ANTIGEN BINDING PROTEINS SPECIFICALLY BINDING CT45

Abstract

The present invention provides an antigen binding protein specifically binding to a CT45 antigenic peptide that is in a complex with a major histocompatibility complex (MHC) protein, wherein the CT45 antigenic peptide comprises or consists of the amino acid sequence of SEQ ID NO: 138 (KIFEMLEGV) and wherein the antigen binding protein comprises a first polypeptide comprising a variable domain V.sub.A comprising complementarity determining regions CDRa1, CDRa2 and CDRa3 and a second polypeptide comprising a variable domain V.sub.B comprising CDRb1, CDRb2 and CDRb3. Also provided are nucleic acids encoding the antigen binding proteins, vectors comprising the nucleic acids, recombinant cells expressing the antigen binding proteins and pharmaceutical compositions comprising the antigen binding proteins. The invention further provides the antigen binding proteins for use in medicine and a method of producing the antigen binding protein.

Claims

1. An antigen binding protein specifically binding to a CT45 antigenic peptide that is in a complex with a major histocompatibility complex (MHC) protein, wherein the CT45 antigenic peptide comprises or consists of the amino acid sequence of SEQ ID NO: 138 (KIFEMLEGV) and wherein the antigen binding protein comprises a first polypeptide comprising a variable domain V.sub.A comprising complementarity determining regions (CDR) CDRa1, CDRa2 and CDRa3 and a second polypeptide comprising a variable domain V.sub.B comprising CDRb1, CDRb2 and CDRb3, wherein 1) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 14, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 16, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 19, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 21, 2) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 24, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 133, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 75, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 136, 3) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 24, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 63, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 66, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 68, 4) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 90, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 92, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 66, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 96, 5) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 2, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 4, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 8, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 10, 6) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 53, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 55, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 58, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 60, 7) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 71, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 72, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 75, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 77, 8) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 99, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 101, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 75, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 104, 9) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 80, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 82, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 85, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 87, 10) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 107, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 109, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 112, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 114, 11) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 125, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 127, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 112, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 130, or 12) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 117, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 119, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 58, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 122, 13) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 24, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 35, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 38, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 40, 14) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 24, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 26, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 29, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 31, or 15) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 43, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 45, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 48, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 50, wherein the antigen binding protein comprises said CDRa1, CDRa3, CDRb1 and CDRb3 sequence(s) with not more than one, two or three amino acid mutations, wherein each of CDRa1, CDRa3, CDRb1 and/or CDRb3 optionally comprise one, two or three amino acid mutations.

2. The antigen binding protein of claim 1, wherein 1) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 15, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 20 2) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 25, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 76, 3) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 25, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 67, 4) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 91, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 95, 5) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 3, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 9, 6) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 54, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 59, 7) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 15, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 76, 8) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 100, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 76, 9) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 81, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 86, 10) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 108, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 113, 11) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 126, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 113, 12) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 118, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 59, 13) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 25, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 39, 14) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 25, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 30, or 15) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 44, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 49, wherein the antigen binding protein comprises said CDRa2 and CDRb2 sequence(s) with not more than one, two, three or four amino acid mutations, wherein each of CDRa2 and/or CDRb2 optionally comprise one, two, three or four amino acid mutations.

3. The antigen binding protein of claim 1, wherein the antigen binding protein is a TCR, optionally wherein the TCR is selected from the group consisting of an α/β TCR, a γ/δ TCR, a single chain TCR, a membrane-bound TCR, a soluble TCR, a monovalent, bivalent or multivalent TCR, a monospecific, bispecific or multispecific TCR, a functional fragment of a TCR, and a fusion protein or chimeric protein comprising a functional fragment of a TCR, optionally wherein the TCR is an α/β TCR or a γ/δ TCR, optionally an α/β TCR.

4. The antigen binding protein of claim 1, wherein V.sub.A comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 13, 132, 62, 89, 1, 52, 70, 98, 79, 106, 124, 116, 34, 23, and 42, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 13, 132, 62, 89, 1, 52, 70, 98, 79, 106, 124, 116, 34, 23, and 42 and comprising the CDRa1, the CDRa2 and the CDRa3; and wherein V.sub.B comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 18, 135, 65, 94, 7, 57, 74, 103, 84, 111, 129, 121, 37, 28 and 47 or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 18, 135, 65, 94, 7, 57, 74, 103, 84, 111, 129, 121, 37, 28 and 47 and comprising the CDRb1, the CDRb2 and the CDRb3, wherein the CDRa1, the CDRa2, the CDRa3, the CDRb1, the CDRb2 and/or the CDRb3 sequences optionally comprise one, two or three amino acid mutations, optionally amino acid substitutions.

5. The antigen binding protein of claim 1, further comprising a constant domain, wherein the constant domain comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 5, 750, 751, 156, 11, 32 and 157, optionally selected from the group consisting of SEQ ID NO: 5, 750, 751, 11 and 32, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 5, 750, 751, 156, 11, 32 or 157.

6. The antigen binding protein of claim 1, wherein the first polypeptide comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 17, 134, 64, 93, 6, 56, 73, 102, 83, 110, 128, 120, 36, 27, 46 and 158-172, optionally selected from the group consisting of SEQ ID NO:17, 134, 64, 93, 6, 56, 73, 102, 83, 110, 128, 120, 36, 27 and 46, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO:17, 134, 64, 93, 6, 56, 73, 102, 83, 110, 128, 120, 36, 27, 46, or 158-172, and the second polypeptide comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 22, 137, 69, 97, 12, 61, 78 105, 88, 115, 131, 123, 41, 33, 51 and 173-187, optionally selected from the group consisting of SEQ ID NO: 22, 137, 69, 97, 12, 61, 78 105, 88, 115, 131, 123, 41, 33 and 51, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 22, 137, 69, 97, 12, 61, 78 105, 88, 115, 131, 123, 41, 33, 51 or 173-187.

7. The antigen binding protein of claim 1, wherein the antigen binding protein does not significantly bind to at least 1, at least 2, at least 3, at least 4, at least 5, or all similar peptides selected from the group consisting of SEQ ID NO: 146 (SP-05-0001), SEQ ID NO: 147 (SP-05-0002), SEQ ID NO: 148 (SP-05-0003), SEQ ID NO: 149 (SP-05-0004), SEQ ID NO: 150 (SP-05-0005), SEQ ID NO: 151 (SP-05-0006), SEQ ID NO: 152 (SP-05-0007), SEQ ID NO: 153 (SP-05-0008), SEQ ID NO: 154 (SP-05-0009) and SEQ ID NO: 155 (SP-05-0010), optionally from the group consisting of SEQ ID NO: 146 (SP-05-0001), SEQ ID NO: 147 (SP-05-0002), SEQ ID NO: 148 (SP-05-0003), SEQ ID NO: 150 (SP-05-0005), SEQ ID NO: 151 (SP-05-0006), SEQ ID NO: 152 (SP-05-0007), SEQ ID NO: 153 (SP-05-0008), SEQ ID NO: 154 (SP-05-0009) and SEQ ID NO: 155 (SP-05-0010), optionally from the group consisting of SEQ ID NO: 147 (SP-05-0002), SEQ ID NO: 151 (SP-05-0006), SEQ ID NO: 152 (SP-05-0007) and SEQ ID NO: 155 (SP-05-0010).

8. The antigen binding protein of claim 1, wherein the antigen binding protein is capable of activating a CD4+ T cell, in particular a CD4+CD8− T cell, and/or a CD8+ T cell, in particular a CD8+CD4-T cell, and wherein the antigen binding protein is optionally a TCR, optionally an α/β TCR or γ/δ TCR.

9. A nucleic acid comprising a sequence encoding the antigen binding protein of claim 1.

10. A vector comprising the nucleic acid of claim 9.

11. A host cell comprising the antigen binding protein of claim 1.

12. A pharmaceutical composition comprising the antigen binding protein of claim 1.

13. The antigen binding protein of claim 1 for use in medicine, optionally for use in a method of treatment and/or diagnosis of a proliferative disease, in particular cancer.

14. An in-vitro method of detecting cancer, in particular cancer expressing CT45, in a biological sample comprising: a) contacting the biological sample with the antigen binding protein of claim 1, and b) detecting binding of the antigen binding protein to the biological sample.

15. A method of producing the antigen binding protein, comprising a) providing a host cell, b) providing a genetic construct comprising a nucleic acid encoding the antigen binding protein of claim 1, c) introducing the genetic construct into the host cell, and d) expressing the genetic construct by the host cell.

16. The antigen binding protein of claim 2, wherein the antigen binding protein is a TCR, optionally wherein the TCR is selected from the group consisting of an α/β TCR, a γ/δ TCR, a single chain TCR, a membrane-bound TCR, a soluble TCR, a monovalent, bivalent or multivalent TCR, a monospecific, bispecific or multispecific TCR, a functional fragment of a TCR, and a fusion protein or chimeric protein comprising a functional fragment of a TCR, optionally wherein the TCR is an α/β TCR or a γ/δ TCR, optionally an α/β TCR.

17. The antigen binding protein of claim 2, wherein V.sub.A comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 13, 132, 62, 89, 1, 52, 70, 98, 79, 106, 124, 116, 34, 23, and 42, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 13, 132, 62, 89, 1, 52, 70, 98, 79, 106, 124, 116, 34, 23, and 42 and comprising the CDRa1, the CDRa2 and the CDRa3; and wherein V.sub.B comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 18, 135, 65, 94, 7, 57, 74, 103, 84, 111, 129, 121, 37, 28 and 47 or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 18, 135, 65, 94, 7, 57, 74, 103, 84, 111, 129, 121, 37, 28 and 47 and comprising the CDRb1, the CDRb2 and the CDRb3, wherein the CDRa1, the CDRa2, the CDRa3, the CDRb1, the CDRb2 and/or the CDRb3 sequences optionally comprise one, two or three amino acid mutations, optionally amino acid substitutions.

18. The antigen binding protein of claim 3, wherein V.sub.A comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 13, 132, 62, 89, 1, 52, 70, 98, 79, 106, 124, 116, 34, 23, and 42, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 13, 132, 62, 89, 1, 52, 70, 98, 79, 106, 124, 116, 34, 23, and 42 and comprising the CDRa1, the CDRa2 and the CDRa3; and wherein V.sub.B comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 18, 135, 65, 94, 7, 57, 74, 103, 84, 111, 129, 121, 37, 28 and 47 or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 18, 135, 65, 94, 7, 57, 74, 103, 84, 111, 129, 121, 37, 28 and 47 and comprising the CDRb1, the CDRb2 and the CDRb3, wherein the CDRa1, the CDRa2, the CDRa3, the CDRb1, the CDRb2 and/or the CDRb3 sequences optionally comprise one, two or three amino acid mutations, optionally amino acid substitutions.

19. The antigen binding protein of claim 2, further comprising a constant domain, wherein the constant domain comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 5, 750, 751, 156, 11, 32 and 157, optionally selected from the group consisting of SEQ ID NO: 5, 750, 751, 11 and 32, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 5, 750, 751, 156, 11, 32 or 157.

20. The antigen binding protein of claim 3, further comprising a constant domain, wherein the constant domain comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 5, 750, 751, 156, 11, 32 and 157, optionally selected from the group consisting of SEQ ID NO: 5, 750, 751, 11 and 32, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 5, 750, 751, 156, 11, 32 or 157.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0760] FIG. 1: Functional avidity (EC.sub.50) measured by killing efficiency of CT45-IP peptide-loaded T2 cells by TCR-transfected T cells. Constitutively luciferase-expressing T2-cells were loaded with titrated amounts of CT45-IP peptide and then co-cultivated with CD8+ T cells transfected with specific TCRs. Killing was analyzed by measuring luciferase activity in the supernatant which is released by dying T2 cells. The assay was repeated twice with cells of two different donors (indicated by filled circles and diamond-shaped symbols). The functional avidity was assessed by calculating the half maximal killing efficiency of the tested TCRs.

[0761] FIG. 2: Cross-reactivity check for sequence similar-peptides. Constitutively luciferase-expressing T2-cells were loaded with CT45-IP peptide, 10 different sequence-similar peptides, the irrelevant peptide control NYESO1-001 at a concentration of 10 μM per peptide or not loaded, respectively. Those T2 cell were then co-cultivated with CD8+ T cells transfected with specific TCRs. Killing was analyzed by measuring luciferase activity in the supernatant which is released by dying T2 cells. The assay was repeated twice with cells of two different donors (black bars=assay 1, donor 1; red bars=assay 2, donor 2).

[0762] FIG. 3: TCR surface-staining. Flow-cytometric assessment of TCR expression measured by pHLA-Dextramer-binding. Histograms show TCR-mRNA electroporated T cells (black line) and the Mock-TCR control (light grey dotted line) after staining with CT45-IP-HLA-A2*02 dextramers. Percent positive events are indicated in the plots. The Mock-TCR control is used as reference for the dextramer-negative area.

[0763] FIG. 4: Tumor cell line efficacy. Live cell monitoring of tumor cell lines expressing RFP which are co-cultivated with or without T cells expressing our TCRs of interest. Red counts representing the tumor cells were quantified over a period of 48 h and normalized to time point 0 h. Plots on the left side show tumor cell line NCIH1703 and plots on the right side show proliferation of cell line A375. CD8+ T cell electroporated with Mock-TCR (negative control, top), TCR-9 (middle) and TCR-7 (below) are shown. Target cells additionally loaded with 10 μM CT45-IP peptide as positive control were used (circle with spot in the middle) as well as target cells without effector cells (asterisk) and different E:T ratios of effector cells expressing the TCR of interest (circles in different shades of grey).

EXAMPLES

Material and Methods

TCR-Identification

[0764] The alpha and beta TCR chain sequences were isolated from T cells of healthy donors. To ensure enrichment of peptide-specific T cells, the cells were either repeatedly stimulated with artificial antigen-presenting cells coated with CT45-IP-MHC and CD28 (Priming) as described in Walter et al., 2003 J Immunol., November 15; 171(10):4974-8 and subsequently, single-cell sorted using CT45-IP-HLA-A*02 tetramers or alternatively stimulated with CT45-IP-loaded T2 cells. After sufficient expansion, the cells were sorted using CT45-IP-HLA-A*02 tetramers.

[0765] The TCR nucleotide sequences were obtained via standard methods such as 5′ RACE and Sanger sequencing as described e.g. in Molecular Cloning, Laboratory Manual, Fourth Edition by Green and Sambrook. The genes encoding the V regions and J regions of the TCRs are listed in Table 2. The annotation was performed by GeneData 11.0.1 using the IMGT/GENE-DB (Version: 28 Nov. 2019) as reference database. The TCR amino acid sequences are listed in Table 3.

TABLE-US-00002 TABLE 2 Identified TCRs ID Valpha Jalpha Vbeta Jbeta TCR-1 TRAV38-1 TRAJ22 TRBV7-9 TRBJ2-7 TCR-2 TRAV14/DV4 TRAJ52 TRBV13 TRBJ1-1 TCR-3 TRAV14/DV4 TRAJ33 TRBV27 TRBJ1-5 TCR-4 TRAV3 TRAJ30 TRBV6-2 TRBJ2-1 TCR-5 TRAV35 TRAJ26 TRBV9 TRBJ2-7 TCR-6 TRAV12-3 TRAJ7 TRBV4-1 TRBJ2-1 TCR-7 TRAV38-2/DV8 TRAJ32 TRBV13 TRBJ2-1 TCR-8 TRAV19 TRAJ40 TRBV13 TRBJ2-1 TCR-9 TRAV5 TRAJ17 TRBV2 TRBJ2-1 TCR-10 TRAV1-2 TRAJ44 TRBV6-1 TRBJ2-1 TCR-11 TRAV22 TRAJ44 TRBV6-1 TRBJ2-7 TCR-12 TRAV27 TRAJ50 TRBV4-1 TRBJ1-2 TCR-13 TRAV14/DV4 TRAJ5 TRBV19 TRBJ2-1 TCR-14 TRAV14/DV4 TRAJ22 TRBV11-2 TRBJ1-6 TCR-15 TRAV21 TRAJ37 TRBV5-1 TRBJ2-7

TABLE-US-00003 TABLE 3 TCR amino acid sequences SEQ ID Region/ NO: TCR Chain Domian Sequence 1 TCR-5 alpha variable QQLNQSPQSMFIQEGEDVSMNCTSSSIFNTWLWYKQDPGEGP domain VLLIALYKAGELTSNGRLTAQFGITRKDSFLNISASIPSDVGIYFCAG GYNYGQNFVFGPGTRLSVLP 2 TCR-5 alpha CDR1 SIFNT 3 TCR-5 alpha CDR2 LYKAGEL 4 TCR-5 alpha CDR3 AGGYNYGQNFV 5 TCR-5 alpha constant YIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 6 TCR-5 alpha full- QQLNQSPQSMFIQEGEDVSMNCTSSSIFNTWLWYKQDPGEGP length VLLIALYKAGELTSNGRLTAQFGITRKDSFLNISASIPSDVGIYFCAG GYNYGQNFVFGPGTRLSVLPYIQNPDPAVYQLRDSKSSDKSVCLF TDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNK SDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQN LSVIGFRILLLKVAGFNLLMTLRLWSS 7 TCR-5 beta variable GVTQTPKHLITATGQRVTLRCSPRSGDLSVYWYQQSLDQGLQFLI domain QYYNGEERAKGNILERFSAQQFPDLHSELNLSSLELGDSALYFCAS SAGLAGGYEQYFGPGTRLTVT 8 TCR-5 beta CDR1 SGDLS 9 TCR-5 beta CDR2 YYNGEE 10 TCR-5 beta CDR3 ASSAGLAGGY EQY 11 TCR-5 beta constant EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSW domain WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD SRG 12 TCR-5 beta full- GVTQTPKHLITATGQRVTLRCSPRSGDLSVYWYQQSLDQGLQFLI length QYYNGEERAKGNILERFSAQQFPDLHSELNLSSLELGDSALYFCAS SAGLAGGYEQYFGPGTRLTVTEDLKNVFPPEVAVFEPSEAEISHT QKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQP ALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWT QDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGK ATLYAVLVSALVLMAMVKRKDSRG 13 TCR-1 alpha variable QTVTQSQPEM SVQEAETVTL SCTYDTSENN YYLFWYKQPP domain SRQMILVIRQ EAYKQQNATE NRFSVNFQKA AKSFSLKISD SQLGDTAMYF CAPLGLVAGS ARQLTFGSGT QLTVLP 14 TCR-1 alpha CDR1 TSENNYY 15 TCR-1 alpha CDR2 QEAYKQQN 16 TCR-1 alpha CDR3 APLGLVAGSA RQLT 750 TCR-1 alpha constant DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 17 TCR-1 alpha full- QTVTQSQPEM SVQEAETVTL SCTYDTSENN YYLFWYKQPP length SRQMILVIRQ EAYKQQNATE NRFSVNFQKA AKSFSLKISD SQLGDTAMYF CAPLGLVAGS ARQLTFGSGT QLTVLP DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 18 TCR-1 beta variable GVSQDPRHKI TKRGQNVTFR CDPISEHNRL YWYRQTLGQG domain PEFLTYFQNE AQLEKSRLLS DRFSAERPKG SFSTLEIQRT EQGDSAMYLC ASSTDITSYE QYFGPGTRLT VT 19 TCR-1 beta CDR1 SEHNR 20 TCR-1 beta CDR2 FQNEAQ 21 TCR-1 beta CDR3 ASSTDITSYE QY 11 TCR-1 beta constant EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSW domain WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD SRG 22 TCR-1 beta full- GVSQDPRHKI TKRGQNVTFR CDPISEHNRL YWYRQTLGQG length PEFLTYFQNE AQLEKSRLLS DRFSAERPKG SFSTLEIQRT EQGDSAMYLC ASSTDITSYE QYFGPGTRLT VT EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSW WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD SRG 23 TCR-14 alpha variable QKITQTQPGM FVQEKEAVTL DCTYDTSDQS YGLFWYKQPS domain SGEMIFLIYQ GSYDEQNATE GRYSLNFQKA RKSANLVISA SQLGDSAMYF CAINGLPGSA RQLTFGSGTQ LTVLP 24 TCR-14 alpha CDR1 TSDQSYG 25 TCR-14 alpha CDR2 QGSYDEQN 26 TCR-14 alpha CDR3 AINGLPGSAR QLT 750 TCR-14 alpha constant DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 27 TCR-14 alpha full- QKITQTQPGM FVQEKEAVTL DCTYDTSDQS YGLFWYKQPS length SGEMIFLIYQ GSYDEQNATE GRYSLNFQKA RKSANLVISA SQLGDSAMYF CAINGLPGSA RQLTFGSGTQ LTVLP DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 28 TCR-14 beta variable GVAQSPRYKI IEKRQSVAFW CNPISGHATL YWYQQILGQG domain PKLLIQFQNN GVVDDSQLPK DRFSAERLKG VDSTLKIQPA KLEDSAVYLC ASSRFVATGT SPLHFGNGTR LTVT 29 TCR-14 beta CDR1 SGHAT 30 TCR-14 beta CDR2 FQNNGV 31 TCR-14 beta CDR3 ASSRFVATGT SPLH 32 TCR-14 beta constant EDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSW domain WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD F 33 TCR-14 beta full- GVAQSPRYKI IEKRQSVAFW CNPISGHATL YWYQQILGQG length PKLLIQFQNN GVVDDSQLPK DRFSAERLKG VDSTLKIQPA KLEDSAVYLC ASSRFVATGT SPLHFGNGTR LTVT EDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSW WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD F 34 TCR-13 alpha variable QKITQTQPGM FVQEKEAVTL DCTYDTSDQS YGLFWYKQPS domain SGEMIFLIYQ GSYDEQNATE GRYSLNFQKA RKSANLVISA SQLGDSAMYF CAMSDPIMDT GRRALTFGSG TRLQVQP 24 TCR-13 alpha CDR1 TSDQSYG 25 TCR-13 alpha CDR2 QGSYDEQN 35 TCR-13 alpha CDR3 AMSDPIMDTG RRALT 751 TCR-13 alpha constant NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 36 TCR-13 alpha full- QKITQTQPGM FVQEKEAVTL DCTYDTSDQS YGLFWYKQPS length SGEMIFLIYQ GSYDEQNATE GRYSLNFQKA RKSANLVISA SQLGDSAMYF CAMSDPIM DT GRRALTFGSG TRLQVQP NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 37 TCR-13 beta variable GITQSPKYLF RKEGQNVTLS CEQNLNHDAM YWYRQDPGQG domain LRLIYYSQIV NDFQKGDIAE GYSVSREKKE SFPLTVTSAQ KNPTAFYLCA SKSRGPNLAD TQYFGPGTRL TVL 38 TCR-13 beta CDR1 LNHDA 39 TCR-13 beta CDR2 SQIVND 40 TCR-13 beta CDR3 ASKSRGPNLA DTQY 11 TCR-13 beta constant EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSW domain WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD SRG 41 TCR-13 beta full- GITQSPKYLF RKEGQNVTLS CEQNLNHDAM YWYRQDPGQG length LRLIYYSQIV NDFQKGDIAE GYSVSREKKE SFPLTVTSAQ KNPTAFYLCA SKSRGPNLAD TQYFGPGTRL TVL EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSW WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD SRG 42 TCR-15 alpha variable KQEVTQIPAA LSVPEGENLV LNCSFTDSAI YNLQWFRQDP domain GKGLTSLLLI QSSQREQTSG RLNASLDKSS GRSTLYIAAS QPGDSATYLC AVLLTGKLIF GQGTTLQVKP 43 TCR-15 alpha CDR1 DSAIYN 44 TCR-15 alpha CDR2 IQSSQRE 45 TCR-15 alpha CDR3 AVLLTGKLI 750 TCR-15 alpha constant DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 46 TCR-15 alpha full- KQEVTQIPAA LSVPEGENLV LNCSFTDSAI YNLQWFRQDP length GKGLTSLLLI QSSQREQTSG RLNASLDKSS GRSTLYIAAS QPGDSATYLC AVLLTGKLIF GQGTTLQVKP DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 47 TCR-15 beta variable GVTQTPRYLI KTRGQQVTLS CSPISGHRSV SWYQQTPGQG domain LQFLFEYFSE TQRNKGNFPG RFSGRQFSNS RSEMNVSTLE LGDSALYLCA SSGGPGPSGE QYFGPGTRLT VT 48 TCR-15 beta CDR1 SGHRS 49 TCR-15 beta CDR2 YFSETQ 50 TCR-15 beta CDR3 ASSGGPGPSG EQY 11 TCR-15 beta constant EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSW domain WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD SRG 51 TCR-15 beta full- GVTQTPRYLI KTRGQQVTLS CSPISGHRSV SWYQQTPGQG length LQFLFEYFSE TQRNKGNFPG RFSGRQFSNS RSEMNVSTLE LGDSALYLCA SSGGPGPSGE QYFGPGTRLT VT EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSW WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD SRG 52 TCR-6 alpha variable QQKEVEQDPG PLSVPEGAIV SLNCTYSNSA FQYFMWYRQY domain SRKGPELLMY TYSSGNKEDG RFTAQVDKSS KYISLFIRDS QPSDSATYLC AMSGNSGARD YGNNRLAFGK GNQVVVIP 53 TCR-6 alpha CDR1 NSAFQY 54 TCR-6 alpha CDR2 TYSSGN 55 TCR-6 alpha CDR3 AMSGNSGARD YGNNRLA 751 TCR-6 alpha constant NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 56 TCR-6 alpha full- QQKEVEQDPG PLSVPEGAIV SLNCTYSNSA FQYFMWYRQY length SRKGPELLMY TYSSGNKEDG RFTAQVDKSS KYISLFIRDS QPSDSATYLC AMSGNSGARD YGNNRLAFGK GNQVVVIP NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 57 TCR-6 beta variable EVTQTPKHLV MGMTNKKSLK CEQHMGHRAM YWYKQKAKKP domain PELMFVYSYE KLSINESVPS RFSPECPNSS LLNLHLHALQ PEDSALYLCA SATWEEAGPY NEQFFGPGTR LTVL 58 TCR-6 beta CDR1 MGHRA 59 TCR-6 beta CDR2 YSYEKL 60 TCR-6 beta CDR3 ASATWEEAGP YNEQF 11 TCR-6 beta constant EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSW domain WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD SRG 61 TCR-6 beta full- EVTQTPKHLV MGMTNKKSLK CEQHMGHRAM YWYKQKAKKP length PELMFVYSYE KLSINESVPS RFSPECPNSS LLNLHLHALQ PEDSALYLCA SATWEEAGPY NEQFFGPGTR LTVL EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSW WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD SRG 62 TCR-3 alpha variable QKITQTQPGM FVQEKEAVTL DCTYDTSDQS YGLFWYKQPS domain SGEMIFLIYQ GSYDEQNATE GRYSLNFQKA RKSANLVISA SQLGDSAMYF CGYSNYQLIW GAGTKLIIKP 24 TCR-3 alpha CDR1 TSDQSYG 25 TCR-3 alpha CDR2 QGSYDEQN 63 TCR-3 alpha CDR3 GYSNYQLI 750 TCR-3 alpha constant DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 64 TCR-3 alpha full- QKITQTQPGM FVQEKEAVTL DCTYDTSDQS YGLFWYKQPS length SGEMIFLIYQ GSYDEQNATE GRYSLNFQKA RKSANLVISA SQLGDSAMYF CGYSNYQLIW GAGTKLIIKP DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKWLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 65 TCR-3 beta variable QVTQNPRYLI TVTGKKLTVT CSQNMNHEYM SWYRQDPGLG domain LRQIYYSMNV EVTDKGDVPE GYKVSRKEKR NFPLILESPS PNQTSLYFCA SREGTGGYQP QHFGDGTRLS IL 66 TCR-3 beta CDR1 MNHEY 67 TCR-3 beta CDR2 SMNVEV 68 TCR-3 beta CDR3 ASREGTGGYQ PQH 32 TCR-3 beta constant EDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSW domain WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD F 69 TCR-3 beta full- QVTQNPRYLI TVTGKKLTVT CSQNMNHEYM SWYRQDPGLG length LRQIYYSMNV EVTDKGDVPE GYKVSRKEKR NFPLILESPS PNQTSLYFCA SREGTGGYQP QHFGDGTRLS IL EDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSW WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD F 70 TCR-7 alpha variable QTVTQSQPEM SVQEAETVTL SCTYDTSESD YYLFWYKQPP domain SRQMILVIRQ EAYKQQNATE NRFSVNFQKA AKSFSLKISD SQLGDAAMYF CTFRYGGATN KLIFGTGTLL AVQP 71 TCR-7 alpha CDR1 TSESDYY 15 TCR-7 alpha CDR2 QEAYKQQN 72 TCR-7 alpha CDR3 TFRYGGATNK LI 751 TCR-7 alpha constant NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 73 TCR-7 alpha full- QTVTQSQPEM SVQEAETVTL SCTYDTSESD YYLFWYKQPP length SRQMILVIRQ EAYKQQNATE NRFSVNFQKA AKSFSLKISD SQLGDAAMYF CTFRYGGATN KLIFGTGTLL AVQP NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 74 TCR-7 beta variable GVIQSPRHLI KEKRETATLK CYPIPRHDTV YWYQQGPGQD domain PQFLISFYEK MQSDKGSIPD RFSAQQFSDY HSELNMSSLE LGDSALYFCA SRGGWAETPD TQYFGPGTRL TVL 75 TCR-7 beta CDR1 PRHDT 76 TCR-7 beta CDR2 FYEKMQ 77 TCR-7 beta CDR3 ASRGGWAETP DTQY 11 TCR-7 beta constant EDLKNVFPPE VAVFEPSEAE ISHTQKATLV CLATGFYPDH domain VELSWWVNGK EVHSGVSTDP QPLKEQPALN DSRYCLSSRL RVSATFWQNP RNHFRCQVQF YGLSENDEWT QDRAKPVTQI VSAEAWGRAD CGFTSESYQQ GVLSATILYE ILLGKATLYA VLVSALVLMA MVKRKDSRG 78 TCR-7 beta full- GVIQSPRHLI KEKRETATLK CYPIPRHDTV YWYQQGPGQD length PQFLISFYEK MQSDKGSIPD RFSAQQFSDY HSELNMSSLE LGDSALYFCA SRGGWAETPD TQYFGPGTRL TVL EDLKNVFPPE VAVFEPSEAE ISHTQKATLV CLATGFYPDH VELSWWVNGK EVHSGVSTDP QPLKEQPALN DSRYCLSSRL RVSATFWQNP RNHFRCQVQF YGLSENDEWT QDRAKPVTQI VSAEAWGRAD CGFTSESYQQ GVLSATILYE ILLGKATLYA VLVSALVLMA MVKRKDSRG 79 TCR-9 alpha variable EDVEQSLFLS VREGDSSVIN CTYTDSSSTY LYWYKQEPGA domain GLQLLTYIFS NMDMKQDQRL TVLLNKKDKH LSLRIADTQT GDSAIYFCAE KETAGNKLTF GGGTRVLVKP 80 TCR-9 alpha CDR1 DSSSTY 81 TCR-9 alpha CDR2 IFSNMDM 82 TCR-9 alpha CDR3 AEKETAGNKL T 751 TCR-9 alpha constant NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 83 TCR-9 alpha full- EDVEQSLFLS VREGDSSVIN CTYTDSSSTY LYWYKQEPGA length GLQLLTYIFS NMDMKQDQRL TVLLNKKDKH LSLRIADTQT GDSAIYFCAE KETAGNKLTF GGGTRVLVKP NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 84 TCR-9 beta variable EPEVTQTPSH QVTQMGQEVI LRCVPISNHL YFYWYRQILG domain QKVEFLVSFY NNEISEKSEI FDDQFSVERP DGSNFTLKIR STKLEDSAMY FCASTVQSPR TNEQFFGPGT RLTVL 85 TCR-9 beta CDR1 SNHLY 86 TCR-9 beta CDR2 FYNNEI 87 TCR-9 beta CDR3 ASTVQSPRTN EQF 11 TCR-9 beta constant EDLKNVFPPE VAVFEPSEAE ISHTQKATLV CLATGFYPDH domain VELSWWVNGK EVHSGVSTDP QPLKEQPALN DSRYCLSSRL RVSATFWQNP RNHFRCQVQF YGLSENDEWT QDRAKPVTQI VSAEAWGRAD CGFTSESYQQ GVLSATILYE ILLGKATLYA VLVSALVLMA MVKRKDSRG 88 TCR-9 beta full- EPEVTQTPSH QVTQMGQEVI LRCVPISNHL YFYWYRQILG length QKVEFLVSFY NNEISEKSEI FDDQFSVERP DGSNFTLKIR STKLEDSAMY FCASTVQSPR TNEQFFGPGT RLTVL EDLKNVFPPE VAVFEPSEAE ISHTQKATLV CLATGFYPDH VELSWWVNGK EVHSGVSTDP QPLKEQPALN DSRYCLSSRL RVSATFWQNP RNHFRCQVQF YGLSENDEWT QDRAKPVTQI VSAEAWGRAD CGFTSESYQQ GVLSATILYE ILLGKATLYA VLVSALVLMA MVKRKDSRG 89 TCR-4 alpha variable QSVAQPEDQV NVAEGNPLIV KCTYSVSGNP YLFWYVQYPN domain RGLQFLLKYI TGDNLVKGSY GFEAEFNKSQ TSFHLKKPSA LVSDSALYFC AAPRDDKIIF GKGTRLHILP 90 TCR-4 alpha CDR1 VSGNPY 91 TCR-4 alpha CDR2 YITGDNLV 92 TCR-4 alpha CDR3 AAPRDDKII 751 TCR-4 alpha constant NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 93 TCR-4 alpha full- QSVAQPEDQV NVAEGNPLIV KCTYSVSGNP YLFWYVQYPN length RGLQFLLKYI TGDNLVKGSY GFEAEFNKSQTSFHLKKPSA LVSDSALYFC AAPRDDKIIF GKGTRLHILP NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 94 TCR-4 beta variable GVTQTPKFRV LKTGQSMTLL CAQDMNHEYM domain YWYRQDPGMG LRLIHYSVGE GTTAKGEVPD GYNVSRLKKQ NFLLGLESAA PSQTSVYFCA SSYLRTGGNE QFFGPGTRLT VL 66 TCR-4 beta CDR1 MNHEY 95 TCR-4 beta CDR2 SVGEGT 96 TCR-4 beta CDR3 ASSYLRTGGN EQF 11 TCR-4 beta constant EDLKNVFPPE VAVFEPSEAE ISHTQKATLV CLATGFYPDH domain VELSWWVNGK EVHSGVSTDP QPLKEQPALN DSRYCLSSRL RVSATFWQNP RNHFRCQVQF YGLSENDEWT QDRAKPVTQI VSAEAWGRAD CGFTSESYQQ GVLSATILYE ILLGKATLYA VLVSALVLMA MVKRKDSRG 97 TCR-4 beta full- GVTQTPKFRV LKTGQSMTLL CAQDMNHEYM length YWYRQDPGMG LRLIHYSVGE GTTAKGEVPD GYNVSRLKKQ NFLLGLESAA PSQTSVYFCA SSYLRTGGNE QFFGPGTRLT VL EDLKNVFPPE VAVFEPSEAE ISHTQKATLV CLATGFYPDH VELSWWVNGK EVHSGVSTDP QPLKEQPALN DSRYCLSSRL RVSATFWQNP RNHFRCQVQF YGLSENDEWT QDRAKPVTQI VSAEAWGRAD CGFTSESYQQ GVLSATILYE ILLGKATLYA VLVSALVLMA MVKRKDSRG 98 TCR-8 alpha variable QKVTQAQTEI SVVEKEDVTL DCVYETRDTT YYLFWYKQPP domain SGELVFLIRR NSFDEQNEIS GRYSWNFQKS TSSFNFTITA SQVVDSAVYF CALSGRGSGT YKYIFGTGTR LKVLA 99 TCR-8 alpha CDR1 TRDTTYY 100 TCR-8 alpha CDR2 RNSFDEQN 101 TCR-8 alpha CDR3 ALSGRGSGTY KYI 751 TCR-8 alpha constant NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 102 TCR-8 alpha full- QKVTQAQTEI SVVEKEDVTL DCVYETRDTT YYLFWYKQPP length SGELVFLIRR NSFDEQNEIS GRYSWNFQKS TSSFNFTITA SQVVDSAVYF CALSGRGSGT YKYIFGTGTR LKVLA NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 103 TCR-8 beta variable GVIQSPRHLI KEKRETATLK CYPIPRHDTV YWYQQGPGQD domain PQFLISFYEK MQSDKGSIPD RFSAQQFSDY HSELNMSSLE LGDSALYFCA SSAGTSYNEQ FFGPGTRLTV L 75 TCR-8 beta CDR1 PRHDT 76 TCR-8 beta CDR2 FYEKMQ 104 TCR-8 beta CDR3 ASSAGTSYNE QF 11 TCR-8 beta constant EDLKNVFPPE VAVFEPSEAE ISHTQKATLV CLATGFYPDH domain VELSWWVNGK EVHSGVSTDP QPLKEQPALN DSRYCLSSRL RVSATFWQNP RNHFRCQVQF YGLSENDEWT QDRAKPVTQI VSAEAWGRAD CGFTSESYQQ GVLSATILYE ILLGKATLYA VLVSALVLMA MVKRKDSRG 105 TCR-8 beta full- GVIQSPRHLI KEKRETATLK CYPIPRHDTV YWYQQGPGQD length PQFLISFYEK MQSDKGSIPD RFSAQQFSDY HSELNMSSLE LGDSALYFCA SSAGTSYNEQ FFGPGTRLIV L EDLKNVFPPE VAVFEPSEAE ISHTQKATLV CLATGFYPDH VELSWWVNGK EVHSGVSTDP QPLKEQPALN DSRYCLSSRL RVSATFWQNP RNHFRCQVQF YGLSENDEWT QDRAKPVTQI VSAEAWGRAD CGFTSESYQQ GVLSATILYE ILLGKATLYA VLVSALVLMA MVKRKDSRG 106 TCR-10 alpha variable QNIDQPTEMT ATEGAIVQIN CTYQTSGFNG LFWYQQHAGE domain APTFLSYNVL DGLEEKGRFS SFLSRSKGYS YLLLKELQMK DSASYLCAVP APYTGTASKL TFGTGTRLQV TL 107 TCR-10 alpha CDR1 TSGFNG 108 TCR-10 alpha CDR2 NVLDGL 109 TCR-10 alpha CDR3 AVPAPYTGTA SKLT 750 TCR-10 alpha constant DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 110 TCR-10 alpha full- QNIDQPTEMT ATEGAIVQIN CTYQTSGFNG LFWYQQHAGE length APTFLSYNVL DGLEEKGRFS SFLSRSKGYS YLLLKELQMK DSASYLCAVP APYTGTASKL TFGTGTRLQV TL DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 111 TCR-10 beta variable GVTQTPKFQV LKTGQSMTLQ CAQDMNHNSM domain YWYRQDPGMG LRLIYYSASE GTTDKGEVPN GYNVSRLNKR EFSLRLESAA PSQTSVYFCA SSEGYSTYNE QFFGPGTRLT VL 112 TCR-10 beta CDR1 MNHNS 113 TCR-10 beta CDR2 SASEGT 114 TCR-10 beta CDR3 ASSEGYSTYN EQF 11 TCR-10 beta constant EDLKNVFPPE VAVFEPSEAE ISHTQKATLV CLATGFYPDH domain VELSWWVNGK EVHSGVSTDP QPLKEQPALN DSRYCLSSRL RVSATFWQNP RNHFRCQVQF YGLSENDEWT QDRAKPVTQI VSAEAWGRAD CGFTSESYQQ GVLSATILYE ILLGKATLYA VLVSALVLMA MVKRKDSRG 115 TCR-10 beta full- GVTQTPKFQV LKTGQSMTLQ CAQDMNHNSM length YWYRQDPGMG LRLIYYSASE GTTDKGEVPN GYNVSRLNKR EFSLRLESAA PSQTSVYFCA SSEGYSTYNE QFFGPGTRLT VL EDLKNVFPPE VAVFEPSEAE ISHTQKATLV CLATGFYPDH VELSWWVNGK EVHSGVSTDP QPLKEQPALN DSRYCLSSRL RVSATFWQNP RNHFRCQVQF YGLSENDEWT QDRAKPVTQI VSAEAWGRAD CGFTSESYQQ GVLSATILYE ILLGKATLYA VLVSALVLMA MVKRKDSRG 116 TCR-12 alpha variable QLLEQSPQFL SIQEGENLTV YCNSSSVFSS LQWYRQEPGE domain GPVLLVTVVT GGEVKKLKRL TFQFGDARKD SSLHITAAQP GDTGLYLCAG GLSDSYDKVI FGPGTSLSVI P 117 TCR-12 alpha CDR1 SVFSS 118 TCR-12 alpha CDR2 VVTGGEV 119 TCR-12 alpha CDR3 AGGLSDSYDK VI 751 TCR-12 alpha constant NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 120 TCR-12 alpha full- QLLEQSPQFL SIQEGENLTV YCNSSSVFSS LQWYRQEPGE length GPVLLVTVVT GGEVKKLKRL TFQFGDARKD SSLHITAAQP GDTGLYLCAG GLSDSYDKVI FGPGTSLSVI P NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 121 TCR-12 beta variable EVTQTPKHLV MGMTNKKSLK CEQHMGHRAM YWYKQKAKKP domain PELMFVYSYE KLSINESVPS RFSPECPNSS LLNLHLHALQ PEDSALYLCA SSQGVGQEYG YTFGSGTRLT VV 58 TCR-12 beta CDR1 MGHRA 59 TCR-12 beta CDR2 YSYEKL 122 TCR-12 beta CDR3 ASSQGVGQEY GYT 32 TCR-12 beta constant EDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSW domain WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD F 123 TCR-12 beta full- EVTQTPKHLV MGMTNKKSLK CEQHMGHRAM YWYKQKAKKP length PELMFVYSYE KLSINESVPS RFSPECPNSS LLNLHLHALQ PEDSALYLCA SSQGVGQEYG YTFGSGTRLT VV EDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSW WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD F 124 TCR-11 alpha variable IQVEQSPPDL ILQEGANSTL RCNFSDSVNN LQWFHQNPWG domain QLINLFYIPS GTKQNGRLSA TTVATERYSL LYISSSQTTD SGVYFCAVNT GTASKLTFGT GTRLQVTL 125 TCR-11 alpha CDR1 DSVNN 126 TCR-11 alpha CDR2 IPSGT 127 TCR-11 alpha CDR3 AVNTGTASKL T 750 TCR-11 alpha constant DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 128 TCR-11 alpha full- IQVEQSPPDL ILQEGANSTL RCNFSDSVNN LQWFHQNPWG length QLINLFYIPS GTKQNGRLSA TTVATERYSL LYISSSQTTD SGVYFCAVNT GTASKLTFGT GTRLQVTL DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 129 TCR-11 beta variable GVTQTPKFQV LKTGQSMTLQ CAQDMNHNSM domain YWYRQDPGMG LRLIYYSASE GTTDKGEVPN GYNVSRLNKR EFSLRLESAA PSQTSVYFCA SSPRGQGRSY EQYFGPGTRL TVT 112 TCR-11 beta CDR1 MNHNS 113 TCR-11 beta CDR2 SASEGT 130 TCR-11 beta CDR3 ASSPRGQGRS YEQY 11 TCR-11 beta constant EDLKNVFPPE VAVFEPSEAE ISHTQKATLV CLATGFYPDH domain VELSWWVNGK EVHSGVSTDP QPLKEQPALN DSRYCLSSRL RVSATFWQNP RNHFRCQVQF YGLSENDEWT QDRAKPVTQI VSAEAWGRAD CGFTSESYQQ GVLSATILYE ILLGKATLYA VLVSALVLMA MVKRKDSRG 131 TCR-11 beta full- GVTQTPKFQV LKTGQSMTLQ CAQDMNHNSM length YWYRQDPGMG LRLIYYSASE GTTDKGEVPN GYNVSRLNKR EFSLRLESAA PSQTSVYFCA SSPRGQGRSY EQYFGPGTRL TVT EDLKNVFPPE VAVFEPSEAE ISHTQKATLV CLATGFYPDH VELSWWVNGK EVHSGVSTDP QPLKEQPALN DSRYCLSSRL RVSATFWQNP RNHFRCQVQF YGLSENDEWT QDRAKPVTQI VSAEAWGRAD CGFTSESYQQ GVLSATILYE ILLGKATLYA VLVSALVLMA MVKRKDSRG 132 TCR-2 alpha variable QKITQTQPGM FVQEKEAVTL DCTYDTSDQS YGLFWYKQPS domain SGEMIFLIYQ GSYDEQNATE GRYSLNFQKA RKSANLVISA SQLGDSAMYF CAMREGSDAG GTSYGKLTFG QGTILTVHP 24 TCR-2 alpha CDR1 TSDQSYG 25 TCR-2 alpha CDR2 QGSYDEQN 133 TCR-2 alpha CDR3 AMREGSDAGG TSYGKLT 751 TCR-2 alpha constant NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT domain DKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 134 TCR-2 alpha full- QKITQTQPGM FVQEKEAVTL DCTYDTSDQS YGLFWYKQPS length SGEMIFLIYQ GSYDEQNATE GRYSLNFQKA RKSANLVISA SQLGDSAMYF CAMREGSDAG GTSYGKLTFG QGTILIVHP NIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYIT DKIVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFP SPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTL RLWSS 135 TCR-2 beta variable GVIQSPRHLI KEKRETATLK CYPIPRHDTV YWYQQGPGQD domain PQFLISFYEK MQSDKGSIPD RFSAQQFSDY HSELNMSSLE LGDSALYFCA SSPSTGRLNT EAFFGQGTRL TVV 75 TCR-2 beta CDR1 PRHDT 76 TCR-2 beta CDR2 FYEKMQ 136 TCR-2 beta CDR3 ASSPSTGRLN TEAF 32 TCR-2 beta constant EDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSW domain WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD F 137 TCR-2 beta full- GVIQSPRHLI KEKRETATLK CYPIPRHDTV YWYQQGPGQD length PQFLISFYEK MQSDKGSIPD RFSAQQFSDY HSELNMSSLE LGDSALYFCA SSPSTGRLNT EAFFGQGTRL TVV EDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSW WVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQ NPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD CGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKD F

Re-Expression of TCRs

[0766] The human constant chain domains were exchanged by their murine counterparts with additional mutations in the transmembrane domain to enhance hydrophobicity. Both modifications are described in Jin et al., JCI Insight. 2018; 3(8):e99488.

[0767] In order to assess their functional characteristics, the identified TCRs were re-expressed in primary T cells of healthy donors using TCR mRNA electroporation technique. In brief, a T7 transcription was performed on DNA templates of the TCR alpha and beta chain nucleotide sequence. The resulting TCR mRNA was used for electroporation of pre-activated T cells for transient expression of the exogenous TCR which was validated in co-culture experiments.

Co-Culture Assays (Assessment of Functional Avidity, Specificity, and TCR-Motif)

[0768] The functional characteristics of the TCRs were assessed in co-culture experiments with T2 cells. CD8+ T cells were pre-stimulated and electroporated with TCR mRNA. At the day of co-culture, luciferase-transduced T2 cells were either loaded with peptide CT45-IP (SEQ ID NO: 138) at different concentrations (assessment of functional avidity, EC.sub.50), with sequence similar peptides (SEQ ID NO: 146-155) at a concentration of 10 μM or with alanine-substitution variants of the CT45-IP peptide (SEQ ID NO: 139-145) at a concentration range of 10-30 fold above the average EC.sub.50 of the respective TCR (TCR-motif determination). As control the irrelevant peptide NYESO1-001 (SEQ ID NO: 188) at a concentration of 10 μM and unloaded T2 cells were used. In brief, T2 cells were incubated for 2 hours with the respective amount of peptide and subsequently washed and harvested.

[0769] Cells electroporated with the model TCR 1G4-a95Ly as well as mock-electroporated T cells without exogenous TCR served as control. T cells and peptide-loaded T2 cells were seeded at a ratio of 1:1 and incubated for 24 h until supernatant harvest. Supernatants were subjected to an analysis for the presence of luciferase, released by apoptotic/necrotic T2 cells, killed by peptide-specific T cells. By adding specific substrate, the amount of luciferase present in the supernatant was determined by measuring the chemiluminescent signal in a microplate reader.

Test for Functionality in CD4+ T Cells

[0770] CD3+ T cells were stimulated and electroporated with TCR mRNA. After overnight incubation, the TCR-transfected T cells were co-cultivated at a 1:1 ratio with T2 cells loaded either with 10 μM CT45-IP (SEQ ID NO: 138) or 10 μM irrelevant NYESO1-001 (SEQ ID NO: 188) peptide. Directly after start of co-culture a cytokine secretion blocking reagent was added and incubated for 5 h at 37° C. Afterwards, the T cells were stained with fluorescently labeled antibodies for different surface markers such as CD4 and CD8. After fixation and permeabilization, the cells were stained for intracellular cytokines (TNF-α and IFN-γ) and analyzed on a flow cytometer (data not shown).

Lentiviral Co-Transduction of the Co-Stimulatory Molecule CD8

[0771] In order to engage CD4 T cells in the immune response after transduction with the MHC class I-restricted TCRs, a co-transduction with the co-stimulatory molecule CD8 was conducted. For that purpose, a lentiviral vector encoding the TCR chains as well as the CD8 molecule was used for transduction of pre-stimulated CD3+ T cells. The T cells were activated using plate-coated CD3 and anti-CD28 together with addition of IL-2 for 24 h. Pre-titrated concentrated lentiviral supernatant was added to the cells together with an adjuvant for transduction enhancement of lentivirus particles, e.g., Lentiboost® reagent (Sirion Biotech). The T cells were expanded over the course of 10 days using increasing volumes of media and decreasing concentration of IL-2 while transferring the cells consecutively into larger cell culture flasks. Transduction efficiency and resting state of T cells was checked prior to freezing of cells via flow cytometry. Subsequently, the effect of CD8 co-transfection on the killing efficiency against CT45-IP presenting tumor cells was analyzed in a live-cell monitoring killing assay (data not shown).

TCR Surface-Staining

[0772] A surface marker staining was performed with electroporated and pre-activated CD8+ T cells. To this end, anti-CD8, CD3 and/or mTCRB antibodies were used as well as TCR staining with fluorescently labeled Dextramers (dextramer backbone with conjugated CT45-IP-HLA-A*02 or irrelevant NYESO1-001-HLA-A*02). After 30 minutes cells were washed, fixed and subsequently analyzed by flow cytometry. Gates, to define dextramer-positive cells, were set according to the signal of cells stained with irrelevant peptide-MHC dextramer.

Live-Cell Monitoring Killing Assays

[0773] Proliferation of tumor cell lines expressing red fluorescent protein (RFP) was monitored using a live cell imaging system by quantifying red object counts over time. The cell lines NCIH1703 and A375 were co-cultivated with T cells expressing TCRs of the invention at an E:T ratio of 9:1, 3:1 or 1:1 or without T cells and monitored over a period of 48 h. As positive control, target cells were loaded with 10 μM of peptide CT45-IP. Decline of tumor cell line proliferation over time is an indicator for tumor cell killing.

Example 1: Functional Avidity

[0774] All fifteen identified CT45-IP-specific TCRs show high functional avidity as measured by peptide titration experiments which is expressed as half maximal killing capacity EC.sub.50 shown in FIG. 1. The measured EC.sub.50 values range from 0.15 nM to 59.5 nM. In particular the EC.sub.50 values are 7.78 nM (TCR-1); 4.69 nM (TCR-2); 1.02 nM (TCR-3); 1.31 nM (TCR-4); 1.32 nM (TCR-5); 3.25 nM (TCR-6); 0.48 nM (TCR-7); 6.52 nM (TCR-8); 0.15 nM (TCR-9); 8.75 nM (TCR-10); 59.50 nM (TCR-11); 47.47 nM (TCR-12); 11.38 nM (TCR-13); 17.69 nM (TCR-14); 18.60 nM (TCR-15).

Example 2: Specificity and TCR-Motif

[0775] The herein described TCRs were tested for their specificity profiles by testing their ability to recognize 10 CT45-IP sequence-similar peptides (SEQ ID NOs: 146-155). Loading with CT45-IP and CT45-IP sequence-similar peptides occurred at a very high concentration of 10 μM, to detect low signals. TCRs showed no binding to another peptide except CT45-IP. Minor binding signals were detected for TCR-9 and TCR-6 to peptide SP-05-0004 (SEQ ID NO: 149) as depicted in FIG. 2. The TCR characterization also involved the determination of the TCR binding motif to CT45-IP peptide as listed in Table 4. For this purpose, CT45-IP alanine exchange peptide variants (SEQ ID NO: 139-145) were tested in co-culture experiments with all fifteen TCRs. Alanine exchange peptides were tested at a concentration range of 10-30 fold above the average EC.sub.50 of the respective TCR. A relevant position is indicated by a number (referring to the position of the amino acid in the peptide), a non-relevant position is represented by a hyphen, and a position that was not tested is marked by an x.

TABLE-US-00004 TABLE 4 Consensus motif ID Consensus motif TCR-1 1x34567-x TCR-2 -x3(4)567-x TCR-3 -x34567-x TCR-4 -x34-678x TCR-5 -x3--67(8)x TCR-6 1x34-6--x TCR-7 1x-4-67-x TCR-8 -x---5-7-x TCR-9 1x34567(8)x TCR-10 -x--567-x TCR-11 1x3456-8x TCR-12 -x34-67-x TCR-13 1x34-67-x TCR-14 1x345---x TCR-15 1x34567-x

Example 3: CD4-Functionality

[0776] A test for functionality in CD4+ T cells was performed as described above. Among the fifteen tested TCRs, the inventors identified TCRs, which show functionality in CD8+ T cells as well as CD4+ T cells (data not shown).

Example 4: Surface Expression

[0777] The surface expression of the herein described TCRs was measured by CT45-IP-HLA-A2*02 dextramer staining and is shown in FIG. 3. The surface expression varied from 0.53% (TCR-11) to 55.5% (TCR-5) positive events after gating on CD3+ T cells after TCR mRNA electroporation. In particular, surface expression was 8.9% (TCR-1); 39.1% (TCR-2); 27.2% (TCR-3); 39.1% (TCR-4); 61.1% (TCR-5); 2.9% (TCR-6); 53.3% (TCR-7); 60.7% (TCR-8); 44.0% (TCR-9); 52.8% (TCR-10); 6.6% (TCR-11); 20.1% (TCR-12); 53.1% (TCR-13); 11.6% (TCR-14); 22.6% (TCR-15).

Example 5: Efficacy Against Tumor Cell Lines

[0778] Two tumor cell lines, the A375 with ˜30 copies per cell and the NCIH1703 with ˜150 copies per cell of CT45-IP were co-cultivated with CD8+ T cells expressing TCR-9, TCR-7 or Mock-TCR (FIG. 4). The efficacy of those TCRs for killing the two cell lines was assessed in a live cell monitoring experiment. While Mock-TCR does not lead to significant reduction of tumor cell proliferation, TCR-9 and TCR-7 efficiently kill the tumor cell lines loaded with CT45-IP peptide at an E:T ratio of 6:1 (TCR-9) or 1.8:1 (TCR-7) (NCIH1703 & A375), without additional peptide loading at an E:T ratio of 6:1 (TCR-9) or 1.8:1 (TCR-7) (NCIH1703 & A375), and at an E:T ratio of 2:1 (TCR-9) or 0.6:1 (TCR-7) and 0.6:1 (TCR-9) or 0.2:1 (TCR-7) (NICIH1703) as measured by the normalized red object count of <2.

Example 6: Safety Window

[0779] Co-culture assays were performed using T2 cells loaded with a titration series of peptide CT45-IP and SP-05-0004, respectively. The safety window was determined as EC.sub.50 SP 0004 divided by EC.sub.50 CT45-IP (data not shown).

Example 7: CT45-IP Presentation

[0780] The detection frequency of the CT45-IP antigenic peptide was analysed in primary and cultured tumor samples. A summary of the results is shown in Table 5. In the table, an expression of >0% is indicated as +, an expression of 10% is indicated as ++ and an expression of 30% is indicated as +++. The tumor entities in which presentation was detected are bile duct cancer (CCC), liver cancer (HCC), skin cancer (MEL, due to cell line identifications), lymph node cancer (NHL), non-small cell lung cancer (NSCLC), ovarian cancer (OC), esophageal cancer (OSCAR) and uterus cancer (UEC).

TABLE-US-00005 TABLE 5 Target presentation Target detection Entity frequency (%) CCC + HCC + MEL +++ NHL + NSCLC + OC ++ OSCAR ++ UEC +

Example 8: Efficacy of CD4+ T Cells Co-Transduced with a CT45-IP-Specific TCR and CD8 Against Tumor Cells

[0781] Tumor cells presenting CT45-IP were co-cultivated with CD4+ T cells or CD3+ T cells co-transduced with the herein described CT45-IP-specific TCRs and CD8. The killing efficacy of the T cells was assessed in a live cell monitoring experiment. Upon co-transduction with CT45-IP-specific TCR and CD8, efficient killing of the tumor cell lines as measured by the normalized red object count of <2 was observed (data not shown).

Items

[0782] 1. An antigen binding protein specifically binding to a CT45 antigenic peptide that is in a complex with a major histocompatibility complex (MHC) protein, wherein the CT45 antigenic peptide comprises or consists of the amino acid sequence of SEQ ID NO: 138 (KIFEMLEGV) and wherein the antigen binding protein comprises a first polypeptide comprising a variable domain V.sub.A comprising complementarity determining regions (CDR) CDRa1, CDRa2 and CDRa3 and a second polypeptide comprising a variable domain V.sub.B comprising CDRb1, CDRb2 and CDRb3, wherein [0783] 1) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 14, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 16, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 19, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 21, [0784] 2) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 24, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 133, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 75, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 136, [0785] 3) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 24, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 63, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 66, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 68, [0786] 4) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 90, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 92, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 66, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 96, [0787] 5) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 2, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 4, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 8, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 10, [0788] 6) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 53, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 55, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 58, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 60, [0789] 7) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 71, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 72, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 75, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 77, [0790] 8) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 99, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 101, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 75, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 104, [0791] 9) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 80, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 82, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 85, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 87, [0792] 10) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 107, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 109, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 112, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 114, [0793] 11) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 125, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 127, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 112, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 130, or [0794] 12) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 117, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 119, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 58, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 122, [0795] 13) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 24, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 35, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 38, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 40, [0796] 14) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 24, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 26, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 29, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 31, or [0797] 15) CDRa1 comprises or consists of the amino acid sequence of SEQ ID NO: 43, CDRa3 comprises or consists of the amino acid sequence of SEQ ID NO: 45, CDRb1 comprises or consists of the amino acid sequence of SEQ ID NO: 48, and CDRb3 comprises or consists of the amino acid sequence of SEQ ID NO: 50, [0798] wherein the antigen binding protein comprises said CDRa1, CDRa3, CDRb1 and CDRb3 sequence(s) with not more than one, two or three amino acid mutations, wherein each of CDRa1, CDRa3, CDRb1 and/or CDRb3 may comprise one, two or three amino acid mutations. [0799] 2. The antigen binding protein of item 1, wherein [0800] 1) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 15, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 20, [0801] 2) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 25, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 76, [0802] 3) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 25, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 67, [0803] 4) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 91, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 95, [0804] 5) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 3, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 9, [0805] 6) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 54, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 59, [0806] 7) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 15, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 76, [0807] 8) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 100, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 76, [0808] 9) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 81, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 86, [0809] 10) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 108, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 113, [0810] 11) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 126, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 113, or [0811] 12) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 118, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 59, [0812] 13) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 25, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 39, [0813] 14) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 25, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 30, or [0814] 15) CDRa2 comprises or consists of the amino acid sequence of SEQ ID NO: 44, and CDRb2 comprises or consists of the amino acid sequence of SEQ ID NO: 49, [0815] wherein the antigen binding protein comprises said CDRa2 and CDRb2 sequence(s) with not more than one, two, three or four amino acid mutations, wherein each of CDRa2 and/or CDRb2 may comprise one, two, three or four amino acid mutations. [0816] 3. The antigen binding protein of item 1 or 2, wherein the antigen binding protein specifically binds to a complex of the CT45 antigenic peptide and an MHC protein. [0817] 4. The antigen binding protein of any one of items 1 to 3, wherein the MHC protein is an HLA protein, more particularly HLA-A, even more particularly HLA-A*02. [0818] 5. The antigen binding protein of any one of items 1 to 4, wherein the EC.sub.50 of CT45-IP for inducing killing of CT45-IP:MHC complex presenting cells by T cells expressing the antigen binding protein is less than about 60 nM, less than about 50 nM, less than about 30 nM, less than about 25 nM, less than about 20 nM, less than about 15 nM, less than about 10 nM, less than about 5 nM, less than about 2.5 nM, less than about 1.5 nM or less than about 1 nM. [0819] 6. The antigen binding protein of any one of items 1 to 5, wherein the antigen binding protein specifically binds to a functional epitope comprising or consisting of 2, 3 or 4 amino acid positions selected from the group consisting of positions 3, 4, 5, 6 and 7 of SEQ ID NO: 138. [0820] 7. The antigen binding protein of any one of items 1 to 6, wherein the antigen binding protein does not significantly bind to at least 1, at least 2, at least 3, at least 4, at least 5, or all similar peptides selected from the group consisting of SEQ ID NO: 146 (SP-05-0001), SEQ ID NO: 147 (SP-05-0002), SEQ ID NO: 148 (SP-05-0003), SEQ ID NO: 149 (SP-05-0004), SEQ ID NO: 150 (SP-05-0005), SEQ ID NO: 151 (SP-05-0006), SEQ ID NO: 152 (SP-05-0007), SEQ ID NO: 153 (SP-05-0008), SEQ ID NO: 154 (SP-05-0009) and SEQ ID NO: 155 (SP-05-0010), preferably from the group consisting of SEQ ID NO: 146 (SP-05-0001), SEQ ID NO: 147 (SP-05-0002), SEQ ID NO: 148 (SP-05-0003), SEQ ID NO: 150 (SP-05-0005), SEQ ID NO: 151 (SP-05-0006), SEQ ID NO: 152 (SP-05-0007), SEQ ID NO: 153 (SP-05-0008), SEQ ID NO: 154 (SP-05-0009) and SEQ ID NO: 155 (SP-05-0010), more preferably from the group consisting of SEQ ID NO: 147 (SP-05-0002), SEQ ID NO: 151 (SP-05-0006), SEQ ID NO: 152 (SP-05-0007) and SEQ ID NO: 155 (SP-05-0010). [0821] 8. The antigen binding protein of any one of items 1 to 7, wherein the first and the second polypeptide are comprised on two polypeptide chains, preferably wherein V.sub.A is comprised in a first polypeptide chain and V.sub.B is comprised in a second polypeptide chain. [0822] 9. The antigen binding protein of any one of items 1 to 7, wherein the antigen binding protein is a single chain antigen binding protein, preferably a single chain TCR, or a single chain bispecific antigen binding protein, preferably a single chain bispecific TCR. [0823] 10. The antigen binding protein of any one of items 1 to 9, wherein the antigen binding protein is monovalent or multivalent, e.g. tetra-, tri- or bivalent. [0824] 11. The antigen binding protein of any one of items 1 to 10, wherein the antigen binding protein is monospecific or multispecific, e.g. tetra-, tri- or bispecific. [0825] 12. The antigen binding protein of any one of items 1 to 11, wherein the antigen binding protein is a soluble protein. [0826] 13. The antigen binding protein of any one of items 1 to 12, wherein the antigen binding protein is a TCR. [0827] 14. The antigen binding protein of item 13, wherein the TCR is selected from the group consisting of an α/β TCR, a γ/δ TCR, a single chain TCR, a membrane-bound TCR, a soluble TCR, a monovalent, bivalent or multivalent TCR, a monospecific, bispecific or multispecific TCR, a functional fragment of a TCR, and a fusion protein or chimeric protein comprising a functional fragment of a TCR. [0828] 15. The antigen binding protein of item 13, wherein the TCR is an α/β TCR or a γ/δ TCR, preferably an α/β TCR. [0829] 16. The antigen binding protein of any one of items 1 to 15, further comprising one or more of the following: [0830] (i) one or more further antigen binding sites; [0831] (ii) a transmembrane domain, optionally including a cytoplasmic signaling region; [0832] (iii) a diagnostic agent; [0833] (iv) a therapeutic agent. [0834] 17. The antigen binding protein of item 16, wherein the one or more further antigen binding sites comprise an antibody-derived antigen binding site, preferably comprising or consisting of V.sub.L and V.sub.H. [0835] 18. The antigen binding protein of any one of items 1 to 17, wherein V.sub.A comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 13, 132, 62, 89, 1, 52, 70, 98, 79, 106, 124, 116, 34, 23, and 42, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 13, 132, 62, 89, 1, 52, 70, 98, 79, 106, 124, 116, 34, 23, or 42 and comprising the CDRa1, CDRa2 and CDRa3 according to item 1 or 2; and wherein V.sub.B comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 18, 135, 65, 94, 7, 57, 74, 103, 84, 111, 129, 121, 37, 28 and 47 or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 18, 135, 65, 94, 7, 57, 74, 103, 84, 111, 129, 121, 37, 28 or 47 and comprising the CDRb1, CDRb2 and CDRb3 according to item 1 or 2, wherein the CDRa1, CDRa2, CDRa3, CDRb1, CDRb2 and/or CDRb3 sequences may comprise one, two or three amino acid mutations, preferably amino acid substitutions. [0836] 19. The antigen binding protein of any one of items 1 to 18, wherein V.sub.A and V.sub.B are TCR variable domains, preferably TCR alpha, beta, gamma or delta variable domains, more preferably wherein V.sub.A is a TCR alpha or gamma, preferably alpha, variable domain, and V.sub.B is a TCR beta or delta, preferably beta, variable domain. [0837] 20. The antigen binding protein of any one of items 1 to 19, further comprising a constant domain, wherein the constant domain comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 5, 750, 751, 156, 11, 32, and 157, preferably selected from the group consisting of SEQ ID NO: 5, 750, 751, 11 and 32, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 5, 750, 751, 156, 11, 32 or 157. [0838] 21. The antigen binding protein of any one of items 1 to 20, wherein the first polypeptide is a TCR alpha chain and the second polypeptide is a TCR beta chain or the first polypeptide is a TCR gamma chain and the second polypeptide is a TCR delta chain preferably wherein the first polypeptide is a TCR alpha chain and the second polypeptide is a TCR beta chain. [0839] 22. The antigen binding protein of any one of items 1 to 21, wherein the first polypeptide comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 17, 134, 64, 93, 6, 56, 73, 102, 83, 110, 128, 120, 36, 27, 46 and 158-172, preferably selected from the group consisting of SEQ ID NO: 17, 134, 64, 93, 6, 56, 73, 102, 83, 110, 128, 120, 36, 27, 46 or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 17, 134, 64, 93, 6, 56, 73, 102, 83, 110, 128, 120, 36, 27, 46 or 158-172, and the second polypeptide comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 22, 137, 69, 97, 12, 61, 78, 105, 88, 115, 131, 123, 41, 33, 51 and 173-187, preferably selected from the group consisting of SEQ ID NO: 22, 137, 69, 97, 12, 61, 78, 105, 88, 115, 131, 123, 41, 33, 51, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 22, 137, 69, 97, 12, 61, 78, 105, 88, 115, 131, 123, 41, 33, 51 or 173-187. [0840] 23. The antigen binding protein of any one of items 1 to 22, wherein the antigen binding protein specifically binds to a functional epitope comprising or consisting of [0841] i. amino acid positions 1, 3 and 4 of SEQ ID NO: 138, preferably amino acid positions 1, 3, 4 and 5, or 1, 3, 4 and 6 or 1, 3, 4, 5 and 6 or 1, 3, 4, 5, 6 and 7 of SEQ ID NO: 138; [0842] ii. amino acid positions 4, 6 and 7 of SEQ ID NO: 138, preferably amino acid positions 1, 4, 6 and 7, or 3, 4, 6 and 7 or 1, 3, 4, 6 and 7 of SEQ ID NO: 138; or [0843] iii. amino acid positions 5 and 7 of SEQ ID NO: 138, preferably amino acid positions 5, 6 and 7, or 3, 4, 5, 6 and 7 of SEQ ID NO: 138. [0844] 24. The antigen binding protein of any one of items 1 to 23, wherein the antigen binding protein does not significantly bind to the similar peptides of the group consisting of [0845] i. SEQ ID NO: 146 (SP-05-0001), SEQ ID NO: 147 (SP-05-0002), SEQ ID NO: 148 (SP-05-0003), SEQ ID NO: 149 (SP-05-0004), SEQ ID NO: 150 (SP-05-0005), SEQ ID NO: 151 (SP-05-0006), SEQ ID NO: 152 (SP-05-0007), SEQ ID NO: 153 (SP-05-0008), SEQ ID NO: 154 (SP-05-0009) and SEQ ID NO: 155 (SP-05-0010); or [0846] ii. SEQ ID NO: 146 (SP-05-0001), SEQ ID NO: 147 (SP-05-0002), SEQ ID NO: 148 (SP-05-0003), SEQ ID NO: 150 (SP-05-0005), SEQ ID NO: 151 (SP-05-0006), SEQ ID NO: 152 (SP-05-0007), SEQ ID NO: 153 (SP-05-0008), SEQ ID NO: 154 (SP-05-0009) and SEQ ID NO: 155 (SP-05-0010). [0847] 25. The antigen binding protein of any one of items 1 to 24, wherein the antigen binding protein has a mean expression of at least 5%, at least 10%, at least 20%, at least 30%, or at least 40%. [0848] 26. The antigen binding protein of any one of items 1 to 25, wherein V.sub.A comprises a V segment encoded by TRAV14, in particular TRAV14/DV4, and a CDRa1 according to SEQ ID NO: 24 and a CDRa2 according to SEQ ID NO: 25. [0849] 27. The antigen binding protein of any one of items 1 to 26, wherein V.sub.B comprises [0850] i. a V segment encoded by TRBV13, and a CDRb1 according to SEQ ID NO: 75 and a CDRb2 according to SEQ ID NO: 76; [0851] ii. a V region encoded by TRBV4-1, and a CDRb1 according to SEQ ID NO: 58 and a CDRb2 according to SEQ ID NO: 59, or [0852] iii. a V region encoded by TRBV6-1, and a CDRb1 according to SEQ ID NO: 112 and a CDRb2 according to SEQ ID NO: 113. [0853] 28. The antigen binding protein of any one of items 1 to 27, wherein the antigen binding protein is capable of activating [0854] a CD4+ T cell, in particular a CD4+CD8− T cell, and/or [0855] a CD8+ T cell, in particular a CD8+CD4-T cell, and wherein the antigen binding protein is preferably a TCR, more preferably an α/β TCR or γ/δ TCR. [0856] 29. A nucleic acid or nucleic acids comprising one or more sequences encoding the antigen binding protein of any one of items 1 to 28. [0857] 30. A vector or a collection of vectors comprising the nucleic acid(s) of item 29. [0858] 31. A host cell comprising the antigen binding protein of any one of items 1 to 28, or the nucleic acid(s) of item 29, or the vector or collection of vectors of item 30. [0859] 32. The host cell of item 31, wherein the host cell is [0860] a lymphocyte, preferably a T cell, a T cell progenitor or an NK cell, more preferably a CD4 or CD8 positive T cell; or [0861] a cell for recombinant expression, such as a Chinese Hamster Ovary (CHO) cell or a yeast cell. [0862] 33. A pharmaceutical composition comprising the antigen binding protein of any one of items 1 to 28, the nucleic acid(s) of item 29, the vector or collection of vectors of item 30, or the host cell of item 31 or 32 and optionally a pharmaceutically acceptable carrier. [0863] 34. The antigen binding protein of any one of items 1 to 28, the nucleic acid(s) of item 29, the vector or collection of vectors of item 30, the host cell of item 31 or 32, or the pharmaceutical composition of item 33 for use in medicine. [0864] 35. The antigen binding protein of any one of items 1 to 28, the nucleic acid(s) of item 29, the vector or collection of vectors of item 30, the host cell of item 31 or 32, or the pharmaceutical composition of item 33 for use in a method of treatment and/or diagnosis of a proliferative disease, in particular cancer. [0865] 36. A method of treatment of a proliferative disease, in particular cancer, comprising administering to a subject in need thereof a therapeutically effective amount of the antigen binding protein of any one of items 1 to 28, the nucleic acid(s) of item 29, the vector or collection of vectors of item 30, the host cell of item 31 or 32, or the pharmaceutical composition of item 33. [0866] 37. The antigen binding protein, nucleic acid(s), vector or collection of vectors, host cell or pharmaceutical composition for use of item 35 or the method of treatment of item 36, wherein the cancer is a CT45 expressing cancer, more particularly selected from the group of cancers consisting of lung cancer, NSCLC, gall bladder cancer, bile duct cancer, lymph node cancer, ovarian cancer, esophageal cancer, liver cancer, uterus cancer and melanoma. [0867] 38. The antigen binding protein, nucleic acid(s), vector or collection of vectors, host cell or pharmaceutical composition for use of item 35 or 37 or the method of treatment of item 36 or 37, wherein the method of treatment comprises immune therapy, in particular adoptive autologous or heterologous T-cell therapy. [0868] 39. The antigen binding protein, nucleic acid(s), vector or collection of vectors, host cell or pharmaceutical composition for use of any one of items 35, 37 or 38 or the method of treatment of any one of items 36 to 38, wherein the antigen binding protein is a TCR. [0869] 40. The antigen binding protein, nucleic acid(s), vector or collection of vectors, host cell or pharmaceutical composition for use of any one of items 35 or 37 to 39 or the method of treatment of any one of items 36 to 39, wherein the antigen binding protein is expressed on the surface of a host cell. [0870] 41. The antigen binding protein, nucleic acid(s), vector or collection of vectors, host cell or pharmaceutical composition for use of any one of items 35 or 37 to 40 or the method of treatment of any one of items 36 to 40, wherein the method of treatment comprises administration of a host cell expressing the antigen binding protein, wherein the host cell is a T cell, a T cell progenitor or an NK cell, preferably a T cell. [0871] 42. The antigen binding protein, nucleic acid(s), vector or collection of vectors, host cell or pharmaceutical composition for use of item 40 or the method of treatment of item 40, wherein the host cell, preferably the T cell, T cell progenitor or NK cell, more preferably the T cell, is autologous. [0872] 43. The antigen binding protein, nucleic acid(s), vector or collection of vectors, host cell or pharmaceutical composition for use of item 40 or the method of treatment of item 40, wherein the host cell, preferably the T cell, T cell progenitor or NK cell, more preferably the T cell, is allogeneic. [0873] 44. The antigen binding protein, nucleic acid(s), vector or collection of vectors, host cell or pharmaceutical composition for use of any one of items 35 or 37 to 43 or the method of treatment of any one of items 36 to 43, wherein the antigen binding protein is conjugated to a therapeutically active agent. [0874] 45. The antigen binding protein, nucleic acid(s), vector or collection of vectors, host cell or pharmaceutical composition for use of any one of items 44 or the method of treatment of any one of items 44, wherein the therapeutically active agent is selected from the group consisting of a radionuclide, a chemotherapeutic agent and a toxin. [0875] 46. The antigen binding protein, nucleic acid(s), vector or collection of vectors, host cell or pharmaceutical composition for use of any one of items 35 or 37 to 45 or the method of treatment of any one of items 36 to 45, wherein the method of treatment further comprises administering at least one chemotherapeutic agent to the subject in need of treatment. [0876] 47. The antigen binding protein, nucleic acid(s), vector or collection of vectors, host cell or pharmaceutical composition for use of any one of items 35 or 37 to 46 or the method of treatment of any one of items 36 to 46, wherein the method of treatment further comprises administering radiation therapy to the subject in need of treatment. [0877] 48. A method of treating cancer in a subject in need thereof, comprising: [0878] a) isolating a cell from said subject; [0879] b) transforming the cell with a vector or collection of vectors encoding the antigen binding protein of any one of items 1 to 28 to produce a transformed cell; [0880] c) expanding the transformed cell to produce a plurality of transformed cells; and [0881] d) administering the plurality of transformed cells to said subject. [0882] 49. A method of treating cancer in a subject in need thereof, comprising: [0883] a) isolating a cell from a healthy donor; [0884] b) transforming the cell with a vector or collection of vectors encoding the antigen binding protein of any one of items 1 to 28 to produce a transformed cell; [0885] c) expanding the transformed cell to produce a plurality of transformed cells; and [0886] d) administering the plurality of transformed cells to said subject. [0887] 50. The method of item 48 or 49, wherein the transformed cell is a lymphocyte, preferably an NK cell or T cell or T cell progenitor, more preferably a T cell. [0888] 51. Use of the antigen binding protein of any one of items 1 to 28 for the manufacture of a medicament for the treatment of a proliferative disease. [0889] 52. An in-vitro method of detecting cancer, in particular cancer expressing CT45, in a biological sample comprising: [0890] a) contacting the biological sample with the antigen binding protein of any one of items 1 to 28, and [0891] b) detecting binding of the antigen binding protein to the biological sample. [0892] 53. A method of producing the antigen binding protein according to any one of items 1 to 28, comprising [0893] a) providing a host cell, [0894] b) providing a genetic construct comprising a nucleic acid or nucleic acids encoding the antigen binding protein of any of items 1 to 28, [0895] c) introducing the genetic construct into the host cell, and [0896] d) expressing the genetic construct by the host cell. [0897] 54. The method of item 53, further comprising the isolation and purification of the antigen binding protein from the host cell and, optionally, reconstitution of the antigen binding protein in a T cell. [0898] 55. The method of item 53 or 54, further comprising cell surface presentation of said antigen binding protein. [0899] 56. The method of any one of items 53 to 55, wherein the genetic construct is an expression construct comprising a promoter sequence operably linked to the nucleic acid encoding the antigen binding protein. [0900] 57. The method according to any one of items 53 to 56, wherein the genetic construct is introduced into the host cell by retroviral transfection.