Graphing and Analysis Software FlowJo Graphpad Prism 8.0.1

[0087] Example T cells from three healthy donors were stimulated with dendritic cells carrying the mutant KRAS G12V nonapeptide (VVGAVGVGK) (SEQ ID NO: 91). The proportion of A*11:01-KRAS G12V-specific T cells was detected using pentamer staining and a BD LSRFortessa flow cytometer. Specific T cells were further enriched and selected using a Sony SH800S flow cell sorter and analyzed using FlowJo software. The resulting flow cytometry graph is shown in Figure 1. The enriched cells were subjected to 10x single-cell sequencing to obtain the paired TCR α and TCR β chains.

[0088] Lentiviral vectors containing the TCR genes of the present invention were constructed and used to transduce T cells. T cells transduced with different TCRs were co-incubated with (1) HLA-A*11:01-expressing K562 cells bearing different peptides (G12V-9 peptide: VVGAVGVGK (SEQ ID NO: 91), G12V-10 peptide: VVVGADGVGK (SEQ ID NO: 92), and WT peptide: VVGAGGVGK (SEQ ID NO: 93) + VVVGAGGVGK (SEQ ID NO: 94)) and (2) HLA-A*11:01-expressing BxPC3 cells, HLA-A*11:01 and KRAS-G12V-coexpressing BxPC3 cells, and HLA-A*11:01 and KRAS-G12D-coexpressing BxPC3 cells (effector cells:target cells = 5:1) for 16 h. The killing ability of the different T cells against each target cell was evaluated by detecting LDH release. Data statistics were calculated using Graphpad Prism as shown in Figures 2A and 2B. Using the 8.0.1 software, T cells transduced with different TCRs effectively and specifically killed both A*11:01-expressing KRAS G12V-9 or KRAS G12V-10 peptide-bearing target cells and KRAS G12V-overexpressing target cells.

[0089] As shown in Figure 3, the release of IFN-γ after stimulation of T cells transduced with a given TCR with the KRAS G12V-9 peptide (VVGAVGVGK) (SEQ ID NO: 91) (effector cells:target cells = 1:1) showed that primary T cells expressing the disclosed exemplary TCRs produced large amounts of IFN-γ upon target stimulation alone.

[0090] T cells transduced with a given TCR were co-incubated with target cells bearing different concentrations of the KRAS G12V-9 peptide (VVGAVGVGK) (SEQ ID NO: 91), and the EC50 of different TCRs was evaluated based on the number of spots of IFN-γ release and CD69 expression. As shown in Figures 4A and 4B, the EC50 concentrations of TCRs recognizing the KRAS G12V-9 peptide ranged from 1 nM to 10 nM. As shown in Figure 4C, the EC50 concentrations of TCR1 recognizing the KRAS G12V-9 peptide and the decapeptide differed by approximately 10-fold, with the nonapeptide at 2.24 nM and the decapeptide at 31.3 nM.

[0091] Some amino acid sequences referred to herein are as follows: Amino acid sequence of the α chain of TCR1 MRQVARVIVFLTLSTLSLAKTTQPISMDSYEGQEVNITCSHNNIATNDYITWYQQFPSQGPRFIIQGYKTKVTNEVASLFIPADRKSSTLSLPRVSLSDTAVYYCLVGALSSSWYGQNFVFGPGTRLSVLPYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 1) Amino acid sequence of the α chain V region of TCR1 LAKTTQPISMDSYEGQEVNITCSHNNIATNDYITWYQQFPSQGPRFIIQGYKTKVTNEVASLFIPADRKSSTLSLPRVSLSDTAVYYCLVGALSSSWYGQNFVFGPGTRLSVLP (SEQ ID NO: 2) Amino acid sequence of TCR1 α chain CDR1 NIATNDY (SEQ ID NO: 3) Amino acid sequence of TCR1 α chain CDR2 GYKTK (SEQ ID NO: 4) Amino acid sequence of TCR1 α chain CDR3 LVGALSSSWYGQNFV (SEQ ID NO: 5) Amino acid sequence of the TCR1 β chain MGCRLLCCVVFCLLQAGPLDTAVSQTPKYLVTQMGNDKSIKCEQNLGHDTMYWYKQDSKKFLKIMFSYNNKELIINETVPNRFSPKSPDKAHLNLHINSLELGDSAVYFCASSQSGAQDGYTFGSGTRLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF (SEQ ID NO: 6) Amino acid sequence of the V region of the β chain of TCR1 DTAVSQTPKYLVTQMGNDKSIKCEQNLGHDTMYWYKQDSKKFLKIMFSYNNKELIINETVPNRFSPKSPDKAHLNLHINSLELGDSAVYFCASSQSGAQDGYTFGSGTRLTVV (SEQ ID NO: 7) Amino acid sequence of CDR1 of the β chain of TCR1 LGHDT (SEQ ID NO: 8) Amino acid sequence of CDR2 of the β chain of TCR1 YNNKEL (SEQ ID NO: 9) Amino acid sequence of CDR3 of the β chain of TCR1 ASSQSGAQDGYT (SEQ ID NO: 10) Amino acid sequence of the α chain of TCR2 MAMLLGASVLILWLQPDWVNSQQKNDDQQVKQNSPSLSVQEGRISILNCDYTNSMFDYFLWYKKYPAEGPTFLISISSIKDKNEDGRFTVFLNKSAKHLSLHIVPSQPGDSAVYFCAAKDNAGNMLTFGGGTRLMVKPHIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 11) Amino acid sequence of the V region of the α chain of TCR2 DQQVKQNSPSLSVQEGRISILNCDYTNSMFDYFLWYKKYPAEGPTFLISISSIKDKNEDGRFTVFLNKSAKHLSLHIVPSQPGDSAVYFCAAKDNAGNMLTFGGGTRLMVKP (SEQ ID NO: 12) Amino acid sequence of CDR1 of the α chain of TCR2 NSMFDY (SEQ ID NO: 13) Amino acid sequence of CDR2 of the α chain of TCR2 ISSIKDK (SEQ ID NO: 14) Amino acid sequence of CDR3 of the α chain of TCR2 AAKDNAGNMLT (SEQ ID NO: 15) Amino acid sequence of the β chain of TCR2 MGSRLLCWVLLCLLGAGPVKAGVTQTPRYLIKTRGQQVTLSCSPISGHRSVSWYQQTPGQGLQFLFEYFSETQRNKGNFPGRFSGRQFSNSRSEMNVSTLELGDSALYLCASSLGTAEAFFGQGTRLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF (SEQ ID NO: 16) Amino acid sequence of the V region of the β chain of TCR2 KAGVTQTPRYLIKTRGQQVTLSCSPISGHRSVSWYQQTPGQGLQFLFEYFSETQRNKGNFPGRFSGRQFSNSRSEMNVSTLELGDSALYLCASSLGTAEAFFGQGTRLTVV (SEQ ID NO: 17) Amino acid sequence of CDR1 of the β chain of TCR2 SGHRS (SEQ ID NO: 18) Amino acid sequence of CDR2 of the β chain of TCR2 YFSETQ (SEQ ID NO: 19) Amino acid sequence of CDR3 of the β chain of TCR2 ASSLGTAEAF (SEQ ID NO: 20) Amino acid sequence of the α chain of TCR3 MAMLLGASVLILWLQPDWVNSQQKNDDQQVKQNSPSLSVQEGRISILNCDYTNSMFDYFLWYKKYPAEGPTFLISISSIKDKNEDGRFTVFLNKSAKHLSLHIVPSQPGDSAVYFCAANSGYALNFGKGTSLLVTPHIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 21) Amino acid sequence of the V region of the α chain of TCR3 DQQVKQNSPSLSVQEGRISILNCDYTNSMFDYFLWYKKYPAEGPTFLISISSIKDKNEDGRFTVFLNKSAKHLSLHIVPSQPGDSAVYFCAANSGYALNFGKGTSLLVTP (SEQ ID NO: 22) Amino acid sequence of CDR1 of the α chain of TCR3 NSMFDY (SEQ ID NO: 23) Amino acid sequence of CDR2 of the α chain of TCR3 ISSIKDK (SEQ ID NO: 24) Amino acid sequence of TCR3 α chain CDR3 AANSGYALN (SEQ ID NO: 25) Amino acid sequence of the β chain of TCR3 MGSRLLCWVLLCLLGAGPVKAGVTQTPRYLIKTRGQQVTLSCSPISGHRSVSWYQQTPGQGLQFLFEYFSETQRNKGNFPGRFSGRQFSNSRSEMNVSTLELGDSALYLCASSLGGVGNEQFFGPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG (SEQ ID NO: 26) Amino acid sequence of the V region of the β chain of TCR3 KAGVTQTPRYLIKTRGQQVTLSCSPISGHRSVSWYQQTPGQGLQFLFEYFSETQRNKGNFPGRFSGRQFSNSRSEMNVSTLELGDSALYLCASSLGGVGNEQFFGPGTRLTVL (SEQ ID NO: 27) Amino acid sequence of CDR1 of the β chain of TCR3 SGHRS (SEQ ID NO: 28) Amino acid sequence of CDR2 of the β chain of TCR3 YFSETQ (SEQ ID NO: 29) Amino acid sequence of CDR3 of the β chain of TCR3 ASSLGGVGNEQF (SEQ ID NO: 30) Amino acid sequence of the α chain of TCR4 MMKSLRVLLVILWLQLSWVWSQQKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSGKSPELIMFIYSNGDKEDGRFTAQLNKASQYVSLLIRDSQPSDSATYLCAVERGSTLGRLYFGRGTQLTVWPDIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 31) Amino acid sequence of the V region of the α chain of TCR4 QKEVEQNSGPLSVPEGAIASLNCTYSDRGSQSFFWYRQYSGKSPELIMFIYSNGDKEDGRFTAQLNKASQYVSLLIRDSQPSDSATYLCAVERGSTLGRLYFGRGTQLTVWP (SEQ ID NO: 32) Amino acid sequence of CDR1 of the α chain of TCR4 DRGSQS (SEQ ID NO: 33) Amino acid sequence of CDR2 of the α chain of TCR4 IYSNGD (SEQ ID NO: 34) Amino acid sequence of CDR3 of the α chain of TCR4 AVERGSTLGRLY (SEQ ID NO: 35) Amino acid sequence of the TCR4 β chain MGTSLLCWVVLGFLGTDSVSTDHTGAGVSQSPRYKVTKRGQDVTLRCDPISSHATLYWYQQALGQGPEFLTYFNYEAQPDKSGLPSDRFSAERPEGSISTLTIQRTEQRDSAMYRCASSLSSPTGGPINEQFFGPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG (SEQ ID NO: 36) Amino acid sequence of the V region of the β chain of TCR4 GAGVSQSPRYKVTKRGQDVTLRCDPISSHATLYWYQQALGQGPEFLTYFNYEAQPDKSGLPSDRFSAERPEGSISTLTIQRTEQRDSAMYRCASSLSSPTGGPINEQFFGPGTRLTVL (SEQ ID NO: 37) Amino acid sequence of CDR1 of the β chain of TCR4 SSHAT (SEQ ID NO: 38) Amino acid sequence of CDR2 of the β chain of TCR4 FNYEAQ (SEQ ID NO: 39) Amino acid sequence of CDR3 of the β chain of TCR4 ASSLSSPTGGPINEQF (SEQ ID NO: 40) Amino acid sequence of the α chain of TCR5 MKTFAGFSFLFLWLQLDCMSRGEDVEQSLFLSVREGDSSVINCTYTDSSSTYLYWYKQEPGAGLQLLTYIFSNMDMKQDQRLTVLLNKKDKHLSLRIADTQTGDSAIYFCAESPGGGADGLTFGKGTHLIIQPYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 41) Amino acid sequence of the V region of the α chain of TCR5 GEDVEQSLFLSVREGDSSVINCTYTDSSSTYLYWYKQEPGAGLQLLTYIFSNMDMKQDQRLTVLLNKKDKHLSLRIADTQTGDSAIYFCAESPGGGADGLTFGKGTHLIIQP (SEQ ID NO: 42) Amino acid sequence of CDR1 of the α chain of TCR5 DSSSTY (SEQ ID NO: 43) Amino acid sequence of CDR2 of the α chain of TCR5 IFSNMDM (SEQ ID NO: 44) Amino acid sequence of CDR3 of the α chain of TCR5 AESPGGGADGLT (SEQ ID NO: 45) Amino acid sequence of the TCR5 β chain MLLLLLLLGPGSGLGAVVSQHPSWVICKSGTSVKIECRSLDFQATTMFWYRQFPKQSLMLMATSNEGSKATYEQGVEKDKFLINHASLTLSTLTVTSAHPEDSSFYICSAPRGWAAGVYGYTFGSGTRLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF (SEQ ID NO: 46) Amino acid sequence of the V region of the β chain of TCR5 GAVVSQHPSWVICKSGTSVKIECRSLDFQATTMFWYRQFPKQSLMLMATSNEGSKATYEQGVEKDKFLINHASLTLSTLTVTSAHPEDSSFYICSAPRGWAAGVYGYTFGSGTRLTVV (SEQ ID NO: 47) Amino acid sequence of CDR1 of the β chain of TCR5 DFQATT (SEQ ID NO: 48) Amino acid sequence of CDR2 of the β chain of TCR5 SNEGSKA (SEQ ID NO: 49) Amino acid sequence of CDR3 of the β chain of TCR5 SAPRGWAAGVYGYT (SEQ ID NO: 50) Amino acid sequence of the α chain of TCR6 MLLELIPLLGIHFVLRTARAQSVTQPDIHITVSEGASLELRCNYSYGATPYLFWYVQSPGQGLQLLLKYFSGDTLVQGIKGFEAEFKRSQSSFNLRKPSVHWSDAAEYFCAVGASREYGNKLVFGAGTILRVKSYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 51) Amino acid sequence of the V region of the α chain of TCR6 AQSVTQPDIHITVSEGASLELRCNYSYGATPYLFWYVQSPGQGLQLLLKYFSGDTLVQGIKGFEAEFKRSQSSFNLRKPSVHWSDAAEYFCAVGASREYGNKLVFGAGTILRVKS (SEQ ID NO: 52) Amino acid sequence of CDR1 of the α chain of TCR6 YGATPY (SEQ ID NO: 53) Amino acid sequence of CDR2 of the α chain of TCR6 YFSGDTLV (SEQ ID NO:54) Amino acid sequence of CDR3 of the α chain of TCR6 AVGASREYGNKLV (SEQ ID NO: 55) Amino acid sequence of the TCR6 β chain MDTRVLCCAVICLLGAGLSNAGVMQNPRHLVRRRGQEARLRCSPMKGHSHVYWYRQLPEEGLKFMVYLQKENIIDESGMPKERFSAEFPKEGPSILRIQQVVRGDSAAYFCASSPGGLLHFGNGTRLTVTEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF (SEQ ID NO: 56) Amino acid sequence of the V region of the β chain of TCR6 NAGVMQNPRHLVRRRGQEARLRCSPMKGHSHVYWYRQLPEEGLKFMVYLQKENIIDESGMPKERFSAEFPKEGPSILRIQQVVRGDSAAYFCASSPGGLLHFGNGTRLTVT (SEQ ID NO: 57) Amino acid sequence of CDR1 of the β chain of TCR6 KGHSH (SEQ ID NO: 58) Amino acid sequence of CDR2 of the β chain of TCR6 LQKENI (SEQ ID NO: 59) Amino acid sequence of CDR3 of the β chain of TCR6 ASSPGGLLH (SEQ ID NO: 60) Amino acid sequence of the α chain of TCR7 MAMLLGASVLILWLQPDWVNSQQKNDDQQVKQNSPSLSVQEGRISILNCDYTNSMFDYFLWYKKYPAEGPTFLISISSIKDKNEDGRFTVFLNKSAKHLSLHIVPSQPGDSAVYFCAGKTNTGNQFYFGTGTSLTVIPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 61) Amino acid sequence of the V region of the α chain of TCR7 DQQVKQNSPSLSVQEGRISILNCDYTNSMFDYFLWYKKYPAEGPTFLISISSIKDKNEDGRFTVFLNKSAKHLSLHIVPSQPGDSAVYFCAGKTNTGNQFYFGTGTSLTVIP (SEQ ID NO: 62) Amino acid sequence of CDR1 of the α chain of TCR7 NSMFDY (SEQ ID NO: 63) Amino acid sequence of CDR2 of the α chain of TCR7 ISSIKDK (SEQ ID NO: 64) Amino acid sequence of CDR3 of the α chain of TCR7 AGKTNTGNQFY (SEQ ID NO: 65) Amino acid sequence of the TCR7 β chain MGSRLLCWVLLCLLGAGPVKAGVTQTPRYLIKTRGQQVTLSCSPISGHRSVSWYQQTPGQGLQFLFEYFSETQRNKGNFPGRFSGRQFSNSRSEMNVSTLELGDSALYLCASSSGHPEAFFGQGTRLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF (SEQ ID NO: 66) Amino acid sequence of the V region of the β chain of TCR7 KAGVTQTPRYLIKTRGQQVTLSCSPISGHRSVSWYQQTPGQGLQFLFEYFSETQRNKGNFPGRFSGRQFSNSRSEMNVSTLELGDSALYLCASSSGHPEAFFGQGTRLTVV (SEQ ID NO: 67) Amino acid sequence of CDR1 of the β chain of TCR7 SGHRS (SEQ ID NO: 68) Amino acid sequence of CDR2 of the β chain of TCR7 YFSETQ (SEQ ID NO: 69) Amino acid sequence of CDR3 of the TCR7 β chain ASSSGHPEAF (SEQ ID NO: 70) Amino acid sequence of the α chain of TCR8 METLLGVSLVILWLQLARVNSQQGEEDPQALSIQEGENATMNCSYKTSINNLQWYRQNSGRGLVHLILIRSNEREKHSGRLRVTLDTSKKSSSLLITASRAADTASYFCATDAGGGADGLTFGKGTHLIIQPYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 71) Amino acid sequence of the V region of the α chain of TCR8 SQQGEEDPQALSIQEGENATMNCSYKTSINNLQWYRQNSGRGLVHLILIRSNEREKHSGRLRVTLDTSKKSSSLLITASRAADTASYFCATDAGGGADGLTFGKGTHLIIQP (SEQ ID NO: 72) Amino acid sequence of CDR1 of the α chain of TCR8 TSINN (SEQ ID NO: 73) Amino acid sequence of CDR2 of the α chain of TCR8 IRSNERE (SEQ ID NO: 74) Amino acid sequence of CDR3 of the α chain of TCR8 ATDAGGGADGLT (SEQ ID NO: 75) Amino acid sequence of the β chain of TCR8 MGFRLLCCVAFCLLGAGPVDSGVTQTPKHLITATGQRVTLRCSPRSGDLSVYWYQQSLDQGLQFLIQYYNGEERAKGNILERFSAQQFPDLHSELNLSSLELGDSALYFCASSESRDSGNTIYFGEGSWLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF (SEQ ID NO: 76) Amino acid sequence of the V region of the β chain of TCR8 DSGVTQTPKHLITATGQRVTLRCSPRSGDLSVYWYQQSLDQGLQFLIQYYNGEERAKGNILERFSAQQFPDLHSELNLSSLELGDSALYFCASSESRDSGNTIYFGEGSWLTVV (SEQ ID NO: 77) Amino acid sequence of CDR1 of the β chain of TCR8 SGDLS (SEQ ID NO: 78) Amino acid sequence of CDR2 of the β chain of TCR8 YYNGEE (SEQ ID NO:79) Amino acid sequence of CDR3 of the β chain of TCR8 ASSESRDSGNTIY (SEQ ID NO: 80) Amino acid sequence of the α chain of TCR9 METLLGVSLVILWLQLARVNSQQGEEDPQALSIQEGENATMNCSYKTSINNLQWYRQNSGRGLVHLILIRSNEREKHSGRLRVTLDTSKKSSSLLITASRAADTASYFCATDSGGGADGLTFGKGTHLIIQPYIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 81) Amino acid sequence of the α chain V region of TCR9 SQQGEEDPQALSIQEGENATMNCSYKTSINNLQWYRQNSGRGLVHLILIRSNEREKHSGRLRVTLDTSKKSSSLLITASRAADTASYFCATDSGGGADGLTFGKGTHLIIQP (SEQ ID NO: 82) Amino acid sequence of TCR9 α chain CDR1 TSINN (SEQ ID NO: 83) Amino acid sequence of TCR9 α chain CDR2 IRSNERE (SEQ ID NO: 84) Amino acid sequence of TCR9 α chain CDR3 ATDSGGGADGLT (SEQ ID NO: 85) Amino acid sequence of the β chain of TCR9 MGFRLLCCVAFCLLGAGPVDSGVTQTPKHLITATGQRVTLRCSPRSGDLSVYWYQQSLDQGLQFLIQYYNGEERAKGNILERFSAQQFPDLHSELNLSSLELGDSALYFCASTPGRDSGNTIYFGEGSWLTVVEDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDF (SEQ ID NO: 86) Amino acid sequence of the V region of the β chain of TCR9 DSGVTQTPKHLITATGQRVTLRCSPRSGDLSVYWYQQSLDQGLQFLIQYYNGEERAKGNILERFSAQQFPDLHSELNLSSLELGDSALYFCASTPGRDSGNTIYFGEGSWLTVV (SEQ ID NO: 87) Amino acid sequence of CDR1 of the β chain of TCR9 SGDLS (SEQ ID NO: 88) Amino acid sequence of CDR2 of the β chain of TCR9 YYNGEE (SEQ ID NO: 89) Amino acid sequence of CDR3 of the β chain of TCR9 ASTPGRDSGNTIY (SEQ ID NO: 90)

Claims

1. 1. A binding protein having antigenic peptide / HLA complex antigen specificity, comprising a binding domain containing a T cell receptor (TCR) alpha chain variable region and a TCR beta chain variable region, the antigenic peptide comprises the amino acid sequence VVGAVGVGK (SEQ ID NO: 91); the HLA is HLA-A*11:01, CDR3 of the α chain variable region comprises an amino acid sequence set forth in SEQ ID NO: 5, 15, 25, 35, 45, 55, 65, 75, or 85, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 5, 15, 25, 35, 45, 55, 65, 75, or 85; A binding protein, wherein the CDR3 of the β chain variable region comprises an amino acid sequence set forth in SEQ ID NO: 10, 20, 30, 40, 50, 60, 70, 80, or 90, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 10, 20, 30, 40, 50, 60, 70, 80, or 90.

2. 2. The binding protein of claim 1, wherein the antigenic peptide is VVGAVGVGK (SEQ ID NO: 91) or VVVGAVGVGK (SEQ ID NO: 92).

3. In the α chain variable region, 1) CDR1 comprises the amino acid sequence set forth in SEQ ID NO:3, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:4, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:5, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:5; 2) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 13, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 14, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 15, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 15; 3) CDR1 comprises the amino acid sequence set forth in SEQ ID NO:23, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:24, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:25, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:25; 4) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 33, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 34, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 35, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 35; 5) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 43, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 44, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 45, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 45; 6) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 53, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 54, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 55, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 55; 7) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 63, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 64, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 65, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 65; 8) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 73, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 74, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 75, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 75; or 9) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 83, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 84, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 85 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 85; In the β chain variable region, 1) CDR1 comprises the amino acid sequence set forth in SEQ ID NO:8, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:9, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:10, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:10; 2) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 18, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 19, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 20, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 20; 3) CDR1 comprises the amino acid sequence set forth in SEQ ID NO:28, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:29, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:30, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:30; 4) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 38, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 39, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 40, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 40; 5) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 48, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 49, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 50, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 50; 6) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 58, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 59, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 60, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 60; 7) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 68, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 69, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 70, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 70; 8) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 78, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 79, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 80, or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 80; or 9) CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 88, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 89, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 90 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 90; The binding protein of claim 1.

4. 1) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO:3, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:4, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:5 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:5; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO:8, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:9, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:10 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:10; 2) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 13, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 14, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 15 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 15; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 18, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 19, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 20 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 20; 3) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO:23, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:24, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:25 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:25; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO:28, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:29, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:30 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:30; 4) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 33, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 34, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 35 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 35; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 38, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 39, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 40 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 40; 5) In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 43, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 44, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 45 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 45; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 48, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 49, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 50 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 50; 6) In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 53, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 54, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 55 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 55; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 58, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 59, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 60 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 60; 7) In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 63, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 64, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 65 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 65; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 68, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 69, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 70 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 70; 8) In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 73, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 74, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 75 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 75; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 78, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 79, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 80 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 80; or 9) In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 83, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 84, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 85 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 85; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 88, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 89, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 90 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:

90. The binding protein according to any one of claims 1 to 3.

5. 1. A binding protein comprising a binding domain containing a T cell receptor (TCR) alpha chain variable region and a TCR beta chain variable region, 1) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO:3, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:4, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:5 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:5; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO:8, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:9, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:10 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:10; 2) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 13, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 14, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 15 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 15; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 18, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 19, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 20 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 20; 3) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO:23, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:24, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:25 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:25; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO:28, CDR2 comprises the amino acid sequence set forth in SEQ ID NO:29, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO:30 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:30; 4) in the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 33, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 34, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 35 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 35; in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 38, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 39, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 40 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 40; 5) In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 43, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 44, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 45 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 45; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 48, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 49, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 50 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 50; 6) In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 53, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 54, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 55 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 55; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 58, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 59, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 60 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 60; 7) In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 63, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 64, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 65 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 65; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 68, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 69, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 70 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 70; 8) In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 73, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 74, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 75 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 75; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 78, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 79, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 80 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 80; or 9) In the α chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 83, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 84, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 85 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 85; and in the β chain variable region, CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 88, CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 89, and CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 90 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:

90. Binding proteins.

6. 1) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO:2 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:2, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO:7 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:7; 2) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 12 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 12, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 17 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 17; 3) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO:22 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:22, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO:27 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:27; 4) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 32 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 32, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 37 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 37; 5) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 42 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 42, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 47 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 47; 6) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 52 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 52, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 57 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 57; 7) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 62 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 62, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 67 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, and more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 67; 8) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 72 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 72, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 77 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 77; or 9) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 82 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 82, and the β chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 87 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 87; A binding protein according to any one of claims 1 to 5.

7. 7. The TCR α chain and the TCR β chain are included, 1) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 1 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 1, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 6 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 6; 2) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 11 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 11, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 16 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 16; 3) the α chain comprises the amino acid sequence set forth in SEQ ID NO:21 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:21, and the β chain comprises the amino acid sequence set forth in SEQ ID NO:26 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO:26; 4) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 31 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 31, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 36 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 36; 5) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 41 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 41, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 46 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 46; 6) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 51 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 51, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 56 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 56; 7) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 61 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 61, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 66 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 66; 8) the α chain comprises the amino acid sequence set forth in SEQ ID NO: 71 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 71, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 76 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 76; or 9) the α chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 81 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 81, and the β chain comprises the amino acid sequence set forth in SEQ ID NO: 86 or an amino acid sequence having at least 80% identity, preferably at least 85% identity, more preferably 90% or 95% identity with the amino acid sequence set forth in SEQ ID NO: 86; A binding protein according to any one of claims 1 to 6.

8. The binding protein according to any one of claims 1 to 7, which is a protein having the following form: 1) TCR molecule; 2) an antibody molecule; or 3) CAR molecule.

9. 9. The binding protein of any one of claims 1 to 8, wherein when T cells expressing the binding protein are incubated with HLA-A*11:01 cells in the presence of the antigenic peptide, or when T cells expressing the binding protein are incubated with HLA-A*11:01 cells expressing the antigenic peptide, the T cells specifically kill the HLA-A*11:01 cells.

10. 10. The binding protein of any one of claims 1 to 9, wherein when T cells expressing the binding protein are incubated with HLA-A*11:01 cells in the presence of the antigenic peptide, or when T cells expressing the binding protein are incubated with HLA-A*11:01 cells expressing the antigenic peptide, the T cells produce INF-γ and / or express CD69, or the amount of INF-γ and / or CD69 expression increases.

11. 11. The binding protein of any of claims 1 to 10, further comprising a binding entity covalently or non-covalently linked to said binding domain, preferably said binding entity being a detectable marker, a radioisotope or a therapeutic agent.

12. An isolated nucleic acid molecule encoding the binding protein, α chain variable region, β chain variable region, α chain, or β chain of any one of claims 1 to 11.

13. 13. A vector comprising the nucleic acid molecule of claim 12.

14. A host cell that expresses a binding protein according to any one of claims 1 to 11, or that contains a nucleic acid molecule according to claim 12 or a vector according to claim 13.

15. 15. The host cell according to claim 14, which is a mammalian cell, preferably a human cell.

16. 16. The host cell of claim 14 or 15, which is a T cell or an NK cell.

17. The host cell according to any one of claims 14 to 16, which has killing activity against HLA-A*11:01 cells expressing the antigen peptide VVGAVGVGK (SEQ ID NO: 91) or VVVGAVGVGK (SEQ ID NO: 92).

18. A pharmaceutical composition comprising: 1) a binding protein according to any one of claims 1 to 11, a nucleic acid molecule according to claim 12, a vector according to claim 13 or a host cell according to any one of claims 14 to 17; and 2) A pharmaceutically acceptable carrier.

19. 18. Use of the binding protein of any one of claims 1 to 11, the nucleic acid molecule of claim 12, the vector of claim 13 or the host cell of any one of claims 14 to 17 in the manufacture of a medicament for treating a tumor expressing said antigenic peptide.

20. 19. A method of treating a tumor in a subject, comprising administering to the subject a therapeutically effective amount of the binding protein of any one of claims 1 to 11, the nucleic acid molecule of claim 12, the vector of claim 13, the cell of any one of claims 14 to 17, or the pharmaceutical composition of claim 18.

21. 21. The use according to claim 19 or the method according to claim 20, wherein the tumor is selected from pancreatic adenocarcinoma, colorectal carcinoma, lung carcinoma, bile duct carcinoma, endometrial carcinoma and ovarian carcinoma.

22. A detection kit comprising the binding protein of any one of claims 1 to 11.

Images