RU2849866C1 - T-cell receptors, restricted by hla class i, against ras with mutation g12d - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, in particular to a T-cell receptor (TCR). Said TCR contains CDR1 chain α, containing SEQ ID NO: 1, CDR2 chain α, containing SEQ ID NO: 2, CDR3 chain α, containing SEQ ID NO: 3, CDR1 chain β, containing SEQ ID NO: 4, CDR2 chain β, containing SEQ ID NO: 5, and CDR3 chain β, containing SEQ ID NO: 6, where said TCR has antigen specificity for the amino acid sequence of the mutant human RAS with the replacement of the glycine at position 12 with aspartic acid. Invention also relates to a polypeptide and protein containing said CDR, nucleic acid, recombinant expression vector, a host cell, a cell population, a pharmaceutical composition for treating cancer, a method of treating cancer and a method of inducing an immune response to cancer.

EFFECT: present invention provides a protein with antigen specificity with respect to the amino acid sequence of mutant human RAS with the replacement of glycine in position 12 with aspartic acid.

29 cl, 5 dwg, 7 tbl, 9 ex

Description

Cross-reference to a related application

[0001] This patent application claims priority to U.S. Patent Application No. 62/975,544, filed February 12, 2020, which is incorporated herein by reference in its entirety.

Statement regarding federally funded research or development

[0002] This invention was developed with the support of the government under project number ZIABC010984 by the National Institutes of Health, National Cancer Institute. The state has certain rights in this invention.

Incorporation by reference of material presented in electronic form

[0003] The machine-readable listing of nucleotide/amino acid sequences filed herewith and identified as follows is incorporated by reference in its entirety herein: one (text) file, 114961 bytes ASCII, named "751506_ST25.txt", dated January 29, 2021.

Prior art

[0004] Some cancers may have very limited treatment options, particularly when the cancer becomes metastatic and inoperable. Despite advances in treatments such as surgery, chemotherapy, and radiation therapy, the prognosis for many cancers, such as pancreatic cancer, colorectal cancer, lung cancer, endometrial cancer, ovarian cancer, and prostate cancer, can be poor. Accordingly, there is an unmet need for additional cancer treatments.

Brief summary of the invention

[0005] According to one embodiment of the invention, there is provided an isolated or purified T cell receptor (TCR) comprising the amino acid sequences of (a) SEQ ID NO: 1-3, (b) SEQ ID NO: 4-6, or (c) SEQ ID NO: 1-6, wherein the TCR has antigen specificity for the amino acid sequence of a mutant human RAS with a glycine substitution at position 12 with aspartic acid, presented by a class I human leukocyte antigen (HLA) molecule, wherein the amino acid sequence of the mutant human RAS is the amino acid sequence of a mutant human Kirsten rat sarcoma viral oncogene homolog (KRAS), a mutant human Harvey rat sarcoma viral oncogene homolog (HRAS), or a mutant human Harvey rat sarcoma viral oncogene homolog) or a mutant human homolog of the neuroblastoma rat sarcoma viral oncogene RAS (NRAS - from English Neuroblastoma rat sarcoma viral oncogene homolog), and where position 12 is determined taking into account the wild-type human KRAS protein, wild-type human HRAS protein, or wild-type human NRAS protein, respectively.

[0006] According to another embodiment of the invention, there is provided an isolated or purified polypeptide comprising a functional portion of a TCR of the invention, wherein the functional portion comprises the following amino acid sequences: (a) all of SEQ ID NOs: 1-3, (b) all of SEQ ID NOs: 4-6, or (c) all of SEQ ID NOs: 1-6.

[0007] According to another embodiment of the invention, there is provided an isolated or purified protein comprising a first polypeptide chain comprising the amino acid sequences of SEQ ID NO: 1-3 and a second polypeptide chain comprising the amino acid sequences of SEQ ID NO: 4-6.

[0008] According to embodiments of the invention, nucleic acids, recombinant expression vectors, host cells, cell populations and pharmaceutical compositions related to the TCRs, polypeptides and proteins of the invention are further provided.

[0009] According to an embodiment of the invention, there is provided an isolated or purified nucleic acid comprising, from 5' to 3', a first nucleic acid sequence and a second nucleic acid sequence, wherein the first and second nucleotide sequences, respectively, encode the amino acid sequences of SEQ ID NO: 7 and 8; 51 and 8; 7 and 52; 51 and 52; 8 and 7; 8 and 51; 52 and 7; 52 and 51; 21 and 22; 53 and 22; 21 and 54; 53 and 54; 22 and 21; 22 and 53; 54 and 21; 54 and 53; 23 and 24; 55 and 24; 23 and 56; 55 and 56; 24 and 23; 24 and 55; 56 and 23; 56 and 55; 32 and 33; 33 and 32; 59 and 60; 60 and 59; 34 and 35; 35 and 34; 61 and 62; 62 and 61; 36 and 37; 37 and 36; 63 and 64; 64 and 63; 40 and 41; 57 and 41; 40 and 58; 57 and 58; 41 and 40; 41 and 57; 58 and 40; 58 and 57; 42 and 43; 43 and 42; 65 and 66; or 66 and 65.

[0010] Methods for detecting the presence of cancer in a mammal, methods for treating or preventing cancer in a mammal, methods for inducing an immune response to cancer in a mammal, methods for producing a host cell expressing a TCR that has antigen specificity for a peptide (SEQ ID NO: 29), and methods for producing TCRs, polypeptides and proteins of the invention are further provided according to embodiments of the invention.

[0011] Additional embodiments are as described herein.

Brief description of several representations of graphic material(s)

[0012] Fig. 1A-1B: TIL screening for KRAS G12D reactivity. Fig. 1A is a graph showing the ELISPOT measurement of IFN-γ secretion (number of spots per 3e4 cells). Fig. 1B is a graph showing the results of flow cytometry analysis of 4-1 BB and OX40 expression (% 4-1 BB+/OX40+) measured after co-culture of effector cells with target cells. The effector cells were TILs from Patient 4373 tumor fragments F4, F5, F6, F8, F9, and F10. Target cells (autologous DCs) were electroporated with mRNA using a tandem minigene (TMG) encoding 12 RAS mutations (G12D- G12V- G12C- G12A- G12S-G13D- G13R- G13V-Q61R- Q61L- Q61K-Q61H) (SEQ ID NO: 49) (open circles) or wild-type (WT) RAS epitope sequences (WT G12+G13+Q61) (SEQ ID NO: 48) (filled circles); autologous DCs loaded with the long peptide G12 WT (LP) (SEQ ID NO: 27) (filled squares); G12D Mut LP (SEQ ID NO: 26) (open squares); or the minimal epitope (ME) A11/A02 - a mixture of equal concentrations of three peptide sequences: KLVWGADGV (SEQ ID NO: 50), WGADGVGK (SEQ ID NO: 28), WVGADGVGK (SEQ ID NO: 29) (filled triangles). As negative controls, autologous DC cells were cultured alone (TIL only) (diamonds) or co-cultured with dimethyl sulfoxide (DMSO) (open triangles). As a positive control, TIL grew in the presence of the material - magnetic beads with anti-CD3/anti-CD28 antibodies (ThermoFisher) (asterisks).

[0013] Fig. 2 is a graph showing the percentage of cells expressing 4-1 BB and OX40 after co-culture of effector cells with target cells. The effector cells were autologous PBLs from Patient 4373 transduced with (i) one of two TCR sequences (TCR1 or TCR2) obtained by single-cell sequencing from reactive TILs (shown in Fig. 1) suspected to be G12D RAS-reactive, or (ii) HLA-A11-restricted, murine anti-KRAS G12D TCR (positive control) (mTCR), or (iii) untransduced PBLs (-). Target cells were: COS HLA-A2 cell line (open circles), COS HLA-A2-G12D cell line (filled circles), COS HLA-A11 cell line (open triangles), or COS HLA-A11-G12D cell line (filled triangles), or T cells only (no target cells) (-).

[0014] Fig. 3A-3B: 4373 TCR-transduced PBL tested for KRAS G12D reactivity. Fig. 3A is a graph showing the ELISPOT measurement of IFN-γ secretion (number of spots per 3e4 cells). Fig. 3B is a graph showing the results of a flow cytometry analysis of 4-1 BB and OX40 expression (% 4-1 BB+/OX40+) of CD8 sorted cells measured after co-culture of effector cells with target cells. The effector cells were autologous CD8+ PBL from Patient 4373 transduced with a retroviral expression vector encoding (i) 4373 TCRs 1 and 2, which are suspected to be G12D RAS-reactive (with human variable regions); (ii) the HLA-A11-restricted, murine anti-KRAS G12D TCR of Example 1; or (iii) green fluorescent protein (GFP) (control). Cells transduced with empty vector served as an additional control (Mock Td). Target cells were autologous dendritic cells (DCs) transfected with mRNA for the full-length (FL) gene, which was either the KRAS WT gene (open circles) or the FL gene with the KRAS G12D mutation (filled circles); autologous DCs loaded with a peptide that was WT KRAS LP (open triangles) or G12D Mut KRAS LP (filled triangles), or loaded with a minimal KRAS epitope (ME): ME A02 WT (open diamonds); ME A02 G12D (filled diamonds); a mixture of ME HLA-A11 WT (open squares); or a mixture of ME HLA-A11 G12D (filled squares). As controls, transduced cells were cultured alone (T cells only) (asterisks) or cultured with DMSO (stars) or the material—magnetic beads with anti-CD28/anti-CD3 antibodies (hexagons).

[0015] Fig. 4A-4C: TCR avidity testing against autologous DCs loaded with ME titration: Fig. 4A is a graph showing the results of IFN-γ secretion ELISPOT (number of spots per 3e4 cells). Fig. 4B and 4C are graphs showing the results of flow cytometry analysis of 4-1 BB and OX40 expression (% 4-1 BB+/OX40+) (4B) in CD8 sorted and in CD4 sorted mTCR-positive PBL (4C), measured after co-culture of effector cells with target cells. Effector cells were autologous PBL from Patient 4373 transduced with a retroviral expression vector encoding the G12D RAS-reactive 4373 TCR of Example 2 (with human variable regions). Target cells were autologous DC from Patient 4373 loaded with the following peptides at the concentrations shown: mutant 10-amino acid residue minimal epitope peptide (VWGADGVGK) (SEQ ID NO: 29) (open circles) or WT 10-amino acid residue minimal epitope peptide (SEQ ID NO: 31) (filled circles).

[0016] Fig. 5A-5C: TCR avidity testing against the autologous COS-A11 cell line loaded with ME titration: Fig. 5A is a graph showing the results of IFN-γ secretion ELISPOT (number of spots per 3e4 cells). Fig. 5B and 5C are graphs showing the results of flow cytometry analysis of 4-1 BB and OX40 expression (% 4-1 BB+/OX40+) in CD8 sorted (5B) and CD4 sorted mTCR-positive PBL (5C), measured after co-culture of effector cells with target cells. Effector cells were autologous PBL from Patient 4373 transduced with a retroviral expression vector encoding the G12D RAS-reactive 4373 TCR of Example 2 (with human variable regions). Target cells were autologous DC from Patient 4373 loaded with the following peptides at the concentrations shown: mutant minimal peptide - 10-amino acid residue epitope (SEQ ID NO: 29) (open circles) or minimal peptide - WT epitope with 10 amino acid residues (SEQ ID NO: 31) (filled circles).

Detailed description of the invention

[0017] The RAS family proteins belong to a large family of small GTPases. Without being bound by a specific theory or mechanism, it is believed that, when mutant, RAS proteins may participate in signaling early in the oncogenesis of many human cancers. A single amino acid substitution can activate the protein. A mutant RAS protein product can be constitutively activated. Mutant RAS proteins can be expressed in any variety of human cancers, such as, for example, pancreatic cancer (e.g., pancreatic carcinoma), colorectal cancer, lung cancer (e.g., lung adenocarcinoma), endometrial cancer, ovarian cancer (e.g., epithelial ovarian cancer), and prostate cancer. Human RAS family proteins include KRAS, HRAS, and NRAS.

[0018] KRAS is also called KRas GTPase, Kirsten rat sarcoma viral oncogene V-Ki-Ras2, or KRAS2. There are two variants of the KRAS transcript: KRAS variant A and KRAS variant B. The wild-type (WT) KRAS variant A has the amino acid sequence of SEQ ID NO: 9. The WT KRAS variant B has the amino acid sequence of SEQ ID NO: 10. Hereinafter, references to "KRAS" (mutant or non-mutant (WT)) refer to both variant A and variant B unless specifically stated otherwise. When activated, mutant KRAS binds to guanosine 5'-triphosphate (GTP) and converts GTP to guanosine 5'-diphosphate (GDP).

[0019] HRAS is another member of the RAS protein family. HRAS is also known as Harvey rat sarcoma viral oncoprotein, Harvey rat sarcoma viral oncogene homolog V-Ha-Ras, or small GTP-binding protein of the Ras family H-Ras. HRAS WT has the amino acid sequence of SEQ ID NO: 11.

[0020] NRAS is another member of the RAS protein family. NRAS is also known as NRas GTPase, neuroblastoma viral oncogene RAS homolog V-Ras, or NRAS1. NRAS WT has the amino acid sequence SEQ ID NO: 12.

[0021] According to one embodiment, an isolated or purified TCR is provided, wherein the TCR has antigen specificity for the amino acid sequence of a mutant human RAS with a glycine substitution at position 12 with aspartic acid, wherein the amino acid sequence of the mutant human RAS is the amino acid sequence of a mutant human KRAS, a mutant human HRAS, or a mutant human NRAS, and wherein position 12 is determined based on a human KRAS WT, human HRAS WT, or human NRAS WT protein, respectively. Hereinafter, references to "TCR" also refer to functional portions and functional variants of a TCR, unless otherwise stated.

[0022] The amino acid sequence of the mutant human RAS may be the amino acid sequence of the mutant human KRAS, the amino acid sequence of the mutant human HRAS, or the amino acid sequence of the mutant human NRAS. Each of the amino acid sequences of the human KRAS, NRAS, and WT HRAS proteins is 188 or 189 amino acid residues long and has a high degree of identity with each other. For example, the amino acid sequence of the human NRAS WT protein is 86.8% identical to the amino acid sequence of the human KRAS WT protein. Amino acid residues 1-86 of the human NRAS WT protein and the human KRAS WT protein are 100% identical. The amino acid sequence of the human HRAS WT protein is 86.3% identical to the amino acid sequence of the human KRAS WT protein. Amino acid residues 1-94 of the human HRAS WT protein and the human KRAS WT protein are 100% identical. Hereafter, references to "RAS" (mutant or non-mutant (WT)) collectively refer to KRAS, HRAS, and NRAS unless otherwise specified.

[0023] In one embodiment of the invention, the amino acid sequence of the mutant human RAS comprises the amino acid sequence of human RAS with a substitution of glycine at position 12 with aspartic acid, wherein position 12 is determined taking into account the corresponding WT RAS protein. The WT RAS protein may be any of the WT KRAS protein (SEQ ID NO: 9 or 10), the WT HRAS protein (SEQ ID NO: 11), or the WT NRAS protein (SEQ ID NO: 12), since, as explained above, amino acid residues 1-86 of the WT human NRAS protein and the WT human KRAS protein are 100% identical, and amino acid residues 1-94 of the WT human HRAS protein and the WT human KRAS protein are 100% identical. Accordingly, the amino acid residue at position 12 of each of the WT KRAS, WT HRAS, and WT NRAS proteins is the same, namely, glycine.

[0024] The amino acid sequence of the mutant human RAS has a substitution of glycine at position 12 with aspartic acid. In this regard, embodiments of the invention provide TCRs with antigen specificity for the amino acid sequence of any human RAS protein, polypeptide, or peptide with the G12D mutation.

[0025] RAS mutations and substitutions are defined herein based on the amino acid sequence of the corresponding WT RAS protein. Thus, RAS mutations and substitutions are described herein based on the amino acid residue located at a particular position in the WT RAS protein (namely, position 12) followed by a position number, followed by the amino acid residue that replaces that residue in the particular mutation or substitution being discussed. An amino acid sequence of RAS (e.g., a RAS peptide) may contain fewer than all of the amino acid residues of the full-length WT RAS protein. Accordingly, position 12 is defined herein based on the full-length WT RAS protein (namely, any of SEQ ID NOs: 9-12), with the understanding that the actual position of the corresponding residue in a particular example of an RAS amino acid sequence may differ. When the positions are as defined by any of SEQ ID NOs: 9-12, the term "G12" refers to the glycine typically found at position 12 of any of SEQ ID NOs: 9-12, and "G12D" indicates that the glycine typically found at position 12 of any of SEQ ID NOs: 9-12 is replaced by aspartic acid. For example, when a particular example of an amino acid sequence of RAS is, for example, WVGAGGVGK (SEQ ID NO: 31) (an illustrative WT KRAS peptide corresponding to contiguous amino acid residues 7-16 of SEQ ID NO: 9), "G12D" refers to the replacement of the underlined glycine in SEQ ID NO: 31 with aspartic acid, even though the actual position of the underlined glycine in SEQ ID NO: 31 is 6. Amino acid sequences of human RAS with the G12D mutation are referred to herein as "G12D RAS".

[0026] Examples of full-length RAS proteins with the G12D mutation are shown below in Table 1.

[0027] In one embodiment of the invention, the TCR has antigen specificity for a RAS peptide with the G12D mutation described above, wherein the RAS G12D peptide is of any length. In one embodiment of the invention, the RAS G12D peptide is of any length suitable for binding to any of the HLA Class I molecules described herein. For example, the TCR may have antigen specificity for a RAS peptide with the G12D mutation, wherein the RAS peptide is from about 9 to about 10 amino acid residues in length. The RAS G12D peptide may comprise any contiguous amino acid residues of a mutant RAS protein that include the G12D mutation. In one embodiment of the invention, the TCR may have antigen specificity for a RAS peptide with the G12D mutation, wherein the mutant RAS peptide is about 9 amino acid residues in length or about 10 amino acid residues in length. Examples of specific peptides, each with a G12D mutation, that can be recognized by the TCR of the invention are 9-mer (SEQ ID NO: 28) and 10-mer (SEQ ID NO: 29). In one embodiment of the invention, the TCR has antigen specificity for the mutant human RAS amino acid sequence of SEQ ID NO: 29. In one embodiment of the invention, the TCR does not have antigen specificity for the wild-type human RAS amino acid sequence WGAGGVGK (SEQ ID NO: 30) or 10-mer (SEQ ID NO: 31).

[0028] In one embodiment of the invention, the TCRs of the invention can recognize G12D RAS presented by an HLA Class I molecule. In this regard, the TCR can elicit an immune response when binding to G12D RAS in the context of an HLA Class I molecule. The TCRs of the invention can recognize G12D RAS that is presented by an HLA Class I molecule and can bind to an HLA Class I molecule in addition to G12D RAS.

[0029] In one embodiment of the invention, the HLA Class I molecule is an HLA-A molecule. The HLA-A molecule is a heterodimer of the α chain and β2 microglobulin. The α chain of HLA-A may be encoded by the HLA-A gene. β2 microglobulin binds non-covalently to the α1, α2, and α3 domains of the α chain to form an HLA-A complex. The HLA-A molecule may be any HLA-A molecule. In one embodiment of the invention, the HLA Class I molecule is an HLA-A11 molecule. The HLA-A11 molecule may be an HLA-A11 molecule. Examples of HLA-A11 molecules may include, but are not limited to, molecules encoded by the HLA-A*11:01, HLA-A*11:02, HLA-A*11:03, or HLA-A*11:04 alleles. Preferably, the HLA Class I molecule is encoded by the HLA-A*11:01 allele.

[0030] The TCRs of the invention may provide any one or more of a variety of advantages, including when expressed by cells used for adoptive cell transfer. G12D RAS is expressed by cancer cells and is not expressed by normal, non-cancerous cells. Without being bound by a particular theory or mechanism, it is believed that the TCRs of the invention advantageously target the destruction of cancer cells while minimizing or eliminating the destruction of normal, non-cancerous cells, thereby reducing toxicity. Furthermore, since the G12D mutation is likely to occur early in tumorigenesis, the G12D RAS mutation may be expressed in substantially all of a patient's cancer cells. The TCRs of the invention may advantageously successfully treat or prevent G12D RAS-positive cancers that are unresponsive to other types of treatment, such as, for example, chemotherapy, surgery, or radiation therapy. Furthermore, the TCRs of the invention can provide high-avidity recognition of G12D RAS, which may provide the ability to recognize unmanipulated tumor cells (e.g., tumor cells that have not been treated with interferon (IFN)-γ, transfected with a vector encoding one or both of G12D RAS and HLA-A*11:01, and pulsed with a G12D RAS peptide or a combination thereof). KRAS mutations are found in approximately 70% of pancreatic cancer, 36% of colorectal cancer, and 20% of lung cancer. The most common mutation is codon 12 (encoding glycine, G). The KRAS G12D mutation is found in approximately 36% and approximately 12% of patients with pancreatic cancer and colorectal cancer, respectively. Furthermore, the HLA-A*11:01 allele is expressed in approximately 14% and approximately 9% of Caucasian and Hispanic ethnic groups, respectively. The HLA-A*11:01 allele is expressed in up to approximately 45% of Asian ethnic groups in the United States. Accordingly, the TCRs of the invention can increase the number of cancer patients eligible for immunotherapy by including those patients expressing the HLA-A*11:01 allele who may not be eligible for immunotherapy using TCRs that recognize RAS presented by other MHC (major histocompatibility complex) molecules. In addition, the TCRs, polypeptides and proteins of the invention contain amino acid sequences of human complementarity determining regions (CDRs) and variable regions, which can reduce the risk of rejection by the human immune system, compared to, for example, TCRs, polypeptides and proteins containing amino acid sequences of mouse CDRs and variable regions.

[0031] The phrase "antigen specificity," as used herein, means that the TCR can specifically bind to and immunologically recognize G12D RAS with high avidity. For example, a TCR is considered to have “antigen specificity” for G12D RAS if about 1 x 10 ⁴ to about 1 x 10 ⁵ T cells expressing the TCR secrete at least about 200 pg/mL or more (e.g., 200 pg/mL or more, 300 pg/mL or more, 400 pg/mL or more, 500 pg/mL or more, 600 pg/mL or more, 700 pg/mL or more, 1,000 pg/mL or more, 5,000 pg/mL or more, 7,000 pg/mL or more, 10,000 pg/mL or more, 20,000 pg/mL or more, or a range defined by any two of the above values) of IFN-γ when co-cultured. with (a) antigen-negative target cells positive for an HLA Class I molecule that have been pulsed with a low concentration of a G12D RAS peptide (e.g., from about 0.05 ng/ml to about 10 ng/ml, 1 ng/ml, 2 ng/ml, 5 ng/ml, 8 ng/ml, 10 ng/ml, or a range defined by any two of the above values), or (b) antigen-negative target cells positive for an HLA Class I molecule that have been pulsed with a nucleotide sequence encoding a G12D RAS such that the target cell expresses a G12D RAS. Cells expressing the TCR of the invention can also secrete IFN-γ when co-cultured with antigen-negative target cells positive for an HLA Class I molecule that have been pulsed with high concentrations of the RAS G12D peptide. The HLA Class I molecule can be any of the HLA Class I molecules described herein (e.g., the HLA-A*11:01 molecule).

[0032] Alternatively, or in addition, a TCR is considered to have "antigen specificity" for G12D RAS if T cells expressing the TCR secrete at least twofold (e.g., fivefold) more IFN-γ when co-cultured with (a) antigen-negative target cells positive for an HLA Class I molecule that have been pulsed with a low concentration of a G12D RAS peptide, or (b) antigen-negative target cells positive for an HLA Class I molecule that have been pulsed with a nucleotide sequence encoding G12D RAS, such that the target cell expresses G12D RAS, compared to the amount of IFN-γ expressed by the negative control. A negative control may be, for example, (i) TCR-expressing T cells co-cultured with (a) antigen-negative target cells positive for an HLA Class I molecule that have been pulsed with the same concentration of an irrelevant peptide (e.g., some other peptide with a sequence different from the G12D RAS peptide), or (b) antigen-negative target cells positive for an HLA Class I molecule that have been pulsed with a nucleotide sequence encoding an irrelevant peptide such that the target cell expresses the irrelevant peptide, or (ii) untransduced T cells (e.g., those derived from PBMCs, which do not express the TCR) co-cultured with (a) target cells (b) antigen-negative, HLA Class I molecule-positive target cells pulsed with the same concentration of G12D RAS peptide, or (c) antigen-negative, HLA Class I molecule-positive target cells pulsed with a nucleotide sequence encoding G12D RAS such that the target cell expresses G12D RAS. The HLA Class I molecule expressed by the negative control target cells could be the same HLA Class I molecule expressed by the target cells co-cultured with the test T cells. The HLA Class I molecule may be any of the HLA Class I molecules described herein (e.g., an HLA-A*11:01 molecule). IFN-γ secretion can be measured by methods known in the art, such as enzyme-linked immunosorbent assay (ELISA).

[0033] Alternatively, or in addition, a TCR is considered to have "antigenic activity" for G12D RAS if at least two-fold (e.g., five-fold) more T cells expressing the TCR secrete IFN-γ when co-cultured with (a) antigen-negative target cells positive for an HLA Class I molecule that have been pulsed with a low concentration of a G12D RAS peptide, or (b) antigen-negative target cells positive for an HLA Class I molecule that have been pulsed with a nucleotide sequence encoding G12D RAS such that the target cell expresses G12D RAS, compared to the number of negative control T cells that secrete IFN-γ. The HLA Class I molecule, peptide concentration, and negative control may be as described herein with respect to other aspects of the invention. The number of IFN-γ-secreting cells may be measured by methods known in the art, such as, for example, ELISPOT.

[0034] Alternatively, or in addition, a TCR is considered to have "antigenic activity" with respect to G12D RAS if T cells expressing the TCR upregulate the expression of one or more markers of T cell activation, as measured, for example, by flow cytometry after stimulation with target cells expressing G12D RAS. Examples of markers of T cell activation include 4-1 BB, OX40, CD107a, CD69, and cytokines that are upregulated upon antigenic stimulation (e.g., tumor necrosis factor (TNF), interleukin (IL)-2, etc.).

[0035] According to an embodiment of the invention, there is provided a TCR comprising two polypeptides (e.g., polypeptide chains), such as a TCR alpha (α) chain, a TCR beta (β) chain, a TCR gamma (γ) chain, a TCR delta (β) chain, or a combination thereof. The TCR polypeptides of the invention may comprise any amino acid sequence, provided that the TCR has antigen specificity for G12D RAS. In some embodiments, the TCR is not naturally occurring.

[0036] In one embodiment of the invention, the TCR comprises two polypeptide chains, each of which comprises a variable region comprising a complementarity determining region (CDR)1, CDR2, and CDR3 of the TCR. In one embodiment of the invention, the TCR comprises a first polypeptide chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO: 1 (CDR1 of the α 4373 TCR chain), a CDR2 comprising the amino acid sequence of SEQ ID NO: 2 (CDR2 of the α 4373 TCR chain), and a CDR3 comprising the amino acid sequence of SEQ ID NO: 3 (CDR3 of the α 4373 TCR chain), and a second polypeptide chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO: 4 (CDR1 of the β 4373 TCR chain), a CDR2 comprising the amino acid sequence of SEQ ID NO: 5 (CDR2 of the β 4373 TCR chain) and a CDR3 comprising the amino acid sequence of SEQ ID NO: 6 (CDR3 of the β 4373 TCR chain).

[0037] In this regard, the TCR of the invention may comprise any one or more amino acid sequences selected from any of SEQ ID NOs: 1-6. In one embodiment of the invention, the TCR comprises the following amino acid sequences: (a) all of SEQ ID NOs: 1-3, (b) all of SEQ ID NOs: 4-6, or (c) all of SEQ ID NOs: 1-6. In a particularly preferred embodiment, the TCR comprises the amino acid sequences of all of SEQ ID NOs: 1-6.

[0038] CDR3 of SEQ ID NO: 3 or 6, namely, the α chain or the β chain or both, may further comprise a cysteine immediately N-terminal to the first amino acid of the CDR, or a phenylalanine immediately C-terminal to the last amino acid, or both.

[0039] In one embodiment of the invention, the TCR comprises an amino acid sequence of a TCR variable region comprising the CDRs set forth above. In this regard, the TCR may, for example, comprise the amino acid sequence of: SEQ ID NO: 7 (variable region of the α4373 TCR chain with the N-terminal signal peptide of the wild type); SEQ ID NO: 51 (variable region of the α4373 TCR chain with the N-terminal signal peptide of the variant); SEQ ID NO: 8 (variable region of the β4373 TCR chain with the N-terminal signal peptide of the variant); SEQ ID NO: 52 (variable region of the β4373 TCR chain with the N-terminal signal peptide of the wild type); SEQ ID NO: 32 (4373 TCR α chain variable region without N-terminal signal peptide, predicted by IMGT); SEQ ID NO: 33 (4373 TCR β chain variable region without N-terminal signal peptide, predicted by IMGT); SEQ ID NO: 59 (4373 TCR α chain variable region without N-terminal signal peptide, predicted by SignalP); SEQ ID NO: 60 (4373 TCR β chain variable region without N-terminal signal peptide, predicted by SignalP); both SEQ ID NOs: 7 and 8; both SEQ ID NOs: 7 and 52; both SEQ ID NOs: 51 and 8; both SEQ ID NOs: 51 and 52; both SEQ ID NOs: 32 and 33, or both SEQ ID NOs: 59 and 60. Preferably, the TCR comprises the following amino acid sequences: (i) both SEQ ID NOs: 7 and 8, (ii) both SEQ ID NOs: 51 and 52, or (iii) both SEQ ID NOs: 32 and 33.

[0040] The TCRs of the invention may further comprise an α chain constant region and a β chain constant region. The constant region may be from any suitable species, such as, for example, a human or a mouse. In one embodiment of the invention, the TCR further comprises murine α and β chain constant regions or human α and β chain constant regions. As used herein, the term "mouse" or "human" when referring to a TCR or any component of a TCR described herein (e.g., a CDR, variable region, constant region, α chain and/or β chain) means a TCR (or component thereof) that is from a mouse or a human, respectively, namely, a TCR (or component thereof) that is derived from or was simultaneously expressed by a murine T cell or a human T cell, respectively.

[0041] According to one embodiment of the invention, there is provided a chimeric TCR comprising a human variable region and a murine constant region, wherein the TCR has antigen specificity for the amino acid sequence of a mutant human RAS with a substitution of glycine at position 12 with aspartic acid, presented by an HLA Class I molecule. The murine constant region may provide one or more advantages. For example, the murine constant region may reduce mispairing of the TCR of the invention with endogenous TCRs of a host cell into which the TCR of the invention is introduced. Alternatively or additionally, the murine constant region may increase the expression level of the TCR of the invention, compared to the same TCR with a human constant region. A chimeric TCR may comprise the amino acid sequence of SEQ ID NO: 19 (mouse WT α chain constant region), SEQ ID NO: 20 (mouse WT β chain constant region), or both SEQ ID NO: 19 and 20. Preferably, the TCR of the invention comprises the amino acid sequences of both SEQ ID NO: 19 and 20. The chimeric TCR may comprise any of the mouse constant regions described herein in combination with any of the CDR regions as described herein in relation to other aspects of the invention. In this regard, the TCR may, for example, comprise the following amino acid sequences: (a) all of SEQ ID NO: 1-3 and 19; (b) all of SEQ ID NO: 4-6 and 20; or (c) all of SEQ ID NO: 1-6 and 19-20. In another embodiment of the invention, the chimeric TCR may comprise any of the murine constant regions described herein in combination with any of the variable regions described herein in relation to other aspects of the invention. In this regard, the TCR may, for example, comprise the following amino acid sequences: (i) both of SEQ ID NOs: 7 and 19; (ii) both of SEQ ID NOs: 51 and 19; (iii) both of SEQ ID NOs: 8 and 20; (iv) both of SEQ ID NOs: 52 and 20; (v) both of SEQ ID NOs: 7-8 and 19-20, or (iv) both of SEQ ID NOs: 51-52 and 19-20.

[0042] In one embodiment of the invention, the TCR comprises an α chain comprising a variable region and a constant region, and a β chain comprising a variable region and a constant region. In this regard, the TCR may, for example, comprise (a) an α chain comprising the amino acid sequence of SEQ ID NO: 21 (the 4373 TCR α chain with a wild-type N-terminal signal peptide), wherein: (i) X at position 193 of SEQ ID NO: 21 is Thr or Cys; (ii) X at position 257 of SEQ ID NO: 21 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 259 of SEQ ID NO: 21 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and (iv) X at position 260 of SEQ ID NO: 21 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) an α chain comprising the amino acid sequence of SEQ ID NO: 53 (4373 TCR α chain with variant N-terminal signal peptide), wherein: (i) X at position 193 of SEQ ID NO: 53 is Thr or Cys; (ii) X at position 257 of SEQ ID NO: 53 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 259 of SEQ ID NO: 53 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and (iv) X at position 260 of SEQ ID NO: 53 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (c) a β chain comprising the amino acid sequence of SEQ ID NO: 22 (4373 TCR β chain with variant N-terminal signal peptide), wherein X at position 191 of SEQ ID NO: 22 is Ser or Cys; (d) a β chain comprising the amino acid sequence of SEQ ID NO: 54 (4373 TCR β chain with wild-type N-terminal signal peptide), wherein X at position 191 of SEQ ID NO: 54 is Ser or Cys; (e) both (a) and (c), (a) and (d), (b) and (c), or (b) and (d); (f) an α chain comprising the amino acid sequence of SEQ ID NO: 34 (the 4373 TCR α chain without the N-terminal signal peptide predicted by IMGT), wherein: (i) the X at position 165 of SEQ ID NO: 34 is Thr or Cys; (ii) the X at position 229 of SEQ ID NO: 34 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) the X at position 231 of SEQ ID NO: 34 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and (iv) the X at position 232 of SEQ ID NO: 34 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (g) a β chain comprising the amino acid sequence of SEQ ID NO: 35 (the 4373 TCR β chain without the N-terminal signal peptide predicted by IMGT), wherein X at position 172 of SEQ ID NO: 35 is Ser or Cys; (h) an α chain comprising the amino acid sequence of SEQ ID NO: 61 (the 4373 TCR α chain without the N-terminal signal peptide predicted by SignalP), wherein: (i) the X at position 172 of SEQ ID NO: 61 is Thr or Cys; (ii) the X at position 236 of SEQ ID NO: 61 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 238 of SEQ ID NO: 61 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and (iv) X at position 239 of SEQ ID NO: 61 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (i) a β chain comprising the amino acid sequence of SEQ ID NO: 62 (the 4373 TCR β chain without the N-terminal signal peptide predicted by SignalP), wherein X at position 170 of SEQ ID NO: 62 is Ser or Sue; or (j), both (f) and (g), or both (h) and (i).

[0043] In another embodiment of the invention, the TCR comprises the amino acid sequence(s): SEQ ID NO: 23 (α4373 TCR chain with WT mouse constant region and WT N-terminal signal peptide), SEQ ID NO: 55 (wild-type mouse constant region of the α4373 TCR chain and variant N-terminal signal peptide), SEQ ID NO: 24 (β4373 TCR chain with wild-type mouse constant region and variant N-terminal signal peptide), SEQ ID NO: 56 (β4373 TCR chain with WT mouse constant region and wild-type N-terminal signal peptide), SEQ ID NO: 36 (α4373 TCR chain with WT mouse constant region and without N-terminal signal peptide, predicted by IMGT), SEQ ID NO: 37 (β4373 TCR chain with mouse WT constant region and without N-terminal signal peptide, predicted by IMGT), SEQ ID NO: 63 (α4373 TCR chain with mouse WT constant region and without N-terminal signal peptide, predicted by SignalP), SEQ ID NO: 64 (β4373 TCR chain with mouse WT constant region and without N-terminal signal peptide, predicted by SignalP), both of SEQ ID NOs: 23 and 24, both of SEQ ID NOs: 55 and 24, both of SEQ ID NOs: 23 and 56, both of SEQ ID NOs: 55 and 56, both of SEQ ID NOs: 36 and 37, or both of SEQ ID NOs: 63 and 64.

[0044] In one embodiment of the invention, the TCR comprises a constant region with a substitution. In this regard, the TCR, for example, may comprise the amino acid sequence of any of the TCRs described herein with one, two, three or four amino acid substitutions in the constant region of one or both of the α and β chains. Preferably, the TCR comprises a murine constant region with one, two, three or four amino acid substitutions in the murine constant region of one or both of the α and β chains. In a particularly preferred embodiment, the TCR comprises a murine constant region with one, two, three or four amino acid substitutions in the murine constant region of the α chain and one amino acid substitution in the murine constant region of the β chain. In some embodiments, TCRs containing the constant region with the substitution advantageously provide one or more of increased recognition of G12D RAS ⁺ targets, increased host cell expression levels, reduced mismatch with endogenous TCRs, and increased anti-tumor activity, compared to a parental TCR containing the constant region without the substitution (wild type). In general, the amino acid sequences with substitution of the mouse constant regions of the ai TCR chains, SEQ ID NO: 17 and 18, respectively, correspond to all or parts of the mouse amino acid sequences of the constant regions without substitutions of SEQ ID NO: 19 and 20, respectively, wherein SEQ ID NO: 17 has one, two, three or four amino acid substitutions compared to SEQ ID NO: 19, and SEQ ID NO: 18 has one amino acid substitution compared to SEQ ID NO: 20. In connection with this, according to an embodiment of the invention there is provided a TCR comprising the amino acid sequences of (a) SEQ ID NO: 17 (the constant region of the α chain), wherein (i) X at position 48 is Thr or Cys; (ii) X at position 112 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 114 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and (iv) X at position 115 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) SEQ ID NO: 18 (β chain constant region), wherein X at position 57 is Ser or Cys; or (c) both of SEQ ID NOs: 17 and 18. In one embodiment of the invention, a TCR comprising SEQ ID NO: 17 does not comprise SEQ ID NO: 19 (mouse constant region without α chain substitution). In one embodiment of the invention, a TCR comprising SEQ ID NO: 18 does not comprise SEQ ID NO: 20 (mouse constant region without β chain substitution).

[0045] The first amino acid of any of the mouse alpha constant regions described herein may differ from N, as provided in SEQ ID NOs: 17 and 19. For example, in any TCR construct, polypeptide, protein, etc., as described herein, this first amino acid may be encoded by a split codon (having nucleotides from both the variable region and the constant region), such that any of the mouse alpha constant regions may have a different amino acid at that position. Similarly, the first amino acid of any of the mouse beta constant regions described herein may differ from E, as provided in SEQ ID NOs: 18 and 20, for example, this first amino acid may be encoded by a split codon.

[0046] In one embodiment of the invention, the substituted constant region comprises cysteine substitutions in the constant region of one or both of the α and β chains to provide a cysteine-substituted TCR. Opposite cysteines in the α and β chains provide a disulfide bond that links the constant regions of the substituted α and β chains of the TCR to each other and which is not present in a TCR comprising murine constant regions without the substitution. In this regard, the TCR, for example, can be a cysteine-substituted TCR in which one or both of the native Thr at position 48 (Thr48) of SEQ ID NO: 19 and the native Ser at position 57 (Ser57) of SEQ ID NO: 20 can be replaced with Cys. Preferably, both of the native Thr48 of SEQ ID NO: 19 and the native Ser57 of SEQ ID NO: 20 are replaced with Cys. Exemplary sequences of constant regions of cysteine-substituted TCRs are set forth in Table 2. In one embodiment of the invention, a cysteine-substituted TCR comprises (i) SEQ ID NO: 17, (ii) SEQ ID NO: 18, or (iii) both of SEQ ID NOs: 17 and 18, where both of SEQ ID NOs: 17 and 18 are as defined in Table 2. A cysteine-substituted TCR of the invention may include a substituted constant region in addition to any of the CDRs or variable regions described herein.

[0047] In one embodiment of the invention, the chimeric TCR with a cysteine substitution comprises a full-length α chain and a full-length β chain. Examples of α chain and β chain sequences of a chimeric TCR with a cysteine substitution are set out in Table 2. In one embodiment of the invention, the TCR comprises (i) SEQ ID NO: 21, (ii) SEQ ID NO: 53, (iii) SEQ ID NO: 22, (iv) SEQ ID NO: 54, (v) SEQ ID NO: 34, (vi) SEQ ID NO: 35, (vii) SEQ ID NO: 61, (viii) SEQ ID NO: 62, (ix) both of SEQ ID NOs: 21 and 22, (x) both of SEQ ID NOs: 53 and 22, (xi) both of SEQ ID NOs: 21 and 54, (xii) both of SEQ ID NOs: 53 and 54, (xiii) both of SEQ ID NOs: 34 and 35, or (xiv) both of SEQ ID NO: 61 and 62, wherein all of SEQ ID NO: 21-22, 34-35, 53, 54, 61, and 62 are as defined in Table 2.

[0048] In one embodiment of the invention, the substitution amino acid sequence comprises substitutions of one, two, or three amino acids in the transmembrane (TM) domain of the α chain constant region with a hydrophobic amino acid to provide a TCR with a hydrophobic amino acid substitution (also referred to herein as an "LVL-modified TCR"). The hydrophobic amino acid substitution(s) in the TM domain of the TCR may increase the hydrophobicity of the TM domain of the TCR, compared to a TCR that does not have the hydrophobic amino acid substitution(s) in the TM domain. In this regard, the TCR is an LVL-modified TCR in which one, two, or three of the native Ser112, Met114, and Gly115 of SEQ ID NO: 19 may, independently, be replaced with Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; preferably Leu, Ile, or Val; and native Ser57 of SEQ ID NO: 20 may be replaced with Cys. Preferably, all three of native Ser112, Met114 and Gly115 of SEQ ID NO: 19 may, independently, be replaced with Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; preferably Leu, Ile or Val. In one embodiment of the invention, the LVL-modified TCR comprises (i) SEQ ID NO: 17, (ii) SEQ ID NO: 18 or (iii) both SEQ ID NOs: 17 and 18, wherein both SEQ ID NOs: 17 and 18 are as defined in Table 3. The LVL-modified TCRs of the invention may include a constant region with a substitution in addition to any of the CDRs or variable regions described herein.

[0049] In one embodiment of the invention, the LVL-modified TCR comprises a full-length α chain and a full-length β chain. Examples of sequences of the α chain and the β chain of the LVL-modified TCR are set out in Table 3. In one embodiment of the invention, the LVL-modified TCR comprises (i) SEQ ID NO: 21, (ii) SEQ ID NO: 53, (iii) SEQ ID NO: 22, (iv) SEQ ID NO: 54, (v) SEQ ID NO: 34, (vi) SEQ ID NO: 35, (vii) SEQ ID NO: 61, (viii) SEQ ID NO: 62, (ix) both of SEQ ID NOs: 21 and 22, (x) both of SEQ ID NOs: 53 and 22, (xi) both of SEQ ID NOs: 21 and 54, (xii) both of SEQ ID NOs: 53 and 54, (xiii) both of SEQ ID NOs: 34 and 35, or (xiv) both of SEQ ID NO: 61 and 62, wherein all of SEQ ID NO: 21-22, 34-35, 53, 54, 61, and 62 are as defined in Table 3.

[0050] In one embodiment of the invention, the substitution amino acid sequence comprises cysteine substitutions in the constant region of one or both of the α and β chains in combination with the substitution(s) of one, two, or three amino acids in the transmembrane (TM) domain of the α chain constant region with a hydrophobic amino acid (also referred to herein as an "LVL-modified cysteine substitution TCR"). In this regard, the TCR is an LVL-modified, chimeric cysteine substitution TCR in which native Thr48 of SEQ ID NO: 19 is replaced with Cys; one, two, or three of native Ser112, Met114, and Gly115 of SEQ ID NO: 19, independently, are replaced by Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; preferably, Leu, Ile, or Val; and native Ser57 of SEQ ID NO: 20 is replaced with Cys. Preferably, all three of native Ser112, Met114 and Gly115 of SEQ ID NO: 19 may, independently, be replaced with Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; preferably, Leu, Ile or Val. In one embodiment of the invention, the cysteine substitution LVL-modified TCR comprises (i) SEQ ID NO: 17, (ii) SEQ ID NO: 18 or (iii) both of SEQ ID NOs: 17 and 18, wherein both SEQ ID NOs: 17 and 18 are as defined in Table 4. The cysteine substitution LVL-modified TCRs of the invention may include a constant region with a substitution in addition to any of the CDRs or variable regions described herein.

[0051] In one embodiment, the LVL-modified cysteine substitution TCR comprises a full-length α chain and a full-length β chain. In one embodiment of the invention, the LVL-modified TCR with a cysteine substitution comprises (i) SEQ ID NO: 21, (ii) SEQ ID NO: 53, (iii) SEQ ID NO: 22, (iv) SEQ ID NO: 54, (v) SEQ ID NO: 34, (vi) SEQ ID NO: 35, (vii) SEQ ID NO: 61, (viii) SEQ ID NO: 62, (ix) both of SEQ ID NOs: 21 and 22, (x) both of SEQ ID NOs: 53 and 22, (xi) both of SEQ ID NOs: 21 and 54, (xii) both of SEQ ID NOs: 53 and 54, (xiii) both of SEQ ID NOs: 34 and 35, or (xiv) both of SEQ ID NOs: 61 and 62, wherein all of SEQ ID NOs: 21-22, 34-35, 53, 54, 61 and 62 are as defined in Table 4.

[0052] In one embodiment of the invention, the LVL-modified TCR with a cysteine substitution comprises: (a) SEQ ID NO: 38 (LVL-modified TCR α chain constant region with a cysteine substitution); (b) SEQ ID NO: 39 (LVL-modified TCR β chain constant region with a cysteine substitution); (c) SEQ ID NO: 40 (LVL-modified 4373 TCR α chain with a wild-type N-terminal signal sequence); (d) SEQ ID NO: 41 (LVL-modified 4373 TCR β chain with a variant N-terminal signal sequence); (e) SEQ ID NO: 42 (α chain of LVL-modified 4373 TCR with cysteine substitution without N-terminal signal sequence predicted by IMGT); (f) SEQ ID NO: 43 (β chain of LVL-modified 4373 TCR with cysteine substitution without N-terminal signal sequence predicted by IMGT); (g) SEQ ID NO: 65 (α chain of LVL-modified 4373 TCR with cysteine substitution without N-terminal signal sequence predicted by SignalP); (h) SEQ ID NO: 66 (β chain of LVL-modified 4373 TCR with cysteine substitution without N-terminal signal sequence predicted by SignalP); (i) SEQ ID NO: 57 (α chain of LVL-modified 4373 TCR with cysteine substitution variant N-terminal signal sequence); (j) SEQ ID NO: 58 (β chain of LVL-modified 4373 TCR with cysteine substitution wild-type N-terminal signal sequence); (k) both (a) and (b); (i) both (c) and (d); (m) both (e) and (f); (n) both (g) and (h); or (o) both (i) and (j).

[0053] Also provided according to the invention is a polypeptide comprising a functional portion of any of the TCRs described herein. The term "polypeptide," as used herein, includes oligopeptides and refers to a single chain of amino acids linked by one or more peptide bonds.

[0054] With respect to the polypeptides of the invention, a functional portion may be any portion that contains contiguous amino acids of the TCR of which it is a part, so long as the functional portion specifically binds to G12D RAS. The term "functional portion," when used in relation to a TCR, refers to any portion or fragment of a TCR of the invention whose portion or fragment retains the biological activity of the TCR of which it is a part (the parent TCR). Functional portions include, for example, those portions of the TCR that retain the ability to specifically bind to G12D RAS (e.g., in the context of the HLA-A*11:01 molecule), or to detect, treat, or prevent cancer to a similar, equal, or greater extent than the parent TCR. With respect to the original TCR, the functional portion may comprise, for example, about 10%, about 25%, about 30%, about 50%, about 68%, about 80%, about 90%, about 95% or more of the original TCR.

[0055] The functional portion may comprise additional amino acids at the N- or C-terminus of the portion, or at both ends, additional amino acids that are not found in the amino acid sequence of the parent TCR. Desirably, the additional amino acids do not interfere with the biological function of the functional portion, such as specific binding to G12D RAS; and/or having the ability to detect cancer, treat or prevent cancer, etc. More desirably, the additional amino acids enhance the biological activity, compared to the biological activity of the parent TCR.

[0056] The polypeptide may comprise a functional portion of either or both of the α and β chains of the TCR of the invention, such as a functional portion comprising one or more of CDR1, CDR2, and CDR3 of the variable region(s) of the α chain and/or the β chain of the TCR of the invention. In one embodiment of the invention, the polypeptide may comprise the amino acid sequence of SEQ ID NO: 1 (CDR1 of the α chain), SEQ ID NO: 2 (CDR2 of the α chain), SEQ ID NO: 3 (CDR3 of the α chain), SEQ ID NO: 4 (CDR1 of the β chain), SEQ ID NO: 5 (CDR2 of the β chain), SEQ ID NO: 6 (CDR3 of the β chain), or a combination thereof.

[0057] In this regard, the polypeptide of the invention may comprise any one or more amino acid sequences selected from any one of SEQ ID NOs: 1-6. In one embodiment of the invention, the TCR comprises the following amino acid sequences: (a) all of SEQ ID NOs: 1-3, (b) all of SEQ ID NOs: 4-6, or (c) all of SEQ ID NOs: 1-6. In a preferred embodiment, the polypeptide comprises the amino acid sequences of all of SEQ ID NOs: 1-6. CDR3 of SEQ ID NO: 3 or 6, namely, the α chain or the β chain or both, may further comprise a cysteine immediately N-terminal to the first amino acid of the CDR, or a phenylalanine immediately C-terminal to the final amino acid, or both.

[0058] In one embodiment of the invention, a polypeptide of the invention may comprise, for example, a TCR variable region of the invention comprising a combination of the CDR regions set forth above. In this regard, the polypeptide may comprise the amino acid sequence of (i) SEQ ID NO: 7 (α chain variable region with wild-type N-terminal signal sequence), (ii) SEQ ID NO: 51 (4373 TCR α chain variable region with variant N-terminal signal sequence); (iii) SEQ ID NO: 8 (β chain variable region with variant N-terminal signal sequence), (iv) SEQ ID NO: 52 (β chain variable region with wild-type N-terminal signal sequence), (v) both of SEQ ID NOs: 7 and 8; (vi) both of SEQ ID NOs: 51 and 8, (vii) both of SEQ ID NOs: 7 and 52, or (viii) both of SEQ ID NOs: 51 and 52, (ix) SEQ ID NO: 32 (the α chain variable region without the N-terminal signal sequence, predicted by IMGT), (x) SEQ ID NO: 33 (the β chain variable region without the N-terminal signal sequence, predicted by IMGT), (xi) SEQ ID NO: 59 (the 4373 TCR α chain variable region without the N-terminal signal peptide, predicted by SignalP); (xii) SEQ ID NO: 60 (the 4373 TCR β chain variable region without the N-terminal signal peptide, predicted by SignalP); (xiii) both of SEQ ID NO: 32 and 33 or both of SEQ ID NO: 59 and 60. Preferably, the polypeptide comprises the following amino acid sequences: (i) both of SEQ ID NO: 7 and 8, (ii) both of SEQ ID NO: 51 and 52, (iii) both of SEQ ID NO: 32 and 33, or (iv) both of SEQ ID NO: 59 and 60.

[0059] In one embodiment of the invention, the polypeptide of the invention may further comprise a TCR constant region of the invention as set forth above. In this regard, the polypeptide may further comprise the amino acid sequence of SEQ ID NO: 19 (WT mouse α chain constant region), SEQ ID NO: 20 (WT mouse β chain constant region), SEQ ID NO: 17 (mouse α chain substitution constant region), SEQ ID NO: 18 (mouse β chain substitution constant region), SEQ ID NO: 38 (LVL-modified TCR α chain constant region with cysteine substitution); SEQ ID NO: 39 (LVL-modified TCR β chain constant region with cysteine substitution); both SEQ ID NOs: 19 and 20, both SEQ ID NOs: 17 and 18, or both SEQ ID NOs: 38 and 39. Preferably, the polypeptide further comprises the amino acid sequences of both SEQ ID NOs: 17 and 18, both SEQ ID NOs: 19 and 20, or both SEQ ID NOs: 38 and 39 in combination with any of the CDR regions or variable regions described herein, in relation to other aspects of the invention. In one embodiment of the invention, one or both of SEQ ID NOs: 17 and 18 of the polypeptide are as defined in any of Tables 2-4. The α chain constant regions provided herein are shown with an N-terminal asparagine. In some embodiments, the N-terminal amino acid of the α chain constant regions described herein is aspartic acid.

[0060] In one embodiment of the invention, the polypeptide of the invention may comprise the full length chain of an α or β TCR described herein. In this regard, the polypeptide of the invention may comprise the amino acid sequence of SEQ ID NO: 21, SEQ ID NO: 53, SEQ ID NO: 22, SEQ ID NO: 54, SEQ ID NO: 23, SEQ ID NO: 55, SEQ ID NO: 24, SEQ ID NO: 56, SEQ ID NO: 34, SEQ ID NO: 61, SEQ ID NO: 35, SEQ ID NO: 62, SEQ ID NO: 36, SEQ ID NO: 63, SEQ ID NO: 37, SEQ ID NO: 64, SEQ ID NO: 40, SEQ ID NO: 57, SEQ ID NO: 41, SEQ ID NO: 58, SEQ ID NO: 42, SEQ ID NO: 65, SEQ ID NO: 43, SEQ ID NO: 66, both of SEQ ID NO: 21-22, both of SEQ ID NOs: 21 and 54, both of SEQ ID NOs: 53 and 22, both of SEQ ID NOs: 53 and 54, both of SEQ ID NOs: 23-24, both of SEQ ID NOs: 55 and 24, both of SEQ ID NOs: 23 and 54, both of SEQ ID NOs: 55 and 54, both of SEQ ID NO: 34-35, both from SEQ ID NO: 36-37, both from SEQ ID NO: 40-41, both from SEQ ID NO: 57 and 41, both from SEQ ID NO: 40-58, both from SEQ ID NO: 57-58, both from SEQ ID NO: 42-43, both from SEQ ID NO: 61 and 62, both from SEQ ID NO: 63 and 64 or both of SEQ ID NO: 65 and 66. Alternatively, the polypeptide of the invention may comprise both TCR chains described herein.

[0061] For example, a polypeptide of the invention may comprise (a) the amino acid sequence of SEQ ID NO: 21, wherein: (i) the X at position 193 of SEQ ID NO: 21 is Thr or Cys; (ii) the X at position 257 of SEQ ID NO: 21 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) the X at position 259 of SEQ ID NO: 21 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and (iv) the X at position 260 of SEQ ID NO: 21 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) the amino acid sequence of SEQ ID NO: 53, wherein: (i) the X at position 193 of SEQ ID NO: 53 is Thr or Cys; (ii) X at position 257 of SEQ ID NO: 53 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 259 of SEQ ID NO: 53 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and (iv) X at position 260 of SEQ ID NO: 53 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (c) the amino acid sequence of SEQ ID NO: 22, wherein X at position 191 of SEQ ID NO: 22 is Ser or Cys; (d) the amino acid sequence of SEQ ID NO: 54, wherein X at position 191 of SEQ ID NO: 54 is Ser or Cys; (e) both (a) and (c), (a) and (d), (b) and (c), or (b) and (d); (f) the amino acid sequence of SEQ ID NO: 34, wherein: (i) X at position 165 of SEQ ID NO: 34 is Thr or Cys; (ii) X at position 229 of SEQ ID NO: 34 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 231 of SEQ ID NO: 34 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and (iv) X at position 232 of SEQ ID NO: 34 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (g) the amino acid sequence of SEQ ID NO: 35, wherein X at position 172 of SEQ ID NO: 35 is Ser or Cys; (h) the amino acid sequence of SEQ ID NO: 61, wherein: (i) X at position 172 of SEQ ID NO: 61 is Thr or Cys; (ii) X at position 236 of SEQ ID NO: 61 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 238 of SEQ ID NO: 61 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and (iv) X at position 239 of SEQ ID NO: 61 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (i) the amino acid sequence of SEQ ID NO: 62, wherein X at position 170 of SEQ ID NO: 62 is Ser or Cys; (j) both (f) and (g) or both (h) and (i); (k) SEQ ID NO: 40; (I) SEQ ID NO: 57; (m) SEQ ID NO: 41; (n) SEQ ID NO: 58; (o) SEQ ID NO: 42; (p) SEQ ID NO: 43; (q) SEQ ID NO: 65; (r) SEQ ID NO: 66; (s) both (k) and (m); (t) both (I) and (m); (u) both (k) and (n); (v) both (I) and (n); (w) both (o) and (p); or (x) both (q) and (r). In one embodiment of the invention, any one or more of SEQ ID NOs: 21-22, 34-35, 53, 54, 61 and 62 polypeptides are as defined in any of Tables 2-4.

[0062] The invention further provides a protein comprising at least one of the polypeptides described herein. By "protein" is meant a molecule comprising one or more polypeptide chains.

[0063] In one embodiment, the protein of the invention may comprise a first polypeptide chain comprising the amino acid sequences of SEQ ID NOs: 1-3 and a second polypeptide chain comprising the amino acid sequence of SEQ ID NOs: 4-6. CDR3 of SEQ ID NO: 3 or 6, namely, the α chain or the β chain or both, may further comprise a cysteine immediately N-terminal to the first amino acid of the CDR, or a phenylalanine immediately C-terminal to the final amino acid, or both.

[0064] In another embodiment of the invention, (i) the first polypeptide chain of the protein may comprise the amino acid sequence of SEQ ID NO: 7 and the second polypeptide chain may comprise the amino acid sequence of SEQ ID NO: 8; (ii) the first polypeptide chain of the protein may comprise the amino acid sequence of SEQ ID NO: 51 and the second polypeptide chain may comprise the amino acid sequence of SEQ ID NO: 8; (iii) the first polypeptide chain of the protein may comprise the amino acid sequence of SEQ ID NO: 7 and the second polypeptide chain may comprise the amino acid sequence of SEQ ID NO: 52; (iv) the first polypeptide chain of the protein may comprise the amino acid sequence of SEQ ID NO: 51 and the second polypeptide chain may comprise the amino acid sequence of SEQ ID NO: 52; (v) the first polypeptide chain of the protein may comprise the amino acid sequence of SEQ ID NO: 32 and the second polypeptide chain may comprise the amino acid sequence of SEQ ID NO: 33 or (vi) the first polypeptide chain of the protein may comprise the amino acid sequence of SEQ ID NO: 59 and the second polypeptide chain may comprise the amino acid sequence of SEQ ID NO: 60.

[0065] The protein of the invention may further comprise any of the constant regions described herein in relation to other aspects of the invention. Accordingly, in one embodiment of the invention, (i) the first polypeptide chain may further comprise the amino acid sequence of SEQ ID NO: 17 and the second polypeptide chain may further comprise the amino acid sequence of SEQ ID NO: 18; (ii) the first polypeptide chain may further comprise the amino acid sequence of SEQ ID NO: 19 and the second polypeptide chain may further comprise the amino acid sequence of SEQ ID NO: 20; or (ii) the first polypeptide chain may further comprise the amino acid sequence of SEQ ID NO: 38 and the second polypeptide chain may further comprise the amino acid sequence of SEQ ID NO: 39. In one embodiment of the invention, one or both of SEQ ID NOs: 17 and 18 of the protein are as defined in any of Tables 2-4.

[0066] Alternatively or additionally, a protein of the invention may comprise the following: (a) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 21, wherein: (i) X at position 193 of SEQ ID NO: 21 is Thr or Cys; (ii) X at position 257 of SEQ ID NO: 21 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 259 of SEQ ID NO: 21 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and (iv) X at position 260 of SEQ ID NO: 21 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (b) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 53, wherein: (i) X at position 193 of SEQ ID NO: 53 is Thr or Cys; (ii) X at position 257 of SEQ ID NO: 53 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 259 of SEQ ID NO: 53 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 260 of SEQ ID NO: 53 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (c) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 22, wherein X at position 191 of SEQ ID NO: 22 is Ser or Cys; (d) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 54, wherein X at position 191 of SEQ ID NO: 54 is Ser or Cys; (e) both of (a) and (c), (a) and (d), (b) and (c), or (b) and (d); (f) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 34, wherein: (i) X at position 165 of SEQ ID NO: 34 is Thr or Cys; (ii) X at position 229 of SEQ ID NO: 34 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 231 of SEQ ID NO: 34 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and (iv) X at position 232 of SEQ ID NO: 34 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (g) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 35, wherein X at position 172 of SEQ ID NO: 35 is Ser or Cys; (h) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, wherein: (i) X at position 172 of SEQ ID NO: 61 is Thr or Cys; (ii) X at position 236 of SEQ ID NO: 61 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; (iii) X at position 238 of SEQ ID NO: 61 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and (iv) X at position 239 of SEQ ID NO: 61 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (i) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 62 (TCR β4373 chain without the N-terminal signal peptide, predicted by SignalP), wherein X at position 170 of SEQ ID NO: 62 is Ser or Cys or (j) both (f) or both (h) and (i) and; (k) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 40; (k) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 57; (m) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 41; (n) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 58; (o) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 42; (p) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 43; (p) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 65; (q) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 66; (s) both (k) and (m); (t) both (l) and (m); (u) both (k) and (n); (v) both (m) and (n); (w) both (o) and (p) or both (q) and (r). In one embodiment of the invention, one or more of SEQ ID NOs: 21-22, 34-35, 53, 54, 61 and 62 are as defined in any one of Tables 2-4.

[0067] The protein of the invention may be a TCR. Alternatively, if, for example, the protein comprises one polypeptide chain comprising the amino acid sequences of both SEQ ID NOs: 23 and 24, both SEQ ID NOs: 55 and 24, both SEQ ID NOs: 23 and 56, both SEQ ID NOs: 55 and 56, both SEQ ID NOs: 21 and 22, both SEQ ID NOs: 53 and 22, both SEQ ID NOs: 21 and 54, both SEQ ID NOs: 53 and 54, or if the first and/or second polypeptide chain(s) of the protein further comprise other amino acid sequences, such as an amino acid sequence encoding an immunoglobulin or a portion thereof, then the protein of the invention may be a fusion protein. In this regard, the invention also provides a fusion protein comprising at least one of the polypeptides of the invention described herein, together with at least one other polypeptide. The other polypeptide may exist as a separate polypeptide of the fusion protein or may exist as a polypeptide that is expressed in frame (in tandem) with one of the polypeptides of the invention described herein. The other polypeptide may encode any peptide or protein molecule or part thereof, including, but not limited to, an immunoglobulin, CD3, CD4, CD8, an MHC molecule, a CD1 molecule, such as CD1a, CD1b, CD1c, CD1d, etc.

[0068] A fusion protein may comprise one or more copies of a polypeptide of the invention and/or one or more copies of another polypeptide. For example, a fusion protein may comprise 1, 2, 3, 4, 5 or more copies of a polypeptide of the invention and/or another polypeptide. Suitable methods for producing fusion proteins are known in the art and include, for example, genetic engineering methods.

[0069] In some embodiments of the invention, the TCRs, polypeptides, and proteins of the invention can be expressed as a single protein comprising a linker peptide linking the α chain and the β chain. In this regard, the TCRs, polypeptides, and proteins of the invention can further comprise a linker peptide. The linker peptide can advantageously facilitate expression of the recombinant TCR, polypeptide, and/or protein in a host cell. The linker polypeptide can comprise any suitable amino acid sequence. The linker peptide can be a cleavable linker peptide. For example, the linker peptide can be a furin-SGSG-P2A linker comprising the amino acid sequence RAKRSGSGATNFSLLKQAGDVEENPGP (SEQ ID NO: 25). When a construct comprising a linker peptide is expressed by a host cell, the linker peptide may be cleaved, resulting in separated α and β chains. In one embodiment of the invention, a TCR, polypeptide, or protein may comprise an amino acid sequence comprising a full-length α chain, a full-length β chain, and a linker peptide located between the α and β chains, such as α chain-linker-β chain or β chain-linker-α chain.

[0070] In one embodiment of the invention, a TCR, polypeptide, or protein may comprise an amino acid sequence as set forth in SEQ ID NO: 47, comprising, from N-terminus to C-terminus, a β chain, a linker (SEQ ID NO: 25), and an α chain. The variant comprises a β chain variable region (with a variant signal peptide) as set forth in SEQ ID NO: 8 and a modified β constant domain as set forth in SEQ ID NO: 39. The full-length β chain of the variant is set forth in SEQ ID NO: 41. The variant also comprises an α chain variable region (with a wild-type signal peptide) as set forth in SEQ ID NO: 7 and a modified α constant domain as set forth in SEQ ID NO: 38. The full-length α chain of the variant is set forth in SEQ ID NO: 40.

[0071] In another embodiment of the invention, the TCR, polypeptide or protein may comprise an amino acid sequence as set forth in SEQ ID NO: 67, comprising, from N-terminus to C-terminus, an α chain, a linker (SEQ ID NO: 25) and a β chain. The variant comprises an α chain variable region (with a variant signal peptide) as set forth in SEQ ID NO: 51 and a modified α constant domain as set forth in SEQ ID NO: 38. The full-length α chain of the variant is set forth in SEQ ID NO: 57. The variant also comprises a β chain variable region (with a wild-type signal peptide) as set forth in SEQ ID NO: 52 and a modified β constant domain as set forth in SEQ ID NO: 39. The full-length β chain of the variant is set forth in SEQ ID NO: 58.

[0072] In some embodiments, the TCR, polypeptide, or protein disclosed herein comprises an α chain and/or a β chain as disclosed herein, comprising a signal peptide. In some embodiments, the signal peptide sequence of any of the α chains and/or β chains disclosed herein comprises an alanine or histidine residue used in place of the wild-type residue at position 2.

[0073] In some embodiments, the TCR, polypeptide, or protein disclosed herein comprises a mature version of the α chain and/or β chain as disclosed herein that does not have a signal peptide. The signal peptide sequence or the mature form of the α chain and/or β chain can be implemented according to any method known in the art, including IMGT and SignalP.

[0074] The protein of the invention may be a recombinant antibody or an antigen-binding portion thereof that comprises at least one of the polypeptides of the invention disclosed herein. As used herein, the term "recombinant antibody" refers to a recombinant (e.g., genetically engineered) protein that comprises at least one of the polypeptides of the invention and a polypeptide chain of an α-antibody or an antigen-binding portion thereof. The polypeptide of the antibody or an antigen-binding portion thereof may be a heavy chain, a light chain, a variable or constant region of a heavy or light chain, a single-chain variable fragment (scFv), or an Fc, Fab, or F(ab) ₂ ' fragment of an antibody, etc. The polypeptide chain of the α-antibody or an antigen-binding portion thereof may exist as a separate polypeptide of the recombinant antibody. Alternatively, the polypeptide chain of the antibody or antigen-binding portion thereof may exist as a polypeptide that is expressed in frame (in tandem) with the polypeptide of the invention. The polypeptide of the antibody or antigen-binding portion thereof may be a polypeptide of any antibody or any antibody fragment, including any of the antibodies and antibody fragments described herein.

[0075] Included within the scope of the present invention are functional variants of the TCRs, polypeptides, or proteins of the invention described herein. The term "functional variant," as used herein, refers to a TCR, polypeptide, or protein that has substantial or significant sequence identity or similarity to a parent TCR, polypeptide, or protein, the functional variant of which retains the biological activity of the TCR, polypeptide, or protein of which it is a variant. Functional variants include, for example, those variants of the TCR, polypeptide, or protein described herein (the parent TCR, polypeptide, or protein) that retain the ability to specifically bind to the G12D RAS for which the parent TCR has antigen specificity or to which the parent polypeptide or protein specifically binds, to a similar degree, to the same degree, or to a greater degree than the parent TCR, polypeptide, or protein. With respect to the parent TCR, polypeptide, or protein, the functional variant may, for example, be at least about 30%, about 50%, about 75%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or more identical in amino acid sequence to the parent TCR, polypeptide, or protein, respectively.

[0076] A functional variant may, for example, comprise an amino acid sequence of a parent TCR, polypeptide, or protein with at least one conservative amino acid substitution. Conservative amino acid substitutions are known in the art and include amino acid substitutions in which one amino acid having certain physical and/or chemical properties is replaced by another amino acid that has the same chemical or physical properties. For example, a conservative amino acid substitution may be an acidic amino acid replacing another acidic amino acid (e.g., Asp or Glu), an amino acid with a non-polar side chain replacing another amino acid with a non-polar side chain (e.g., Ala, Gly, Val, Ile, Leu, Met, Phe, Pro, Trp, Val, etc.), a basic amino acid replacing another basic amino acid (Lys, Arg, etc.), an amino acid with a polar side chain replacing another amino acid with a polar side chain (Asn, Cys, Gin, Ser, Thr, Tyr, etc.), etc.

[0077] Alternatively or additionally, functional variants may comprise an amino acid sequence of the parent TCR, polypeptide, or protein with at least one non-conservative amino acid substitution. In this case, it is preferred that the non-conservative amino acid substitution does not affect or inhibit the biological activity of the functional variant. Preferably, the non-conservative amino acid substitution enhances the biological activity of the functional variant, such that the biological activity of the functional variant is increased, compared to the parent TCR, polypeptide, or protein.

[0078] Each TCR signal peptide, polypeptide, protein, functional variant, and functional portion described herein, if present, may be any suitable TCR signal peptide, so long as the TCR, polypeptide, protein, or functional variant is expressed and has antigen specificity for the amino acid sequence of a mutant human RAS with a substitution of glycine at position 12 with aspartic acid, presented by an HLA Class I molecule.

[0079] The TCR, polypeptide, or protein may consist essentially of a given amino acid sequence and the sequences described herein, such that other components of the TCR, polypeptide, or protein, such as other amino acids, do not substantially alter the biological activity of the TCR, polypeptide, or protein. In this regard, the TCR, polypeptide or protein of the invention may, for example, consist essentially of the amino acid sequence of SEQ ID NO: 21, SEQ ID NO: 53, SEQ ID NO: 22, SEQ ID NO: 54, SEQ ID NO: 23, SEQ ID NO: 55, SEQ ID NO: 24, SEQ ID NO: 56, SEQ ID NO: 34, SEQ ID NO: 61, SEQ ID NO: 35, SEQ ID NO: 62, SEQ ID NO: 36, SEQ ID NO: 63, SEQ ID NO: 37, SEQ ID NO: 64, SEQ ID NO: 40, SEQ ID NO: 57, SEQ ID NO: 41, SEQ ID NO: 58, SEQ ID NO: 42, SEQ ID NO: 65, SEQ ID NO: 43, SEQ ID NO: 66, both SEQ ID NOs: 21-22, both SEQ ID NOs: 53 and 22, both SEQ ID NOs: 21 and 54, both SEQ ID NOs: 53 and 54, both SEQ ID NOs: 23-24, both SEQ ID NOs: 55 and 24, both SEQ ID NOs: 23 and 54, both SEQ ID NOs: 55 and 56, both SEQ ID NOs: 34-35, both SEQ ID NOs: 61-62, both SEQ ID NOs: 36-37, both SEQ ID NOs: 63-64, both SEQ ID NOs: 40-41, both SEQ ID NOs: 57 and 41, both SEQ ID NOs: 40 and 58, both SEQ ID NOs: 57 and 58, both SEQ ID NOs: 42-43, or both SEQ ID NOs: 65-66. Also, for example, the TCRs, polypeptides, or proteins of the invention can consist essentially of the amino acid sequence(s) of (i) SEQ ID NO: 7, (ii) SEQ ID NO: 51, (iii) SEQ ID NO: 8, (iv) SEQ ID NO: 52, (v) SEQ ID NO: 32, (vi) SEQ ID NO: 33 (vii) SEQ ID NO: 59, or (viii) SEQ ID NO: 60. Furthermore, the TCRs, polypeptides, or proteins of the invention can consist essentially of the following amino acid sequences: (a) any one or more of SEQ ID NOs: 1-6; (b) all of SEQ ID NOs: 1-3; (c) all of SEQ ID NOs: 4-6; or (d) all of SEQ ID NOs: 1-6.

[0080] The TCRs, polypeptides, and proteins of the invention may be of any length, namely, may contain any number of amino acids, so long as the TCRs, polypeptides, or proteins retain their biological activity, such as the ability to specifically bind to the G12D RAS; detect cancer in a mammal; or treat or prevent cancer in a mammal, etc. For example, the length of a polypeptide may be in the range of about 50 to about 5,000 amino acids, such as about 50, about 70, about 75, about 100, about 125, about 150, about 175, about 200, about 300, about 400, about 500, about 600, about 700, about 800, about 900, about 1000 or more amino acids in length. In this regard, the polypeptides of the invention also include oligopeptides.

[0081] The TCRs, polypeptides and proteins of the invention may comprise synthetic amino acids in place of one or more naturally occurring amino acids. Such synthetic amino acids are known in the art and include, for example, aminocyclohexanecarboxylic acid, norleucine, α-amino-n-decanoic acid, homoserine, S-acetylaminomethylcysteine, trans-3- and trans-4-hydroxyproline, 4-aminophenylalanine, 4-nitrophenylalanine, 4-chlorophenylalanine, 4-carboxyphenylalanine, β-phenylserine, β-hydroxyphenylalanine, phenylglycine, α-naphthylalanine, cyclohexylalanine, cyclohexylglycine, indoline-2-carboxylic acid, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, aminomalonic acid, aminomalonic acid monoamide, N'-benzyl-N'-methyllysine, N',N'-dibenzyllysine, 6-hydroxylysine, ornithine, α-aminocyclopentanecarboxylic acid, α-aminocyclohexanecarboxylic acid, α-aminocycloheptanecarboxylic acid, α-(2-amino-2-norbornane)-carboxylic acid, α,γ-diaminobutyric acid, α,β-diaminopropionic acid, homophenylalanine and α-tert-butylglycine.

[0082] The TCRs, polypeptides and proteins of the invention may, for example, be glycosylated, amidated, carboxylated, phosphorylated, esterified, N-acylated, cyclized via, for example, a disulfide bridge, or converted into an acid addition salt and/or optionally dimerized or polymerized or conjugated.

[0083] The TCR, polypeptide, and/or protein of the invention can be produced by methods known in the art, such as, for example, de novo synthesis. In addition, the polypeptides and proteins can be recombinantly produced using the nucleic acids described herein using standard genetic engineering techniques. See, for example, Green and Sambrook, Molecular Cloning: A Laboratory Manual, ^4th ed., Cold Spring Harbor Press, Cold Spring Harbor, NY (2012). Alternatively, the TCR, polypeptides, and/or proteins described herein can be commercially synthesized by any of a variety of commercial organizations. In this regard, the TCR, polypeptides, and proteins of the invention can be synthetic, recombinant, isolated, and/or purified. According to one embodiment, the invention provides an isolated or purified TCR, polypeptide, or protein encoded by any of the nucleic acids or vectors described herein, relative to other aspects of the invention. According to another embodiment of the invention, there is provided an isolated or purified TCR, polypeptide, or protein obtained by expressing any of the nucleic acids or vectors described herein in relation to other aspects of the invention in a cell. According to another embodiment of the invention, there is provided a method for producing any of the TCRs, polypeptides, or proteins described herein, wherein the method comprises culturing any of the host cells or populations of host cells described herein such that the TCR, polypeptide, or protein is obtained.

[0084] The invention includes conjugates, such as bioconjugates, comprising any of the TCRs, polypeptides or proteins (including any of their functional portions or variants), nucleic acids, recombinant expression vectors, host cells, host cell populations, or antibodies or antigen-binding portions thereof, of the invention. Conjugates, as well as methods for synthesizing conjugates, are generally known in the art.

[0085] According to one embodiment of the invention, there is provided a nucleic acid comprising a nucleotide sequence encoding any of the TCRs, polypeptides, or proteins described herein. The term "nucleic acid," as used herein, includes "polynucleotide," "oligonucleotide," and "nucleic acid molecule," and generally refers to a DNA or RNA polymer that may be single-stranded or double-stranded, that may contain natural, non-natural, or modified nucleotides, and that may contain a natural, non-natural, or modified internucleotide linkage, such as a phosphoramidate linkage or a phosphorothioate linkage, in place of the phosphodiester found between nucleotides of an unmodified oligonucleotide. In one embodiment, the nucleic acid comprises complementary DNA (cDNA). It is generally preferred that the nucleic acid does not contain any insertions, deletions, inversions, and/or substitutions. However, it may be appropriate in some examples, as described herein, for the nucleic acid to contain one or more insertions, deletions, inversions and/or substitutions.

[0086] Preferably, the nucleic acids of the invention are recombinant. As used herein, the term "recombinant" refers to (i) molecules that are constructed outside of living cells by joining segments of natural or synthetic nucleic acids to nucleic acid molecules that can replicate in a living cell, or (ii) molecules that are obtained by replication of the molecules described in (i) above. For the purposes of this document, replication may be in vitro replication or in vivo replication.

[0087] In one embodiment of the invention, the nucleic acid comprises the nucleotide sequence of (i) SEQ ID NO: 44 (the nucleotide sequence encoding the variable region of the α chain of the TCR), (ii) the nucleotide sequence of SEQ ID NO: 45 encoding the variable region of the β chain of the TCR, or (iii) both of SEQ ID NO: 44-45.

[0088] Nucleic acids can be engineered based on chemical synthesis and/or enzymatic ligation reactions using methods known herein. See, e.g., Green and Sambrook et al. supra. For example, a nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to enhance the biological stability of the molecules or to increase the physical stability of the duplex formed upon hybridization (e.g., phosphorothioate derivatives and acridine-substituted nucleotides). Examples of modified nucleotides that can be used to form nucleic acids include, but are not limited to, 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxymethyl)uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, β-D-galactosylqueosin, inosine, N ⁶ -isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N ⁶ -substituted adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, β-D-mannosyl queosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N ⁶ -isopentenyladenine, uracil-5-hydroxyacetic acid (v), vibutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-hydroxyacetic acid methyl ester, 3-(3-amino-3-N-2-carboxypropyl) uracil, and 2,6-diaminopurine. Alternatively, one or more of the nucleic acids of the invention can be purchased from any of a variety of commercial organizations.

[0089] The nucleic acid may comprise any nucleotide sequence that encodes any of the TCRs, polypeptides, or proteins described herein. In one embodiment of the invention, the nucleic acid comprises a codon-optimized nucleotide sequence encoding any of the TCRs, polypeptides, or proteins described herein. Without being bound by any particular theory or mechanism, it is believed that codon optimization of the nucleotide sequence increases the translation efficiency of mRNA transcripts. Codon optimization of the nucleotide sequence may include replacing a native codon with another codon that encodes the same amino acid, but can be translated by tRNA that is more readily available in the cell, thereby increasing translation efficiency. Optimization of the nucleotide sequence may also reduce secondary structures of mRNA that would interfere with translation, thereby increasing translation efficiency.

[0090] The invention also provides a nucleic acid comprising a nucleotide sequence that is complementary to the nucleotide sequence of any of the nucleic acids described herein, or a nucleotide sequence that hybridizes under stringent conditions to the nucleotide sequence of any of the nucleic acids described herein.

[0091] A nucleotide sequence that hybridizes under stringent conditions preferentially hybridizes under high stringency conditions. By "high stringency conditions" is meant that the nucleotide sequence specifically hybridizes to a target sequence (the nucleotide sequence of any of the nucleic acids described herein) in an amount that is noticeably stronger than non-specific hybridization. High stringency conditions include conditions that will distinguish a polynucleotide with an exact complementary sequence or a polynucleotide containing only a few scattered mismatches from a random sequence that happens to have several small regions (e.g., 3-10 bases) that match the nucleotide sequence. Such small regions of complementarity are more easily melted than a full-length complement of 14-17 or more bases, and hybridization under high stringency conditions makes them easily distinguishable. Relatively high-stringency conditions will include, for example, low-salt and/or high-temperature conditions, such as those provided with approximately 0.02-0.1 M NaCl or equivalent, at temperatures of approximately 50-70°C. Such high-stringency conditions allow for few, if any, mismatches between the nucleotide sequence and the template or target strand and are particularly suitable for detecting the expression of any of the TCRs of the invention. It will generally be appreciated that conditions can be made more stringent by adding increasing amounts of formamide.

[0092] The invention also provides a nucleic acid comprising a nucleotide sequence that is at least about 70% or more, such as about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to any of the nucleic acids described herein. In this regard, the nucleic acid may consist essentially of any of the nucleotide sequences described herein.

[0093] According to one embodiment of the invention, there is provided an isolated or purified nucleic acid comprising, from 5' to 3', a first nucleic acid sequence and a second nucleotide sequence, wherein the first and second nucleotide sequences, respectively, encode the amino acid sequences of SEQ ID NOs: 7 and 8; 51 and 8; 7 and 52; 51 and 52; 8 and 7; 8 and 51; 52 and 7; 52 and 51; 21 and 22; 21 and 54; 53 and 22; 53 and 54; 22 and 21; 54 and 21; 22 and 53; 54 and 53; 23 and 24; 55 and 24; 23 and 56; 55 and 56; 24 and 23; 24 and 55; 56 and 55; 56 and 23; 32 and 33; 33 and 32; 59 and 60; 60 and 59; 34 and 35; 35 and 34; 61 and 62; 62 and 61; 36 and 37; 37 and 36; 63 and 64; 64 and 63; 40 and 41; 57 and 41; 40 and 58; 57 and 58; 41 and 40; 41 and 57; 58 and 40; 58 and 57; 42 and 43; 43 and 42; 65 and 66; or 66 and 65.

[0094] In one embodiment of the invention, the isolated or purified nucleic acid further comprises a third nucleotide sequence located between the first and second nucleotide sequences, wherein the third nucleotide sequence encodes a cleavable linker peptide. In an embodiment of the invention, the cleavable linker peptide comprises the amino acid sequence RAKRSGSGATNFSLLKQAGDVEENPGP (SEQ ID NO: 25).

[0095] The nucleic acids of the invention can be included in a recombinant expression vector. In this regard, the invention provides a recombinant expression vector comprising any of the nucleic acids of the invention. In one embodiment of the invention, the recombinant expression vector comprises a nucleotide sequence encoding an α chain, a β chain, and a linker peptide.

[0096] For the purposes of this document, the term "recombinant expression vector" refers to a genetically modified oligonucleotide or polynucleotide construct that allows for the expression of an mRNA, protein, polypeptide, or peptide by a host cell, wherein the construct comprises a nucleotide sequence encoding the mRNA, protein, polypeptide, or peptide, and the vector is contacted with the cell under conditions sufficient to express the mRNA, protein, polypeptide, or peptide in the cell. The vectors of the invention do not occur in nature as a whole. However, parts of the vectors may occur in nature. The recombinant expression vectors of the invention may contain any type of nucleotide, including, but not limited to, DNA and RNA, which may be single-stranded or double-stranded, synthesized or obtained in part from natural sources, and which may contain natural, non-natural, or altered nucleotides. Recombinant expression vectors may contain naturally occurring, non-naturally occurring internucleotide linkages, or both. Preferably, non-naturally occurring or altered nucleotides or internucleotide linkages do not interfere with vector transcription or replication.

[0097] The recombinant expression vector of the invention may be any suitable recombinant expression vector and may be used to transform or transfect any suitable host cell. Suitable vectors include vectors designed for production and propagation or for expression, or both, such as plasmids and viruses. The vector may be selected from the pUC series (Fermentas Life Sciences), the pBluescript series (Stratagene, LaJolla, CA), the pET series (Novagen, Madison, WT), the pGEX series (Pharmacia Biotech, Uppsala, Sweden), and the pEX series (Clontech, Palo Alto, CA). Bacteriophage-based vectors such as λGT10, λGT11, λZap11 (Stratagene), λEMBL4, and λNM1149 may also be used. Examples of plant expression vectors include pBI01, pBI101.2, pBI101.3, pBI121, and pBIN19 (Clontech). Examples of animal expression vectors include pEUK-CI, pMAM, and pMAMneo (Clontech). Preferably, the recombinant expression vector is a viral vector, such as a retroviral vector. In a particularly preferred embodiment, the recombinant expression vector is the MSGV1 vector. In one embodiment of the invention, the recombinant expression vector is a transposon or a lentiviral vector.

[0098] Recombinant expression vectors of the invention can be prepared using standard genetic engineering techniques, such as those described in Green and Sambrook et al., supra. Expression vector constructs, which are circular or linear, can be prepared to contain a replication system that is functional in a prokaryotic or eukaryotic host cell. Replication systems can be derived from, for example, ColEI, 2 μ plasmidase, λ, SV40, bovine papillomavirus, and the like.

[0099] It is desirable that the recombinant expression vector contain regulatory sequences, such as transcription and translation start and stop codons, that are specific to the type of host cell (e.g., bacterium, fungus, plant, or animal) into which the vector is to be introduced, depending on the particular case and taking into account whether the vector is DNA or RNA based.

[0100] The recombinant expression vector may include one or more marker genes that allow selection of transformed or transfected host cells. Marker genes include biocide resistance, such as resistance to antibiotics, heavy metals, etc., complementation in an auxotrophic host cell to ensure prototrophy, etc. Suitable marker genes for expression vectors of the invention include, for example, neomycin/G418 resistance genes, hygromycin resistance genes, histidinol resistance genes, tetracycline resistance genes, and ampicillin resistance genes.

[0101] A recombinant expression vector may comprise a native or non-native promoter operably linked to a nucleotide sequence encoding a TCR, polypeptide, or protein, or to a nucleotide sequence that is complementary to or that hybridizes with a nucleotide sequence encoding a TCR, polypeptide, or protein. The selection of promoters, such as strong, weak, inducible, tissue-specific, and developmental stage-specific, is within the skill of one of ordinary skill in the art. Similarly, the association of a nucleotide sequence with a promoter is also within the skill of one of ordinary skill in the art. The promoter may be a non-viral promoter or a viral promoter, such as the cytomegalovirus (CMV) promoter, the SV40 promoter, the RSV promoter, and the promoter found in the mouse stem cell virus long terminal repeat.

[0102] Recombinant expression vectors of the invention can be designed for either transient expression or stable expression, or both. Furthermore, recombinant expression vectors can be made for constitutive expression or inducible expression.

[0103] Furthermore, recombinant expression vectors can be made to include a suicide gene. As used herein, the term "suicide gene" refers to a gene that causes the death of a cell expressing the suicide gene. The suicide gene can be a gene that confers sensitivity to an agent, such as a drug, on a cell expressing the gene and causes the cell to die when the cell contacts or is exposed to the agent. Suicide genes are known in the art and include, for example, the herpes simplex virus (HSV) thymidine kinase (TK) gene, the cytosine deaminase gene system, the purine nucleoside phosphorylase gene system, the nitroreductase gene system, and the inducible caspase 9 gene system.

[0104] According to another embodiment of the invention, there is further provided a host cell comprising any of the recombinant expression vectors described herein. As used herein, the term "host cell" refers to any type of cell that can contain a recombinant expression vector of the invention. The host cell may be a eukaryotic cell, such as a plant, animal, fungal, or algal cell, or may be a prokaryotic cell, such as a bacterium or protozoa. The host cell may be a cultured cell or a primary cell, i.e., isolated directly from an organism, such as a human or a mouse. The host cell may be an adherent cell or a suspension cell, i.e., a cell that grows in suspension. Suitable host cells are known in the art and include, for example, E. coli DH5a cells, Chinese hamster ovary cells, monkey VERO cells, COS cells, HEK293 cells, and the like. For the purpose of amplifying or replicating a recombinant expression vector, the host cell is preferably a prokaryotic cell, such as a DH5α cell. For the purpose of producing a recombinant TCR, polypeptide, or protein, the host cell is preferably a mammalian cell. Most preferably, the host cell is a human cell. While the host cell can be any cell type, can be from any tissue type, and can be at any stage of development, the host cell is preferably a peripheral blood lymphocyte (PBL) or a peripheral blood mononuclear cell (PBMC). More preferably, the host cell is a T cell. In one embodiment of the invention, the host cell is a human lymphocyte. In one embodiment of the invention, the host cell is selected from the group consisting of a T cell, a natural killer T cell (NKT), an invariant natural killer T cell (iNKT), and a natural killer (NK). According to another embodiment of the invention, a method for producing a host cell expressing a TCR that has antigen specificity for a peptide is provided. (SEQ ID NO: 29), wherein the method comprises contacting a cell with any of the vectors described herein under conditions that allow the vector to be introduced into the cell.

[0105] For the purposes of this document, the T cell may be any T cell, such as a cultured T cell, for example, a primary T cell or a T cell from a cultured T cell line, for example, Jurkat, SupT1, etc., or a T cell obtained from a mammal. If obtained from a mammal, the T cell may be obtained from many sources, including, but not limited to, blood, bone marrow, lymph node, thymus, or other tissues or fluids. T cells may also be enriched or purified. Preferably, the T cell is a human T cell. The T cell may be any type of T cell and may be at any stage of development, including, but not limited to, CD4 ⁺ /CD8 ⁺ double-positive T cells, CD4 ⁺ T helper cells such as _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.

[0106] Also provided according to the invention is a population of cells comprising at least one host cell described herein. The population of cells can be a heterogeneous population comprising a host cell comprising any of the described recombinant expression vectors, in addition to at least one other cell, for example, a host cell (e.g., a T cell) that does not comprise any of the recombinant expression vectors, or a cell other than a T cell, for example, a B cell, a macrophage, a neutrophil, a erythrocyte, a hepatocyte, an endothelial cell, an epithelial cell, a muscle cell, a brain cell, etc. Alternatively, the population of cells can be a substantially homogeneous population, wherein the population comprises primarily host cells (e.g., consisting essentially of) comprising a recombinant expression vector. The population may also be a clonal population of cells, in which all cells in the population are clones of a single host cell containing the recombinant expression vector, such that all cells in the population contain the recombinant expression vector. In one embodiment of the invention, the population of cells is a clonal population comprising host cells containing the recombinant expression vector, as described herein.

[0107] In one embodiment of the invention, the number of cells in a population can be rapidly expanded. The expansion of the number of T cells can be accomplished by any of a variety of methods known in the art, as described, for example, in U.S. Patent 8,034,334; U.S. Patent 8,383,099; U.S. Patent Application Publication No. 2012/0244133; Dudley et al., J. Immunother., 26:332-42 (2003); and Riddell et al., J. Immunol. Methods, 128:189-201 (1990). In one embodiment, the expansion of the number of T cells is accomplished by culturing the T cells with OKT3 antibody, IL-2, and feeder PBMCs (e.g., irradiated allogeneic PBMCs).

[0108] TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, and host cells (including populations thereof) of the invention may be isolated and/or purified. The term "isolated," as used herein, means removed from its natural environment. The term "purified," as used herein, means with increased purity, where "purity" is a relative term and is not necessarily understood as absolute purity. For example, purity may be at least about 50%, may be greater than about 60%, about 70%, about 80%, about 90%, about 95%, or may be about 100%.

[0109] The TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, and host cells (including populations thereof) of the invention, all of which are hereinafter collectively referred to as "TCR materials of the invention," can be formulated in a composition, such as a pharmaceutical composition. Accordingly, the invention provides a pharmaceutical composition comprising any of the TCRs, polypeptides, proteins, nucleic acids, expression vectors, and host cells (including populations thereof) described herein and a pharmaceutically acceptable carrier. Pharmaceutical compositions of the invention comprising any of the TCR materials of the invention may comprise more than one TCR material of the invention, such as a polypeptide and a nucleic acid, or two or more different TCRs. Alternatively, the pharmaceutical composition may comprise the TCR material of the invention in combination with other pharmaceutically active agent(s) or drug(s), such as a chemotherapeutic agent, for example, asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, vincristine, etc.

[0110] Preferably, the carrier is a pharmaceutically acceptable carrier. With respect to pharmaceutical compositions, the carrier may be any of the carriers conventionally used for the particular TCR material of the invention under consideration. Methods for preparing the administered compositions are known or obvious to those skilled in the art and are described in more detail, for example, in Remington: The Science and Practice of Pharmacy, ^22nd Ed., Pharmaceutical Press (2012). Preferably, the pharmaceutically acceptable carrier is one that does not have deleterious side effects or toxicity under the conditions of use.

[0111] The choice of carrier will be determined in part by the particular TCR material of the invention, as well as the particular route used to administer the TCR material of the invention. Accordingly, there are a variety of suitable formulations of the pharmaceutical composition of the invention. Suitable formulations may include any of those for parenteral, subcutaneous, intravenous, intramuscular, intraarterial, intrathecal, intratumoral, or intraperitoneal administration. More than one route may be used to administer the TCR materials of the invention, and, in certain examples, a particular route may provide a more immediate and more effective response than another route.

[0112] Preferably, the TCR material of the invention is administered by injection, such as intravenously. When the TCR material of the invention is a host cell (or population thereof) expressing the TCR of the invention, the pharmaceutically acceptable carrier for the cells for injection may include any isotonic carrier, such as, for example, normal saline (about 0.90% w/v NaCl in water, about 300 mOsm/L NaCl in water, or about 9.0 g NaCl per liter of water), NORMOSOL R electrolyte solution (Abbott, Chicago, IL), PLASMA-LYTE A (Baxter, Deerfield, IL), about 5% dextrose in water, or Ringer's lactate. In one embodiment, the pharmaceutically acceptable carrier is supplemented with human serum albumin.

[0113] For the purposes of the invention, the amount or dose (e.g., the number of cells when the TCR material of the invention is one or more cells) of the TCR material of the invention administered should be sufficient to produce an effect, e.g., a therapeutic or prophylactic response, in a subject or animal within a reasonable period of time. For example, the dose of the TCR material of the invention should be sufficient to bind to a cancer antigen (e.g., G12D RAS) or detect, treat, or prevent cancer within a period of about 2 hours or more, such as 12 to 24 hours or more, from the time of administration. In specific embodiments, the time period could be even longer. The dose will be determined by the potency of the particular TCR material of the invention and the condition of the animal (e.g., a human), as well as the body weight of the animal (e.g., a human) being treated.

[0114] Many assays for determining the dose to be administered are known in the art. For purposes of the invention, an assay that involves comparing the extent to which target cells are lysed or IFN-γ is secreted by T cells expressing the TCR, polypeptide, or protein of the invention when a given dose of such T cells is administered to a mammal across a variety of mammals, each of which is given a different dose of T cells, could be used to determine the initial dose to be administered to the mammal. The extent to which target cells are lysed or IFN-γ is secreted when a given dose is administered can be analyzed by methods known in the art.

[0115] The dosage of the TCR material of the invention will also be determined by the existence, nature, and extent of any adverse side effects that may accompany the administration of the particular TCR material of the invention. Typically, the treating physician will decide on the dosage of the TCR material of the invention with which to treat each individual patient, taking into account a variety of factors, such as age, body weight, general health, diet, gender, the TCR material of the invention to be administered, the route of administration, and the severity of the cancer to be treated. In one embodiment, in which the TCR material of the invention is a population of cells, the number of cells administered per infusion can vary, for example, from about 1 x 10 ⁶ to about 1 x 10 ¹² cells or more. In certain embodiments, less than 1 x 10 ⁶ cells can be administered.

[0116] One of ordinary skill in the art will readily appreciate that the TCR materials of the invention can be modified in a variety of ways such that the therapeutic or prophylactic efficacy of the TCR materials of the invention is enhanced by the modification. For example, the TCR materials of the invention can be conjugated either directly or indirectly through a bridge to a chemotherapeutic agent. The practical application of conjugating compounds to a chemotherapeutic agent is known in the art. One of ordinary skill in the art will recognize that sites on the TCR materials of the invention that are not necessary for the function of the TCR materials of the invention are sites suitable for attachment of a bridge and/or a chemotherapeutic agent, provided that the bridge and/or chemotherapeutic agent, when attached to the TCR materials of the invention, does not interfere with the function of the TCR materials of the invention, i.e., the ability to bind to G12D RAS or to detect, treat, or prevent cancer.

[0117] It is contemplated that the pharmaceutical compositions, TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, and cell populations of the invention can be used in methods for treating or preventing cancer. Without being bound by a particular theory, it is believed that the TCRs of the invention specifically bind to G12D RAS such that the TCR (or a related polypeptide or protein of the invention), when expressed by a cell, is capable of mediating an immune response to a target cell expressing G12D RAS. In this regard, according to one embodiment of the invention, there is provided a method for treating or preventing cancer in a mammal, comprising administering to the mammal any of the pharmaceutical compositions, TCRs, polypeptides or proteins described herein, any nucleic acid or recombinant expression vector comprising a nucleotide sequence encoding any of the TCRs, polypeptides, proteins described herein, or any host cell or population of cells comprising a recombinant vector that encodes any of the TCRs, polypeptides or proteins described herein, in an amount effective for treating or preventing cancer in the mammal.

[0118] According to one embodiment of the invention, there is provided a method for inducing an immune response to a cancer in a mammal, comprising administering to the mammal any of the pharmaceutical compositions, TCRs, polypeptides, or proteins described herein, any nucleic acid or recombinant expression vector comprising a nucleotide sequence encoding any of the TCRs, polypeptides, proteins described herein, or any host cell or population of cells comprising a recombinant vector that encodes any of the TCRs, polypeptides, or proteins described herein, in an amount effective to induce an immune response to a cancer in the mammal.

[0119] According to one embodiment of the invention, there is provided any of the pharmaceutical compositions, TCRs, polypeptides, or proteins described herein, any nucleic acid or recombinant expression vector comprising a nucleotide sequence encoding any of the TCRs, polypeptides, proteins described herein, or any host cell or population of cells comprising a recombinant vector that encodes any of the TCRs, polypeptides, or proteins described herein, for use in the treatment or prevention of cancer in a mammal.

[0120] According to one embodiment of the invention, there is provided any of the pharmaceutical compositions, TCRs, polypeptides, or proteins described herein, any nucleic acid or recombinant expression vector comprising a nucleotide sequence encoding any of the TCRs, polypeptides, or proteins described herein, or any host cell or population of cells comprising a recombinant vector that encodes any of the TCRs, polypeptides, or proteins described herein, for use in inducing an immune response to cancer in a mammal.

[0121] The terms "treat" and "prevent," and words derived from them, as used herein, do not necessarily imply a 100% or complete cure or prevention. Rather, there are varying degrees of treatment or prevention that one of ordinary skill in the art would recognize as providing potential benefit or therapeutic effect. Accordingly, the methods of the invention may provide any number of any level of treatment or prevention of cancer in a mammal. Furthermore, the treatment or prevention provided by the method of the invention may include treating or preventing one or more conditions or symptoms of the cancer being treated or prevented. For example, treatment or prevention may include promoting tumor regression. Furthermore, for the purposes of this document, the term "prevention" may encompass delaying the onset of cancer or a symptom or condition thereof. Alternatively or additionally, the term "prevention" may encompass preventing or delaying the recurrence of cancer or a symptom or condition thereof.

[0122] Also according to an embodiment of the invention, a method for detecting the presence of cancer in a mammal is provided. The method comprises (i) contacting a sample containing one or more cells from the mammal with any of the TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, cell populations, or pharmaceutical compositions of the invention described herein, thereby forming a complex, and (ii) detecting the complex, wherein detection of the complex indicates the presence of cancer in the mammal.

[0123] With respect to the method for detecting cancer in a mammal according to the invention, the cell sample may be a sample containing whole cells, lysates thereof, or a fraction of whole cell lysates, such as a nuclear or cytoplasmic fraction, a total protein fraction, or a nucleic acid fraction.

[0124] For the purposes of the method for detecting cancer according to the invention, contacting may occur in vitro or in vivo with respect to a mammal. Preferably, contacting occurs in vitro.

[0125] Furthermore, detection of the complex can occur through any number of routes known in the art. For example, the TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, or cell populations of the invention described herein can be labeled with a detectable label, such as, for example, a radioisotope, a fluorophore (e.g., fluorescein isothiocyanate (FITC), phycoerythrin (PE)), an enzyme (e.g., alkaline phosphatase, horseradish peroxidase), and elemental particles (e.g., gold particles).

[0126] For the purposes of the methods of the invention in which host cells or cell populations are administered, the cells may be cells that are allogeneic or autologous with respect to the mammal. Preferably, the cells are autologous with respect to the mammal.

[0127] With respect to the methods of the invention, the cancer can be any cancer, including, for example, any of the following cancers: acute lymphocytic cancer, acute myeloid leukemia, alveolar rhabdomyosarcoma, bone cancer, brain cancer, breast cancer, cancer of the anus, anal canal, or rectum, eye cancer, intrahepatic bile duct cancer, joint cancer, cancer of the neck, gallbladder, or pleura, cancer of the nose, nasal cavity, or middle ear, oral cancer, vaginal cancer, vulvar cancer, chronic lymphocytic leukemia, chronic myeloid cancer, colon cancer, colorectal cancer, endometrial cancer, esophageal cancer, cervical cancer, gastrointestinal carcinoid tumor, glioma, Hodgkin's lymphoma, hypopharyngeal cancer, kidney 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, omental cancer, mesenteric cancer, pharyngeal cancer, prostate cancer, rectal cancer, kidney cancer, skin cancer, small bowel cancer, soft tissue cancer, stomach cancer, testicular cancer, thyroid cancer, uterine cancer, ureteral cancer, and bladder cancer. Preferred cancers are pancreatic cancer, colorectal cancer, lung cancer, endometrial cancer, ovarian cancer, or prostate cancer. Preferably, lung cancer is lung adenocarcinoma, ovarian cancer is ovarian epithelial cancer, and pancreatic cancer is pancreatic adenocarcinoma. In one embodiment of the invention, the amino acid sequence of mutant human RAS with a glycine substitution at position 12 for aspartic acid is expressed in cancer, wherein the amino acid sequence of mutant human RAS is the amino acid sequence of mutant human KRAS, mutant human HRAS, or mutant human NRAS, and wherein position 12 is determined based on human KRAS WT, human HRAS WT, or human NRAS WT, respectively. Mutant human KRAS, mutant human HRAS, and mutant human NRAS expressed in cancer may be as defined herein in relation to other aspects of the invention.

[0128] The mammal referred to in the methods of the invention may be any mammal. As used herein, the term "mammal" refers to any mammal, including, but not limited to, mammals of the order Rodentia, such as mice and hamsters, and mammals of the order Logomorpha, such as rabbits. Preferably, the mammals are from the order Carnivora, including Felines (cats) and Canines (dogs). More preferably, the mammals are from the order Artiodactyla, including Bovines (cows) and Swines (pigs), or from the order Perssodactyla, including Equines (horses). Most preferably, the mammals belong to the order Primates, Ceboids, or Simoids (monkeys), or to the order Anthropoids (humans and great apes). The most preferred mammal is man.

[0129] It will be noted that the above represents only exemplary embodiments. Other illustrative embodiments will be apparent from the complete description herein. It will also be understood by one skilled in the art that each of these embodiments can be used in various combinations with other embodiments provided herein.

[0130] The following examples further illustrate the invention, but should, of course, in no way be considered as limiting its scope.

Example 1

[0131] This example demonstrates the isolation of a TCR with antigen specificity for human KRAS with the G12D mutation.

[0132] Patient 4373's endometrial cancer progressed after treatment with autologous PBL transduced with a mouse TCR with antigen specificity for HLA-A11-restricted human RAS G12D. The patient's TILs were screened for reactivity to KRAS G12D as described below. TILs from tumor fragments F4, F5, F6, F8, F9, and F10 were co-cultured with the following target cells:

- 4373 autologous DC mRNA transfected with a tandem minigene (TMG) encoding the wild-type KRAS (WT) TMG peptide:

- 4373 autologous DC mRNA transfected with TMG encoding the mutant KRAS peptide (Mut):

- long peptide G12 WT KRAS (LP) (SEQ ID NO: 27);

- G12D Mut KRAS LP (SEQ ID NO: 26); or

- minimal epitope of KRAS (ME) A11 (G12D 9mer+10mer) + (SEQ ID NO: 28 and 29).

[0133] As controls, transduced cells were cultured alone (TILs only) or co-cultured with dimethyl sulfoxide (DMSO) or anti-CD28/anti-COD magnetic bead material.

[0134] Interferon-gamma (IFN-γ) secretion after co-culture was measured by enzyme-linked immunosorbent assay (ELISpot). The results are shown in Fig. 1A. The percentage of cells expressing 4-1 BB and OX40 was measured by flow cytometry analysis. The results are shown in Fig. 1B. As shown in Figs. 1A-1B, TILs with anti-G12D reactivity were detected in the F8 tumor fragment.

[0135] TILs from the F8 tumor fragment were separated into single-cell samples. TCR with antigen specificity for human KRAS with the G12D mutation presented by HLA-A11 was isolated from the TILs. To determine the sequence of the reactive 4373 TCR, reactive TILs were sorted by fluorescence-activated cell sorting (FACS) based on the upregulation of the T cell activation marker 4-1BB. The cells were then lysed, and the TCR transcripts were sequenced by Sanger. The amino acid sequences of the variable regions of the ai 4373 TCR chains are shown in Table 5. CDRs are underlined.

Example 2

[0136] This example demonstrates a method for producing a retroviral vector containing a nucleotide sequence encoding the human anti-G12D TCR of Example 1 with modified murine constant regions.

[0137] A nucleic acid sequence encoding the human G12D RAS-reactive 4373 TCR of Example 1 and comprising an LVL-modified mouse constant region with a cysteine substitution was cloned into a retroviral expression vector. The mouse α chain constant region comprised the amino acid sequence of SEQ ID NO: 17, wherein X at position 48 is Cys, X at position 112 is Leu, X at position 114 is Ile, and X at position 115 is Val (SEQ ID NO: 38). The resulting full-length α chain comprised the amino acid sequence of SEQ ID NO: 40. The β chain constant region comprised the amino acid sequence of SEQ ID NO: 18, wherein X at position 57 is Cys (SEQ ID NO: 39). The resulting full-length β chain contained the amino acid sequence of SEQ ID NO: 41. The linker containing the amino acid sequence (SEQ ID NO: 25), was located between the constant region of the α chain and the variable region of the β chain. The vector contained an expression cassette containing the nucleotide sequence of SEQ ID NO: 46 (codon-optimized nucleotide sequence encoding, from 5' to 3': TCR β chain, linker, TCR α chain), which encoded the amino acid sequence of SEQ ID NO: 47 (amino acid sequence comprising, from N-terminus to C-terminus, TCR β chain, linker, TCR α chain).

Example 3

[0138] This example demonstrates that the anti-G12D TCR of Example 2 (with human variable regions) provides the same or better reactivity as the mouse anti-G12D TCR of Example 1.

[0139] CD8+ autologous PBLs from patient 4373 were transduced with a retroviral expression vector encoding (i) the G12D RAS-reactive 4373 TCR of Example 2 (with human variable regions) (also referred to herein as "TCR2"), (ii) a second TCR (referred to herein as "TCR1"), which was also obtained by single-cell sequencing of the reactive TILs shown in Fig. 1, or (iii) the HLA-A11-restricted, murine anti-KRAS G12D TCR of Example 1 (control). The reactivity of the CD8+ transduced cells was tested after co-culture with the following target cells:

- COS HLA-A2 transduced cells,

- cell line COS HLA-A2-G12 WT KRAS,

- COS HLA-A11 transduced cells,

- cell line COS HLA-A11-G12D or

- T cells only (no target cells) (-).

[0140] The percentage of cells expressing 4-1 BB and OX40 was measured after co-culture with target cells. The results are shown in Fig. 2. Transduced cells were also subjected to titration experiments for a minimal HLA-A11 epitope.

[0141] In a separate experiment, autologous CD8+ PBL from Patient 4373 were transduced with a retroviral expression vector encoding (i) the G12D RAS-reactive 4373 TCR of Example 2 (with human variable regions), (ii) TCR1, (iii) the HLA-A11-restricted, murine anti-KRAS G12D TCR of Example 1, or (iv) GFP. Cells transduced with empty vector ("Moek Td" (Td stands for Transduction)) served as an additional control. The reactivity of CD8+ transduced cells was analyzed after co-culture with the following target cells:

- autologous dendritic cells (DC) transduced with the full-length (FL) WT KRAS gene;

- autologous DC transduced with the FL KRAS gene with the G12D mutation;

- autologous DC transduced with WT KRAS LP (SEQ ID NO: 27);

- autologous DC transduced with G12D Mut KRAS LP (SEQ ID NO: 26);

- minimal epitope of KRAS (ME) A02 WT;

- ME A02 G12D (SEQ ID NO: 50);

- a mixture of ME HLA-A11 WT (a mixture of peptides WT 9-mer SEQ ID NO: 30 and WT 10-MER SEQ ID NO: 31); or

- mixture of ME HLA-A11 G12D (mixture of peptides G12D 9-mer SEQ ID NO: 28 and G12D 10-MER SEQ ID NO: 29).

[0142] As a control, transduced cells were cultured alone (T cells only) or co-cultured with DMSO or anti-CD28/anti-CD3 magnetic bead material. IFN-γ secretion was measured by ELISpot. The results are shown in Fig. 3A and Table 6. The percentage of cells expressing 4-1BB and OX40 was measured. The results are shown in Fig. 3B.

[0143] In a separate experiment, autologous PBLs from Patient 4373 were transduced with a retroviral expression vector encoding (i) the G12D RAS-reactive 4373 TCR of Example 2 (with human variable regions) or (ii) the HLA-A11-restricted, murine anti-KRAS G12D TCR of Example 1.

[0144] Autologous DCs were loaded with the following peptides at the concentrations shown in Table 7: mutant peptide minimal epitope (ME) with 9 amino acid residues (SEQ ID NO: 28), mutant peptide minimal epitope with 10 amino acid residues (SEQ ID NO: 29), minimal WT epitope peptide with 9 amino acid residues (SEQ ID NO: 30) or a minimal WT epitope peptide with 10 amino acid residues (SEQ ID NO: 31). IFN-γ secretion was measured by ELISpot. The results are shown in Table 7.

[0145] As shown in Fig. 2-3B and Tables 6-7, the anti-G12D TCR of Example 2 (with human variable regions) provided the same or better reactivity compared to the anti-G12D mouse TCR of Example 1.

Example 4

[0146] This example demonstrates that PBLs transduced with the TCR against G12D of Example 2 (with human variable regions) specifically recognize HLA-A11-restricted G12Ds with high avidity.

[0147] Autologous PBLs from Patient 4373 were transduced with a retroviral expression vector encoding the G12D RAS-reactive 4373 TCR of Example 2 (with human variable regions).

[0148] Autologous DCs were loaded with the following peptides at the concentrations shown in Fig. 4A: 10 amino acid residue minimal epitope mutant peptide (SEQ ID NO: 29) or a minimal WT epitope peptide with 10 amino acid residues (SEQ ID NO: 31). IFN-γ secretion was measured by ELISpot. The results are shown in Fig. 4A.

Example 5

[0149] This example demonstrates that CD8+ PBL transduced with the TCR against G12D of Example 2 (with human variable regions) specifically recognizes HLA-A11-restricted G12D with high avidity.

[0150] Autologous CD8+ PBLs from Patient 4373 were transduced with a retroviral expression vector encoding the G12D RAS-reactive 4373 TCR of Example 2 (with human variable regions).

[0151] Autologous DCs were loaded with the following peptides at the concentrations shown in Fig. 4B: mutant peptide minimal epitope with 10 amino acid residues (SEQ ID NO: 29) or a minimal WT epitope peptide with 10 amino acid residues (SEQ ID NO: 31). Expression of 4-1 BB and OX40 was measured by FACS. The results are shown in Fig. 4B.

Example 6

[0152] This example demonstrates that CD4+ PBL transduced with the TCR against G12D of Example 2 (with human variable regions) specifically recognize HLA-A11-restricted G12D with high avidity.

[0153] Autologous CD4+ PBLs from Patient 4373 were transduced with a retroviral expression vector encoding the G12D RAS-reactive 4373 TCR of Example 2 (with human variable regions).

[0154] Autologous DCs were loaded with the following peptides at the concentrations shown in Fig. 4C: mutant peptide minimal epitope with 10 amino acid residues (SEQ ID NO: 29) or a minimal WT epitope peptide with 10 amino acid residues (SEQ ID NO: 31). Expression of 4-1 BB and OX40 was measured by FACS. The results are shown in Fig. 4C.

Example 7

[0155] This example demonstrates that CD8+ PBL transduced with the TCR against G12D of Example 2 (with human variable regions) specifically recognize HLA-A11-restricted G12D with high avidity.

[0156] Autologous CD8+ PBLs from Patient 4373 were transduced with a retroviral expression vector encoding the G12D RAS-reactive 4373 TCR of Example 2 (with human variable regions).

[0157] COS cells were loaded with the following peptides at the concentrations shown in Fig. 5A: mutant peptide minimal epitope with 10 amino acid residues (SEQ ID NO: 29) or a minimal WT epitope peptide with 10 amino acid residues (SEQ ID NO: 31). IFN-γ secretion was measured by ELISpot. The results are shown in Fig. 5A.

Example 8

[0158] This example demonstrates that CD8+ PBL transduced with the TCR against G12D of Example 2 (with human variable regions) specifically recognize HLA-A11-restricted G12D with high avidity.

[0159] Autologous CD8+ PBLs from Patient 4373 were transduced with a retroviral expression vector encoding the G12D RAS-reactive 4373 TCR of Example 2 (with human variable regions).

[0160] COS cells were transduced with HLA-A11 and loaded with the following peptides at the concentrations shown in Fig. 5B: mutant peptide minimal epitope with 10 amino acid residues (SEQ ID NO: 29) or a minimal WT epitope peptide with 10 amino acid residues (SEQ ID NO: 31). Expression of 4-1 BB and OX40 was measured by FACS. The results are shown in Fig. 5B.

Example 9

[0161] This example demonstrates that CD4+ PBL transduced with the TCR against G12D of Example 2 (with human variable regions) specifically recognize HLA-A11-restricted G12D with high avidity.

[0162] Autologous CD4+ PBL from patient 4373 were transduced with a retroviral expression vector encoding the G12D RAS-reactive 4373 TCR of Example 2 (with human variable regions).

[0163] COS cells were loaded with the following peptides at the concentrations shown in Fig. 5C: mutant peptide minimal epitope with 10 amino acid residues (SEQ ID NO: 29) or a minimal WT epitope peptide with 10 amino acid residues (SEQ ID NO: 31). Expression of 4-1 BB and OX40 was measured by FACS. The results are shown in Fig. 5C.

[0164] All references, including publications, patent applications, and patents, cited in this document are herein incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth herein in its entirety.

[0165] The use of the terms "at least one" and "a" and similar references in the context of the description of the invention (especially in the context of the following claims) shall be construed as encompassing both the singular and the plural unless otherwise indicated herein or clearly contradicted by the context. The use of the term "at least one" followed by a listing of one or more items (e.g., "at least one of A and B") shall be construed as meaning a single item selected from the listed items (A or B) or any combination of two or more of the listed items (A or B), unless otherwise indicated herein or clearly contradicted by the context. The terms "comprising," "having," "including," and "comprising" shall be construed as open-ended terms (i.e., meaning "including, but not limited to...") unless otherwise indicated herein. The disclosure of ranges of values in this document is intended only to serve as a shortcut for individually referring to each individual value falling within the range, unless otherwise indicated herein, and each individual value is included in the description of the invention as if it were individually listed herein. All methods described in this document can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of all possible examples or introductory words before an example (e.g., "such as") provided in this document is intended only to better illuminate the invention and does not impose a limitation on the scope of the invention unless otherwise stated. Language in the description of the invention should not be construed as indicating any unclaimed element as essential to the practice of the invention.

[0166] Preferred embodiments of the present invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations from these preferred embodiments may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect that those skilled in the art will employ such variations where appropriate, and the inventors intend to practice the present invention otherwise than as specifically described herein. Accordingly, the present invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto, as permitted by applicable law. Furthermore, any combination of the above-described elements in all their possible variations is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

--->

SEQUENCE LIST

<110> DZE UNITED STATES OF AMERICA, EZ REPRESENTED BY DZE

SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES

<120> HLA CLASS I RESTRICTED T-CELL RECEPTORS AGAINST

RAS WITH G12D MUTATION

<130> 751506

<150> US 62/975,544

<151> 2020-02-12

<160> 67

<170> PatentIn version 3.5

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Thr Phe Trp His Asn Pro Arg Asn His Phe Arg Cys Gln Val Gln Phe

85 90 95

His Gly Leu Ser Glu Glu Asp Lys Trp Pro Glu Gly Ser Pro Lys Pro

100 105 110

Val Thr Gln Asn Ile Ser Ala Glu Ala Trp Gly Arg Ala Asp Cys Gly

115 120 125

Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val Leu Ser Ala Thr Ile Leu

130 135 140

Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu Tyr Ala Val Leu Val Ser

145 150 155 160

Thr Leu Val Val Met Ala Met Val Lys Arg Lys Asn Ser

165 170

<210> 21

<211> 282

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> MISC_FEATURE

<222> (193) .. (193)

<223> Xaa is Thr or Cys

<220>

<221> MISC_FEATURE

<222> (257)..(257)

<223> Xaa represents Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp

<220>

<221> MISC_FEATURE

<222> (259)..(259)

<223> Xaa is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp

<220>

<221> MISC_FEATURE

<222> (260)..(260)

<223> Xaa is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp

<400> 21

Met Asp Lys Ile Leu Gly Ala Ser Phe Leu Val Leu Trp Leu Gln Leu

1 5 10 15

Cys Trp Val Ser Gly Gln Gln Lys Glu Lys Ser Asp Gln Gln Gln Val

20 25 30

Lys Gln Ser Pro Gln Ser Leu Ile Val Gln Lys Gly Gly Ile Ser Ile

35 40 45

Ile Asn Cys Ala Tyr Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr

50 55 60

Gln Gln Phe Pro Gly Lys Gly Pro Ala Leu Leu Ile Ala Ile Arg Pro

65 70 75 80

Asp Val Ser Glu Lys Lys Glu Gly Arg Phe Thr Ile Ser Phe Asn Lys

85 90 95

Ser Ala Lys Gln Phe Ser Leu His Ile Met Asp Ser Gln Pro Gly Asp

100 105 110

Ser Ala Thr Tyr Phe Cys Ala Ala Glu Ala Gly Asn His Arg Gly Ser

115 120 125

Thr Leu Gly Arg Leu Tyr Phe Gly Arg Gly Thr Gln Leu Thr Val Trp

130 135 140

Pro Asn Ile Gln Asn Pro Glu Pro Ala Val Tyr Gln Leu Lys Asp Pro

145 150 155 160

Arg Ser Gln Asp Ser Thr Leu Cys Leu Phe Thr Asp Phe Asp Ser Gln

165 170 175

Ile Asn Val Pro Lys Thr Met Glu Ser Gly Thr Phe Ile Thr Asp Lys

180 185 190

Xaa Val Leu Asp Met Lys Ala Met Asp Ser Lys Ser Asn Gly Ala Ile

195 200 205

Ala Trp Ser Asn Gln Thr Ser Phe Thr Cys Gln Asp Ile Phe Lys Glu

210 215 220

Thr Asn Ala Thr Tyr Pro Ser Ser Asp Val Pro Cys Asp Ala Thr Leu

225 230 235 240

Thr Glu Lys Ser Phe Glu Thr Asp Met Asn Leu Asn Phe Gln Asn Leu

245 250 255

Xaa Val Xaa Xaa Leu Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn

260 265 270

Leu Leu Met Thr Leu Arg Leu Trp Ser Ser

275 280

<210> 22

<211> 307

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> MISC_FEATURE

<222> (191)..(191)

<223> Xaa is Ser or Cys

<400> 22

Met Ala Ser Arg Leu Leu Cys Trp Val Leu Leu Cys Leu Leu Gly Ala

1 5 10 15

Gly Pro Val Lys Ala Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys

20 25 30

Thr Arg Gly Gln Gln Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His

35 40 45

Arg Ser Val Ser Trp Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe

50 55 60

Leu Phe Glu Tyr Phe Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro

65 70 75 80

Gly Arg Phe Ser Gly Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn

85 90 95

Val Ser Thr Leu Glu Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser

100 105 110

Ser Leu Ala Ala Gly Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly

115 120 125

Thr Arg Leu Thr Val Leu Glu Asp Leu Arg Asn Val Thr Pro Pro Lys

130 135 140

Val Ser Leu Phe Glu Pro Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys

145 150 155 160

Ala Thr Leu Val Cys Leu Ala Arg Gly Phe Phe Pro Asp His Val Glu

165 170 175

Leu Ser Trp Trp Val Asn Gly Lys Glu Val His Ser Gly Val Xaa Thr

180 185 190

Asp Pro Gln Ala Tyr Lys Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser

195 200 205

Arg Leu Arg Val Ser Ala Thr Phe Trp His Asn Pro Arg Asn His Phe

210 215 220

Arg Cys Gln Val Gln Phe His Gly Leu Ser Glu Glu Asp Lys Trp Pro

225 230 235 240

Glu Gly Ser Pro Lys Pro Val Thr Gln Asn Ile Ser Ala Glu Ala Trp

245 250 255

Gly Arg Ala Asp Cys Gly Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val

260 265 270

Leu Ser Ala Thr Ile Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu

275 280 285

Tyr Ala Val Leu Val Ser Thr Leu Val Val Met Ala Met Val Lys Arg

290 295 300

Lys Asn Ser

305

<210> 23

<211> 282

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 23

Met Asp Lys Ile Leu Gly Ala Ser Phe Leu Val Leu Trp Leu Gln Leu

1 5 10 15

Cys Trp Val Ser Gly Gln Gln Lys Glu Lys Ser Asp Gln Gln Gln Val

20 25 30

Lys Gln Ser Pro Gln Ser Leu Ile Val Gln Lys Gly Gly Ile Ser Ile

35 40 45

Ile Asn Cys Ala Tyr Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr

50 55 60

Gln Gln Phe Pro Gly Lys Gly Pro Ala Leu Leu Ile Ala Ile Arg Pro

65 70 75 80

Asp Val Ser Glu Lys Lys Glu Gly Arg Phe Thr Ile Ser Phe Asn Lys

85 90 95

Ser Ala Lys Gln Phe Ser Leu His Ile Met Asp Ser Gln Pro Gly Asp

100 105 110

Ser Ala Thr Tyr Phe Cys Ala Ala Glu Ala Gly Asn His Arg Gly Ser

115 120 125

Thr Leu Gly Arg Leu Tyr Phe Gly Arg Gly Thr Gln Leu Thr Val Trp

130 135 140

Pro Asn Ile Gln Asn Pro Glu Pro Ala Val Tyr Gln Leu Lys Asp Pro

145 150 155 160

Arg Ser Gln Asp Ser Thr Leu Cys Leu Phe Thr Asp Phe Asp Ser Gln

165 170 175

Ile Asn Val Pro Lys Thr Met Glu Ser Gly Thr Phe Ile Thr Asp Lys

180 185 190

Thr Val Leu Asp Met Lys Ala Met Asp Ser Lys Ser Asn Gly Ala Ile

195 200 205

Ala Trp Ser Asn Gln Thr Ser Phe Thr Cys Gln Asp Ile Phe Lys Glu

210 215 220

Thr Asn Ala Thr Tyr Pro Ser Ser Asp Val Pro Cys Asp Ala Thr Leu

225 230 235 240

Thr Glu Lys Ser Phe Glu Thr Asp Met Asn Leu Asn Phe Gln Asn Leu

245 250 255

Ser Val Met Gly Leu Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn

260 265 270

Leu Leu Met Thr Leu Arg Leu Trp Ser Ser

275 280

<210> 24

<211> 307

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 24

Met Ala Ser Arg Leu Leu Cys Trp Val Leu Leu Cys Leu Leu Gly Ala

1 5 10 15

Gly Pro Val Lys Ala Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys

20 25 30

Thr Arg Gly Gln Gln Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His

35 40 45

Arg Ser Val Ser Trp Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe

50 55 60

Leu Phe Glu Tyr Phe Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro

65 70 75 80

Gly Arg Phe Ser Gly Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn

85 90 95

Val Ser Thr Leu Glu Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser

100 105 110

Ser Leu Ala Ala Gly Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly

115 120 125

Thr Arg Leu Thr Val Leu Glu Asp Leu Arg Asn Val Thr Pro Pro Lys

130 135 140

Val Ser Leu Phe Glu Pro Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys

145 150 155 160

Ala Thr Leu Val Cys Leu Ala Arg Gly Phe Phe Pro Asp His Val Glu

165 170 175

Leu Ser Trp Trp Val Asn Gly Lys Glu Val His Ser Gly Val Ser Thr

180 185 190

Asp Pro Gln Ala Tyr Lys Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser

195 200 205

Arg Leu Arg Val Ser Ala Thr Phe Trp His Asn Pro Arg Asn His Phe

210 215 220

Arg Cys Gln Val Gln Phe His Gly Leu Ser Glu Glu Asp Lys Trp Pro

225 230 235 240

Glu Gly Ser Pro Lys Pro Val Thr Gln Asn Ile Ser Ala Glu Ala Trp

245 250 255

Gly Arg Ala Asp Cys Gly Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val

260 265 270

Leu Ser Ala Thr Ile Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu

275 280 285

Tyr Ala Val Leu Val Ser Thr Leu Val Val Met Ala Met Val Lys Arg

290 295 300

Lys Asn Ser

305

<210> 25

<211> 27

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 25

Arg Ala Lys Arg Ser Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys

1 5 10 15

Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro

20 25

<210> 26

<211> 24

<212> PRT

<213> Homo sapiens

<400> 26

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Asp Gly Val Gly Lys

1 5 10 15

Ser Ala Leu Thr Ile Gln Leu Ile

20

<210> 27

<211> 24

<212> PRT

<213> Homo sapiens

<400> 27

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys

1 5 10 15

Ser Ala Leu Thr Ile Gln Leu Ile

20

<210> 28

<211> 9

<212> PRT

<213> Homo sapiens

<400> 28

Val Val Gly Ala Asp Gly Val Gly Lys

1 5

<210> 29

<211> 10

<212> PRT

<213> Homo sapiens

<400> 29

Val Val Val Gly Ala Asp Gly Val Gly Lys

1 5 10

<210> 30

<211> 9

<212> PRT

<213> Homo sapiens

<400> 30

Val Val Gly Ala Gly Gly Val Gly Lys

1 5

<210> 31

<211> 10

<212> PRT

<213> Homo sapiens

<400> 31

Val Val Val Gly Ala Gly Gly Val Gly Lys

1 5 10

<210> 32

<211> 117

<212> PRT

<213> Homo sapiens

<400> 32

Gln Gln Gln Val Lys Gln Ser Pro Gln Ser Leu Ile Val Gln Lys Gly

1 5 10 15

Gly Ile Ser Ile Ile Asn Cys Ala Tyr Glu Asn Thr Ala Phe Asp Tyr

20 25 30

Phe Pro Trp Tyr Gln Gln Phe Pro Gly Lys Gly Pro Ala Leu Leu Ile

35 40 45

Ala Ile Arg Pro Asp Val Ser Glu Lys Lys Glu Gly Arg Phe Thr Ile

50 55 60

Ser Phe Asn Lys Ser Ala Lys Gln Phe Ser Leu His Ile Met Asp Ser

65 70 75 80

Gln Pro Gly Asp Ser Ala Thr Tyr Phe Cys Ala Ala Glu Ala Gly Asn

85 90 95

His Arg Gly Ser Thr Leu Gly Arg Leu Tyr Phe Gly Arg Gly Thr Gln

100 105 110

Leu Thr Val Trp Pro

115

<210> 33

<211> 115

<212> PRT

<213> Homo sapiens

<400> 33

Lys Ala Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys Thr Arg Gly

1 5 10 15

Gln Gln Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His Arg Ser Val

20 25 30

Ser Trp Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe Leu Phe Glu

35 40 45

Tyr Phe Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro Gly Arg Phe

50 55 60

Ser Gly Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn Val Ser Thr

65 70 75 80

Leu Glu Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser Ser Leu Ala

85 90 95

Ala Gly Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu

100 105 110

Thr Val Leu

115

<210> 34

<211> 254

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> MISC_FEATURE

<222> (165)..(165)

<223> Xaa is Thr or Cys

<220>

<221> MISC_FEATURE

<222> (229)..(229)

<223> Xaa represents Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp

<220>

<221> MISC_FEATURE

<222> (231)..(231)

<223> Xaa is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp

<220>

<221> MISC_FEATURE

<222> (232)..(232)

<223> Xaa is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp

<400> 34

Gln Gln Gln Val Lys Gln Ser Pro Gln Ser Leu Ile Val Gln Lys Gly

1 5 10 15

Gly Ile Ser Ile Ile Asn Cys Ala Tyr Glu Asn Thr Ala Phe Asp Tyr

20 25 30

Phe Pro Trp Tyr Gln Gln Phe Pro Gly Lys Gly Pro Ala Leu Leu Ile

35 40 45

Ala Ile Arg Pro Asp Val Ser Glu Lys Lys Glu Gly Arg Phe Thr Ile

50 55 60

Ser Phe Asn Lys Ser Ala Lys Gln Phe Ser Leu His Ile Met Asp Ser

65 70 75 80

Gln Pro Gly Asp Ser Ala Thr Tyr Phe Cys Ala Ala Glu Ala Gly Asn

85 90 95

His Arg Gly Ser Thr Leu Gly Arg Leu Tyr Phe Gly Arg Gly Thr Gln

100 105 110

Leu Thr Val Trp Pro Asn Ile Gln Asn Pro Glu Pro Ala Val Tyr Gln

115 120 125

Leu Lys Asp Pro Arg Ser Gln Asp Ser Thr Leu Cys Leu Phe Thr Asp

130 135 140

Phe Asp Ser Gln Ile Asn Val Pro Lys Thr Met Glu Ser Gly Thr Phe

145 150 155 160

Ile Thr Asp Lys Xaa Val Leu Asp Met Lys Ala Met Asp Ser Lys Ser

165 170 175

Asn Gly Ala Ile Ala Trp Ser Asn Gln Thr Ser Phe Thr Cys Gln Asp

180 185 190

Ile Phe Lys Glu Thr Asn Ala Thr Tyr Pro Ser Ser Asp Val Pro Cys

195 200 205

Asp Ala Thr Leu Thr Glu Lys Ser Phe Glu Thr Asp Met Asn Leu Asn

210 215 220

Phe Gln Asn Leu Xaa Val Xaa Xaa Leu Arg Ile Leu Leu Leu Lys Val

225 230 235 240

Ala Gly Phe Asn Leu Leu Met Thr Leu Arg Leu Trp Ser Ser

245 250

<210> 35

<211> 288

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> MISC_FEATURE

<222> (172)..(172)

<223> Xaa is Ser or Cys

<400> 35

Lys Ala Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys Thr Arg Gly

1 5 10 15

Gln Gln Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His Arg Ser Val

20 25 30

Ser Trp Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe Leu Phe Glu

35 40 45

Tyr Phe Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro Gly Arg Phe

50 55 60

Ser Gly Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn Val Ser Thr

65 70 75 80

Leu Glu Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser Ser Leu Ala

85 90 95

Ala Gly Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu

100 105 110

Thr Val Leu Glu Asp Leu Arg Asn Val Thr Pro Pro Lys Val Ser Leu

115 120 125

Phe Glu Pro Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys Ala Thr Leu

130 135 140

Val Cys Leu Ala Arg Gly Phe Phe Pro Asp His Val Glu Leu Ser Trp

145 150 155 160

Trp Val Asn Gly Lys Glu Val His Ser Gly Val Xaa Thr Asp Pro Gln

165 170 175

Ala Tyr Lys Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser Arg Leu Arg

180 185 190

Val Ser Ala Thr Phe Trp His Asn Pro Arg Asn His Phe Arg Cys Gln

195 200 205

Val Gln Phe His Gly Leu Ser Glu Glu Asp Lys Trp Pro Glu Gly Ser

210 215 220

Pro Lys Pro Val Thr Gln Asn Ile Ser Ala Glu Ala Trp Gly Arg Ala

225 230 235 240

Asp Cys Gly Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val Leu Ser Ala

245 250 255

Thr Ile Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu Tyr Ala Val

260 265 270

Leu Val Ser Thr Leu Val Val Met Ala Met Val Lys Arg Lys Asn Ser

275 280 285

<210> 36

<211> 254

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 36

Gln Gln Gln Val Lys Gln Ser Pro Gln Ser Leu Ile Val Gln Lys Gly

1 5 10 15

Gly Ile Ser Ile Ile Asn Cys Ala Tyr Glu Asn Thr Ala Phe Asp Tyr

20 25 30

Phe Pro Trp Tyr Gln Gln Phe Pro Gly Lys Gly Pro Ala Leu Leu Ile

35 40 45

Ala Ile Arg Pro Asp Val Ser Glu Lys Lys Glu Gly Arg Phe Thr Ile

50 55 60

Ser Phe Asn Lys Ser Ala Lys Gln Phe Ser Leu His Ile Met Asp Ser

65 70 75 80

Gln Pro Gly Asp Ser Ala Thr Tyr Phe Cys Ala Ala Glu Ala Gly Asn

85 90 95

His Arg Gly Ser Thr Leu Gly Arg Leu Tyr Phe Gly Arg Gly Thr Gln

100 105 110

Leu Thr Val Trp Pro Asn Ile Gln Asn Pro Glu Pro Ala Val Tyr Gln

115 120 125

Leu Lys Asp Pro Arg Ser Gln Asp Ser Thr Leu Cys Leu Phe Thr Asp

130 135 140

Phe Asp Ser Gln Ile Asn Val Pro Lys Thr Met Glu Ser Gly Thr Phe

145 150 155 160

Ile Thr Asp Lys Thr Val Leu Asp Met Lys Ala Met Asp Ser Lys Ser

165 170 175

Asn Gly Ala Ile Ala Trp Ser Asn Gln Thr Ser Phe Thr Cys Gln Asp

180 185 190

Ile Phe Lys Glu Thr Asn Ala Thr Tyr Pro Ser Ser Asp Val Pro Cys

195 200 205

Asp Ala Thr Leu Thr Glu Lys Ser Phe Glu Thr Asp Met Asn Leu Asn

210 215 220

Phe Gln Asn Leu Ser Val Met Gly Leu Arg Ile Leu Leu Leu Lys Val

225 230 235 240

Ala Gly Phe Asn Leu Leu Met Thr Leu Arg Leu Trp Ser Ser

245 250

<210> 37

<211> 288

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 37

Lys Ala Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys Thr Arg Gly

1 5 10 15

Gln Gln Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His Arg Ser Val

20 25 30

Ser Trp Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe Leu Phe Glu

35 40 45

Tyr Phe Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro Gly Arg Phe

50 55 60

Ser Gly Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn Val Ser Thr

65 70 75 80

Leu Glu Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser Ser Leu Ala

85 90 95

Ala Gly Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu

100 105 110

Thr Val Leu Glu Asp Leu Arg Asn Val Thr Pro Pro Lys Val Ser Leu

115 120 125

Phe Glu Pro Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys Ala Thr Leu

130 135 140

Val Cys Leu Ala Arg Gly Phe Phe Pro Asp His Val Glu Leu Ser Trp

145 150 155 160

Trp Val Asn Gly Lys Glu Val His Ser Gly Val Ser Thr Asp Pro Gln

165 170 175

Ala Tyr Lys Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser Arg Leu Arg

180 185 190

Val Ser Ala Thr Phe Trp His Asn Pro Arg Asn His Phe Arg Cys Gln

195 200 205

Val Gln Phe His Gly Leu Ser Glu Glu Asp Lys Trp Pro Glu Gly Ser

210 215 220

Pro Lys Pro Val Thr Gln Asn Ile Ser Ala Glu Ala Trp Gly Arg Ala

225 230 235 240

Asp Cys Gly Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val Leu Ser Ala

245 250 255

Thr Ile Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu Tyr Ala Val

260 265 270

Leu Val Ser Thr Leu Val Val Met Ala Met Val Lys Arg Lys Asn Ser

275 280 285

<210> 38

<211> 137

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 38

Asn Ile Gln Asn Pro Glu Pro Ala Val Tyr Gln Leu Lys Asp Pro Arg

1 5 10 15

Ser Gln Asp Ser Thr Leu Cys Leu Phe Thr Asp Phe Asp Ser Gln Ile

20 25 30

Asn Val Pro Lys Thr Met Glu Ser Gly Thr Phe Ile Thr Asp Lys Cys

35 40 45

Val Leu Asp Met Lys Ala Met Asp Ser Lys Ser Asn Gly Ala Ile Ala

50 55 60

Trp Ser Asn Gln Thr Ser Phe Thr Cys Gln Asp Ile Phe Lys Glu Thr

65 70 75 80

Asn Ala Thr Tyr Pro Ser Ser Asp Val Pro Cys Asp Ala Thr Leu Thr

85 90 95

Glu Lys Ser Phe Glu Thr Asp Met Asn Leu Asn Phe Gln Asn Leu Leu

100 105 110

Val Ile Val Leu Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn Leu

115 120 125

Leu Met Thr Leu Arg Leu Trp Ser Ser

130 135

<210> 39

<211> 173

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 39

Glu Asp Leu Arg Asn Val Thr Pro Pro Lys Val Ser Leu Phe Glu Pro

1 5 10 15

Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys Ala Thr Leu Val Cys Leu

20 25 30

Ala Arg Gly Phe Phe Pro Asp His Val Glu Leu Ser Trp Trp Val Asn

35 40 45

Gly Lys Glu Val His Ser Gly Val Cys Thr Asp Pro Gln Ala Tyr Lys

50 55 60

Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser Arg Leu Arg Val Ser Ala

65 70 75 80

Thr Phe Trp His Asn Pro Arg Asn His Phe Arg Cys Gln Val Gln Phe

85 90 95

His Gly Leu Ser Glu Glu Asp Lys Trp Pro Glu Gly Ser Pro Lys Pro

100 105 110

Val Thr Gln Asn Ile Ser Ala Glu Ala Trp Gly Arg Ala Asp Cys Gly

115 120 125

Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val Leu Ser Ala Thr Ile Leu

130 135 140

Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu Tyr Ala Val Leu Val Ser

145 150 155 160

Thr Leu Val Val Met Ala Met Val Lys Arg Lys Asn Ser

165 170

<210> 40

<211> 282

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 40

Met Asp Lys Ile Leu Gly Ala Ser Phe Leu Val Leu Trp Leu Gln Leu

1 5 10 15

Cys Trp Val Ser Gly Gln Gln Lys Glu Lys Ser Asp Gln Gln Gln Val

20 25 30

Lys Gln Ser Pro Gln Ser Leu Ile Val Gln Lys Gly Gly Ile Ser Ile

35 40 45

Ile Asn Cys Ala Tyr Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr

50 55 60

Gln Gln Phe Pro Gly Lys Gly Pro Ala Leu Leu Ile Ala Ile Arg Pro

65 70 75 80

Asp Val Ser Glu Lys Lys Glu Gly Arg Phe Thr Ile Ser Phe Asn Lys

85 90 95

Ser Ala Lys Gln Phe Ser Leu His Ile Met Asp Ser Gln Pro Gly Asp

100 105 110

Ser Ala Thr Tyr Phe Cys Ala Ala Glu Ala Gly Asn His Arg Gly Ser

115 120 125

Thr Leu Gly Arg Leu Tyr Phe Gly Arg Gly Thr Gln Leu Thr Val Trp

130 135 140

Pro Asn Ile Gln Asn Pro Glu Pro Ala Val Tyr Gln Leu Lys Asp Pro

145 150 155 160

Arg Ser Gln Asp Ser Thr Leu Cys Leu Phe Thr Asp Phe Asp Ser Gln

165 170 175

Ile Asn Val Pro Lys Thr Met Glu Ser Gly Thr Phe Ile Thr Asp Lys

180 185 190

Cys Val Leu Asp Met Lys Ala Met Asp Ser Lys Ser Asn Gly Ala Ile

195 200 205

Ala Trp Ser Asn Gln Thr Ser Phe Thr Cys Gln Asp Ile Phe Lys Glu

210 215 220

Thr Asn Ala Thr Tyr Pro Ser Ser Asp Val Pro Cys Asp Ala Thr Leu

225 230 235 240

Thr Glu Lys Ser Phe Glu Thr Asp Met Asn Leu Asn Phe Gln Asn Leu

245 250 255

Leu Val Ile Val Leu Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn

260 265 270

Leu Leu Met Thr Leu Arg Leu Trp Ser Ser

275 280

<210> 41

<211> 307

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 41

Met Ala Ser Arg Leu Leu Cys Trp Val Leu Leu Cys Leu Leu Gly Ala

1 5 10 15

Gly Pro Val Lys Ala Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys

20 25 30

Thr Arg Gly Gln Gln Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His

35 40 45

Arg Ser Val Ser Trp Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe

50 55 60

Leu Phe Glu Tyr Phe Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro

65 70 75 80

Gly Arg Phe Ser Gly Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn

85 90 95

Val Ser Thr Leu Glu Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser

100 105 110

Ser Leu Ala Ala Gly Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly

115 120 125

Thr Arg Leu Thr Val Leu Glu Asp Leu Arg Asn Val Thr Pro Pro Lys

130 135 140

Val Ser Leu Phe Glu Pro Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys

145 150 155 160

Ala Thr Leu Val Cys Leu Ala Arg Gly Phe Phe Pro Asp His Val Glu

165 170 175

Leu Ser Trp Trp Val Asn Gly Lys Glu Val His Ser Gly Val Cys Thr

180 185 190

Asp Pro Gln Ala Tyr Lys Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser

195 200 205

Arg Leu Arg Val Ser Ala Thr Phe Trp His Asn Pro Arg Asn His Phe

210 215 220

Arg Cys Gln Val Gln Phe His Gly Leu Ser Glu Glu Asp Lys Trp Pro

225 230 235 240

Glu Gly Ser Pro Lys Pro Val Thr Gln Asn Ile Ser Ala Glu Ala Trp

245 250 255

Gly Arg Ala Asp Cys Gly Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val

260 265 270

Leu Ser Ala Thr Ile Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu

275 280 285

Tyr Ala Val Leu Val Ser Thr Leu Val Val Met Ala Met Val Lys Arg

290 295 300

Lys Asn Ser

305

<210> 42

<211> 254

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 42

Gln Gln Gln Val Lys Gln Ser Pro Gln Ser Leu Ile Val Gln Lys Gly

1 5 10 15

Gly Ile Ser Ile Ile Asn Cys Ala Tyr Glu Asn Thr Ala Phe Asp Tyr

20 25 30

Phe Pro Trp Tyr Gln Gln Phe Pro Gly Lys Gly Pro Ala Leu Leu Ile

35 40 45

Ala Ile Arg Pro Asp Val Ser Glu Lys Lys Glu Gly Arg Phe Thr Ile

50 55 60

Ser Phe Asn Lys Ser Ala Lys Gln Phe Ser Leu His Ile Met Asp Ser

65 70 75 80

Gln Pro Gly Asp Ser Ala Thr Tyr Phe Cys Ala Ala Glu Ala Gly Asn

85 90 95

His Arg Gly Ser Thr Leu Gly Arg Leu Tyr Phe Gly Arg Gly Thr Gln

100 105 110

Leu Thr Val Trp Pro Asn Ile Gln Asn Pro Glu Pro Ala Val Tyr Gln

115 120 125

Leu Lys Asp Pro Arg Ser Gln Asp Ser Thr Leu Cys Leu Phe Thr Asp

130 135 140

Phe Asp Ser Gln Ile Asn Val Pro Lys Thr Met Glu Ser Gly Thr Phe

145 150 155 160

Ile Thr Asp Lys Cys Val Leu Asp Met Lys Ala Met Asp Ser Lys Ser

165 170 175

Asn Gly Ala Ile Ala Trp Ser Asn Gln Thr Ser Phe Thr Cys Gln Asp

180 185 190

Ile Phe Lys Glu Thr Asn Ala Thr Tyr Pro Ser Ser Asp Val Pro Cys

195 200 205

Asp Ala Thr Leu Thr Glu Lys Ser Phe Glu Thr Asp Met Asn Leu Asn

210 215 220

Phe Gln Asn Leu Leu Val Ile Val Leu Arg Ile Leu Leu Leu Lys Val

225 230 235 240

Ala Gly Phe Asn Leu Leu Met Thr Leu Arg Leu Trp Ser Ser

245 250

<210> 43

<211> 288

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 43

Lys Ala Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys Thr Arg Gly

1 5 10 15

Gln Gln Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His Arg Ser Val

20 25 30

Ser Trp Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe Leu Phe Glu

35 40 45

Tyr Phe Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro Gly Arg Phe

50 55 60

Ser Gly Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn Val Ser Thr

65 70 75 80

Leu Glu Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser Ser Leu Ala

85 90 95

Ala Gly Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu

100 105 110

Thr Val Leu Glu Asp Leu Arg Asn Val Thr Pro Pro Lys Val Ser Leu

115 120 125

Phe Glu Pro Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys Ala Thr Leu

130 135 140

Val Cys Leu Ala Arg Gly Phe Phe Pro Asp His Val Glu Leu Ser Trp

145 150 155 160

Trp Val Asn Gly Lys Glu Val His Ser Gly Val Cys Thr Asp Pro Gln

165 170 175

Ala Tyr Lys Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser Arg Leu Arg

180 185 190

Val Ser Ala Thr Phe Trp His Asn Pro Arg Asn His Phe Arg Cys Gln

195 200 205

Val Gln Phe His Gly Leu Ser Glu Glu Asp Lys Trp Pro Glu Gly Ser

210 215 220

Pro Lys Pro Val Thr Gln Asn Ile Ser Ala Glu Ala Trp Gly Arg Ala

225 230 235 240

Asp Cys Gly Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val Leu Ser Ala

245 250 255

Thr Ile Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu Tyr Ala Val

260 265 270

Leu Val Ser Thr Leu Val Val Met Ala Met Val Lys Arg Lys Asn Ser

275 280 285

<210> 44

<211> 435

<212> DNA

<213> Homo sapiens

<400> 44

atggacaaga tcctgggcgc ctcttttctg gtgctgtggc tgcagctgtg ctgggtgtcc 60

ggacagcaga aggagaagtc tgatcagcag caggtgaagc agtctcccca gagcctgatc 120

gtgcagaagg gcggcatcag catcatcaac tgtgcctacg agaataccgc cttcgattac 180

tttccctggt atcagcagtt cccaggcaag ggacccgccc tgctgatcgc aatcaggcct 240

gacgtgagcg agaagaagga gggccgcttc acaatcagct ttaataagtc cgccaagcag 300

ttctccctgc acatcatgga cagccagccc ggcgattccg ccacctactt ttgtgcagca 360

gaggcaggaa accacagggg ctccacactg ggccggctgt atttcggcag aggcacccag 420

ctgacagtgt ggcct 435

<210> 45

<211> 402

<212> DNA

<213> Homo sapiens

<400> 45

atggcctcca ggctgctgtg ctgggtgctg ctgtgcctgc tgggagcagg accagtgaag 60

gcaggcgtga cccagacacc taggtacctg atcaagaccc gcggccagca ggtgacactg 120

tcttgcagcc caatcagcgg ccaccgctcc gtgtcttggt accagcagac cccaggacag 180

ggcctgcagt tcctgtttga gtatttctcc gagacacaga ggaacaaggg caatttccct 240

ggccggtttt ctggcagaca gtttagcaac tcccgctctg agatgaacgt gagcaccctg 300

gagctgggcg atagcgccct gtacctgtgc gccagctccc tggccgcagg aggctatttc 360

aacgagcagt tctttggacc aggaaccagg ctgacagtgc tg 402

<210> 46

<211> 1859

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic

<400> 46

ccatggcctc caggctgctg tgctgggtgc tgctgtgcct gctgggagca ggaccagtga 60

aggcaggcgt gacccagaca cctaggtacc tgatcaagac ccgcggccag caggtgacac 120

tgtcttgcag cccaatcagc ggccaccgct ccgtgtcttg gtaccagcag accccaggac 180

agggcctgca gttcctgttt gagtatttct ccgagacaca gaggaacaag ggcaatttcc 240

ctggccggtt ttctggcaga cagtttagca actcccgctc tgagatgaac gtgagcaccc 300

tggagctggg cgatagcgcc ctgtacctgt gcgccagctc cctggccgca ggaggctatt 360

tcaacgagca gttctttgga ccaggaacca ggctgacagt gctggaggac ctgagaaatg 420

tgaccccccc taaggtgtcc ctgtttgagc cttctaaggc cgagatcgcc aacaagcaga 480

aggccaccct ggtgtgcctg gcaaggggct tctttccaga tcacgtggag ctgagctggt 540

gggtgaatgg caaggaggtg cactccggcg tgtgcaccga cccacaggcc tacaaggaga 600

gcaactactc ctattgtctg tctagccggc tgagagtgtc cgccacattc tggcacaacc 660

caaggaatca cttccgctgc caggtgcagt ttcacggcct gagcgaggag gataagtggc 720

cagagggctc cccaaagcca gtgacccaga atatctctgc cgaggcatgg ggaagggcag 780

actgtggaat caccagcgcc tcctatcagc agggcgtgct gagcgccaca atcctgtacg 840

agatcctgct gggcaaggcc accctgtatg ccgtgctggt gtccacactg gtggtcatgg 900

ctatggtgaa gagaaagaac tctagggcaa agcggagcgg aagcggagca accaatttca 960

gcctgctgaa gcaggcaggc gatgtggagg agaaccctgg accaatggac aagatcctgg 1020

gcgcctcttt tctggtgctg tggctgcagc tgtgctgggt gtccggacag cagaaggaga 1080

agtctgatca gcagcaggtg aagcagtctc cccagagcct gatcgtgcag aagggcggca 1140

tcagcatcat caactgtgcc tacgagaata ccgccttcga ttactttccc tggtatcagc 1200

agttcccagg caagggaccc gccctgctga tcgcaatcag gcctgacgtg agcgagaaga 1260

aggagggccg cttcacaatc agctttaata agtccgccaa gcagttctcc ctgcacatca 1320

tggacagcca gcccggcgat tccgccacct acttttgtgc agcagaggca ggaaaccaca 1380

ggggctccac actgggccgg ctgtatttcg gcagaggcac ccagctgaca gtgtggccta 1440

acatccagaa tcccgagcct gccgtgtacc agctgaagga cccaagatcc caggattcta 1500

ccctgtgcct gttcacagac tttgattctc agatcaatgt gcctaagaca atggagagcg 1560

gcacctttat cacagacaag tgcgtgctgg acatgaaggc tatggactcc aagtctaacg 1620

gcgccatcgc ctggtctaat cagaccagct tcacatgcca ggatatcttt aaggagacaa 1680

acgccacata tccttcctct gacgtgccat gtgatgccac cctgacagag aagagcttcg 1740

agacagacat gaacctgaat tttcagaacc tgctggtcat cgtgctgcgg atcctgctgc 1800

tgaaggtggc cggcttcaat ctgctgatga cactgagact gtggagctcc tgagaattc 1859

<210> 47

<211> 616

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 47

Met Ala Ser Arg Leu Leu Cys Trp Val Leu Leu Cys Leu Leu Gly Ala

1 5 10 15

Gly Pro Val Lys Ala Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys

20 25 30

Thr Arg Gly Gln Gln Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His

35 40 45

Arg Ser Val Ser Trp Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe

50 55 60

Leu Phe Glu Tyr Phe Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro

65 70 75 80

Gly Arg Phe Ser Gly Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn

85 90 95

Val Ser Thr Leu Glu Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser

100 105 110

Ser Leu Ala Ala Gly Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly

115 120 125

Thr Arg Leu Thr Val Leu Glu Asp Leu Arg Asn Val Thr Pro Pro Lys

130 135 140

Val Ser Leu Phe Glu Pro Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys

145 150 155 160

Ala Thr Leu Val Cys Leu Ala Arg Gly Phe Phe Pro Asp His Val Glu

165 170 175

Leu Ser Trp Trp Val Asn Gly Lys Glu Val His Ser Gly Val Cys Thr

180 185 190

Asp Pro Gln Ala Tyr Lys Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser

195 200 205

Arg Leu Arg Val Ser Ala Thr Phe Trp His Asn Pro Arg Asn His Phe

210 215 220

Arg Cys Gln Val Gln Phe His Gly Leu Ser Glu Glu Asp Lys Trp Pro

225 230 235 240

Glu Gly Ser Pro Lys Pro Val Thr Gln Asn Ile Ser Ala Glu Ala Trp

245 250 255

Gly Arg Ala Asp Cys Gly Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val

260 265 270

Leu Ser Ala Thr Ile Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu

275 280 285

Tyr Ala Val Leu Val Ser Thr Leu Val Val Met Ala Met Val Lys Arg

290 295 300

Lys Asn Ser Arg Ala Lys Arg Ser Gly Ser Gly Ala Thr Asn Phe Ser

305 310 315 320

Leu Leu Lys Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Met Asp

325 330 335

Lys Ile Leu Gly Ala Ser Phe Leu Val Leu Trp Leu Gln Leu Cys Trp

340 345 350

Val Ser Gly Gln Gln Lys Glu Lys Ser Asp Gln Gln Gln Val Lys Gln

355 360 365

Ser Pro Gln Ser Leu Ile Val Gln Lys Gly Gly Ile Ser Ile Ile Asn

370 375 380

Cys Ala Tyr Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr Gln Gln

385 390 395 400

Phe Pro Gly Lys Gly Pro Ala Leu Leu Ile Ala Ile Arg Pro Asp Val

405 410 415

Ser Glu Lys Lys Glu Gly Arg Phe Thr Ile Ser Phe Asn Lys Ser Ala

420 425 430

Lys Gln Phe Ser Leu His Ile Met Asp Ser Gln Pro Gly Asp Ser Ala

435 440 445

Thr Tyr Phe Cys Ala Ala Glu Ala Gly Asn His Arg Gly Ser Thr Leu

450 455 460

Gly Arg Leu Tyr Phe Gly Arg Gly Thr Gln Leu Thr Val Trp Pro Asn

465 470 475 480

Ile Gln Asn Pro Glu Pro Ala Val Tyr Gln Leu Lys Asp Pro Arg Ser

485 490 495

Gln Asp Ser Thr Leu Cys Leu Phe Thr Asp Phe Asp Ser Gln Ile Asn

500 505 510

Val Pro Lys Thr Met Glu Ser Gly Thr Phe Ile Thr Asp Lys Cys Val

515 520 525

Leu Asp Met Lys Ala Met Asp Ser Lys Ser Asn Gly Ala Ile Ala Trp

530 535 540

Ser Asn Gln Thr Ser Phe Thr Cys Gln Asp Ile Phe Lys Glu Thr Asn

545 550 555 560

Ala Thr Tyr Pro Ser Ser Asp Val Pro Cys Asp Ala Thr Leu Thr Glu

565 570 575

Lys Ser Phe Glu Thr Asp Met Asn Leu Asn Phe Gln Asn Leu Leu Val

580 585 590

Ile Val Leu Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn Leu Leu

595 600 605

Met Thr Leu Arg Leu Trp Ser Ser

610 615

<210> 48

<211> 74

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 48

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Gly Val Gly Lys

1 5 10 15

Ser Ala Leu Thr Ile Gln Leu Ile Met Thr Glu Tyr Lys Leu Val Val

20 25 30

Val Gly Ala Gly Gly Val Gly Lys Ser Ala Leu Thr Ile Gln Leu Ile

35 40 45

Gln Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu

50 55 60

Tyr Ser Ala Met Arg Asp Gln Tyr Met Arg

65 70

<210> 49

<211> 295

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 49

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Asp Gly Val Gly Lys

1 5 10 15

Ser Ala Leu Thr Ile Gln Leu Ile Met Thr Glu Tyr Lys Leu Val Val

20 25 30

Val Gly Ala Val Gly Val Gly Lys Ser Ala Leu Thr Ile Gln Leu Ile

35 40 45

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Cys Gly Val Gly Lys

50 55 60

Ser Ala Leu Thr Ile Gln Leu Ile Met Thr Glu Tyr Lys Leu Val Val

65 70 75 80

Val Gly Ala Ala Gly Val Gly Lys Ser Ala Leu Thr Ile Gln Leu Ile

85 90 95

Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Ser Gly Val Gly Lys

100 105 110

Ser Ala Leu Thr Ile Gln Leu Ile Met Thr Glu Tyr Lys Leu Val Val

115 120 125

Val Gly Ala Gly Asp Val Gly Lys Ser Ala Leu Thr Ile Gln Leu Ile

130 135 140

Gln Met Thr Glu Tyr Lys Leu Val Val Val Gly Ala Gly Arg Val Gly

145 150 155 160

Lys Ser Ala Leu Thr Ile Gln Leu Ile Gln Met Thr Glu Tyr Lys Leu

165 170 175

Val Val Val Gly Ala Gly Val Val Gly Lys Ser Ala Leu Thr Ile Gln

180 185 190

Leu Ile Gln Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Arg

195 200 205

Glu Glu Tyr Ser Ala Met Arg Asp Gln Tyr Met Arg Glu Thr Cys Leu

210 215 220

Leu Asp Ile Leu Asp Thr Ala Gly Leu Glu Glu Tyr Ser Ala Met Arg

225 230 235 240

Asp Gln Tyr Met Arg Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala

245 250 255

Gly Lys Glu Glu Tyr Ser Ala Met Arg Asp Gln Tyr Met Arg Glu Thr

260 265 270

Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly His Glu Glu Tyr Ser Ala

275 280 285

Met Arg Asp Gln Tyr Met Arg

290 295

<210> 50

<211> 10

<212> PRT

<213> Homo sapiens

<400> 50

Lys Leu Val Val Val Gly Ala Asp Gly Val

1 5 10

<210> 51

<211> 145

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 51

Met Ala Lys Ile Leu Gly Ala Ser Phe Leu Val Leu Trp Leu Gln Leu

1 5 10 15

Cys Trp Val Ser Gly Gln Gln Lys Glu Lys Ser Asp Gln Gln Gln Val

20 25 30

Lys Gln Ser Pro Gln Ser Leu Ile Val Gln Lys Gly Gly Ile Ser Ile

35 40 45

Ile Asn Cys Ala Tyr Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr

50 55 60

Gln Gln Phe Pro Gly Lys Gly Pro Ala Leu Leu Ile Ala Ile Arg Pro

65 70 75 80

Asp Val Ser Glu Lys Lys Glu Gly Arg Phe Thr Ile Ser Phe Asn Lys

85 90 95

Ser Ala Lys Gln Phe Ser Leu His Ile Met Asp Ser Gln Pro Gly Asp

100 105 110

Ser Ala Thr Tyr Phe Cys Ala Ala Glu Ala Gly Asn His Arg Gly Ser

115 120 125

Thr Leu Gly Arg Leu Tyr Phe Gly Arg Gly Thr Gln Leu Thr Val Trp

130 135 140

Pro

145

<210> 52

<211> 134

<212> PRT

<213> Homo sapiens

<400> 52

Met Gly Ser Arg Leu Leu Cys Trp Val Leu Leu Cys Leu Leu Gly Ala

1 5 10 15

Gly Pro Val Lys Ala Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys

20 25 30

Thr Arg Gly Gln Gln Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His

35 40 45

Arg Ser Val Ser Trp Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe

50 55 60

Leu Phe Glu Tyr Phe Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro

65 70 75 80

Gly Arg Phe Ser Gly Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn

85 90 95

Val Ser Thr Leu Glu Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser

100 105 110

Ser Leu Ala Ala Gly Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly

115 120 125

Thr Arg Leu Thr Val Leu

130

<210> 53

<211> 282

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> MISC_FEATURE

<222> (193) .. (193)

<223> Xaa is Thr or Cys

<220>

<221> MISC_FEATURE

<222> (257)..(257)

<223> Xaa represents Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp

<220>

<221> MISC_FEATURE

<222> (259)..(259)

<223> Xaa is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp

<220>

<221> MISC_FEATURE

<222> (260)..(260)

<223> Xaa is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp

<400> 53

Met Ala Lys Ile Leu Gly Ala Ser Phe Leu Val Leu Trp Leu Gln Leu

1 5 10 15

Cys Trp Val Ser Gly Gln Gln Lys Glu Lys Ser Asp Gln Gln Gln Val

20 25 30

Lys Gln Ser Pro Gln Ser Leu Ile Val Gln Lys Gly Gly Ile Ser Ile

35 40 45

Ile Asn Cys Ala Tyr Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr

50 55 60

Gln Gln Phe Pro Gly Lys Gly Pro Ala Leu Leu Ile Ala Ile Arg Pro

65 70 75 80

Asp Val Ser Glu Lys Lys Glu Gly Arg Phe Thr Ile Ser Phe Asn Lys

85 90 95

Ser Ala Lys Gln Phe Ser Leu His Ile Met Asp Ser Gln Pro Gly Asp

100 105 110

Ser Ala Thr Tyr Phe Cys Ala Ala Glu Ala Gly Asn His Arg Gly Ser

115 120 125

Thr Leu Gly Arg Leu Tyr Phe Gly Arg Gly Thr Gln Leu Thr Val Trp

130 135 140

Pro Asn Ile Gln Asn Pro Glu Pro Ala Val Tyr Gln Leu Lys Asp Pro

145 150 155 160

Arg Ser Gln Asp Ser Thr Leu Cys Leu Phe Thr Asp Phe Asp Ser Gln

165 170 175

Ile Asn Val Pro Lys Thr Met Glu Ser Gly Thr Phe Ile Thr Asp Lys

180 185 190

Xaa Val Leu Asp Met Lys Ala Met Asp Ser Lys Ser Asn Gly Ala Ile

195 200 205

Ala Trp Ser Asn Gln Thr Ser Phe Thr Cys Gln Asp Ile Phe Lys Glu

210 215 220

Thr Asn Ala Thr Tyr Pro Ser Ser Asp Val Pro Cys Asp Ala Thr Leu

225 230 235 240

Thr Glu Lys Ser Phe Glu Thr Asp Met Asn Leu Asn Phe Gln Asn Leu

245 250 255

Xaa Val Xaa Xaa Leu Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn

260 265 270

Leu Leu Met Thr Leu Arg Leu Trp Ser Ser

275 280

<210> 54

<211> 307

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> MISC_FEATURE

<222> (191)..(191)

<223> Xaa is Ser or Cys

<400> 54

Met Gly Ser Arg Leu Leu Cys Trp Val Leu Leu Cys Leu Leu Gly Ala

1 5 10 15

Gly Pro Val Lys Ala Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys

20 25 30

Thr Arg Gly Gln Gln Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His

35 40 45

Arg Ser Val Ser Trp Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe

50 55 60

Leu Phe Glu Tyr Phe Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro

65 70 75 80

Gly Arg Phe Ser Gly Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn

85 90 95

Val Ser Thr Leu Glu Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser

100 105 110

Ser Leu Ala Ala Gly Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly

115 120 125

Thr Arg Leu Thr Val Leu Glu Asp Leu Arg Asn Val Thr Pro Pro Lys

130 135 140

Val Ser Leu Phe Glu Pro Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys

145 150 155 160

Ala Thr Leu Val Cys Leu Ala Arg Gly Phe Phe Pro Asp His Val Glu

165 170 175

Leu Ser Trp Trp Val Asn Gly Lys Glu Val His Ser Gly Val Xaa Thr

180 185 190

Asp Pro Gln Ala Tyr Lys Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser

195 200 205

Arg Leu Arg Val Ser Ala Thr Phe Trp His Asn Pro Arg Asn His Phe

210 215 220

Arg Cys Gln Val Gln Phe His Gly Leu Ser Glu Glu Asp Lys Trp Pro

225 230 235 240

Glu Gly Ser Pro Lys Pro Val Thr Gln Asn Ile Ser Ala Glu Ala Trp

245 250 255

Gly Arg Ala Asp Cys Gly Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val

260 265 270

Leu Ser Ala Thr Ile Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu

275 280 285

Tyr Ala Val Leu Val Ser Thr Leu Val Val Met Ala Met Val Lys Arg

290 295 300

Lys Asn Ser

305

<210> 55

<211> 282

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 55

Met Ala Lys Ile Leu Gly Ala Ser Phe Leu Val Leu Trp Leu Gln Leu

1 5 10 15

Cys Trp Val Ser Gly Gln Gln Lys Glu Lys Ser Asp Gln Gln Gln Val

20 25 30

Lys Gln Ser Pro Gln Ser Leu Ile Val Gln Lys Gly Gly Ile Ser Ile

35 40 45

Ile Asn Cys Ala Tyr Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr

50 55 60

Gln Gln Phe Pro Gly Lys Gly Pro Ala Leu Leu Ile Ala Ile Arg Pro

65 70 75 80

Asp Val Ser Glu Lys Lys Glu Gly Arg Phe Thr Ile Ser Phe Asn Lys

85 90 95

Ser Ala Lys Gln Phe Ser Leu His Ile Met Asp Ser Gln Pro Gly Asp

100 105 110

Ser Ala Thr Tyr Phe Cys Ala Ala Glu Ala Gly Asn His Arg Gly Ser

115 120 125

Thr Leu Gly Arg Leu Tyr Phe Gly Arg Gly Thr Gln Leu Thr Val Trp

130 135 140

Pro Asn Ile Gln Asn Pro Glu Pro Ala Val Tyr Gln Leu Lys Asp Pro

145 150 155 160

Arg Ser Gln Asp Ser Thr Leu Cys Leu Phe Thr Asp Phe Asp Ser Gln

165 170 175

Ile Asn Val Pro Lys Thr Met Glu Ser Gly Thr Phe Ile Thr Asp Lys

180 185 190

Thr Val Leu Asp Met Lys Ala Met Asp Ser Lys Ser Asn Gly Ala Ile

195 200 205

Ala Trp Ser Asn Gln Thr Ser Phe Thr Cys Gln Asp Ile Phe Lys Glu

210 215 220

Thr Asn Ala Thr Tyr Pro Ser Ser Asp Val Pro Cys Asp Ala Thr Leu

225 230 235 240

Thr Glu Lys Ser Phe Glu Thr Asp Met Asn Leu Asn Phe Gln Asn Leu

245 250 255

Ser Val Met Gly Leu Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn

260 265 270

Leu Leu Met Thr Leu Arg Leu Trp Ser Ser

275 280

<210> 56

<211> 307

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 56

Met Gly Ser Arg Leu Leu Cys Trp Val Leu Leu Cys Leu Leu Gly Ala

1 5 10 15

Gly Pro Val Lys Ala Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys

20 25 30

Thr Arg Gly Gln Gln Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His

35 40 45

Arg Ser Val Ser Trp Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe

50 55 60

Leu Phe Glu Tyr Phe Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro

65 70 75 80

Gly Arg Phe Ser Gly Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn

85 90 95

Val Ser Thr Leu Glu Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser

100 105 110

Ser Leu Ala Ala Gly Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly

115 120 125

Thr Arg Leu Thr Val Leu Glu Asp Leu Arg Asn Val Thr Pro Pro Lys

130 135 140

Val Ser Leu Phe Glu Pro Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys

145 150 155 160

Ala Thr Leu Val Cys Leu Ala Arg Gly Phe Phe Pro Asp His Val Glu

165 170 175

Leu Ser Trp Trp Val Asn Gly Lys Glu Val His Ser Gly Val Ser Thr

180 185 190

Asp Pro Gln Ala Tyr Lys Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser

195 200 205

Arg Leu Arg Val Ser Ala Thr Phe Trp His Asn Pro Arg Asn His Phe

210 215 220

Arg Cys Gln Val Gln Phe His Gly Leu Ser Glu Glu Asp Lys Trp Pro

225 230 235 240

Glu Gly Ser Pro Lys Pro Val Thr Gln Asn Ile Ser Ala Glu Ala Trp

245 250 255

Gly Arg Ala Asp Cys Gly Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val

260 265 270

Leu Ser Ala Thr Ile Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu

275 280 285

Tyr Ala Val Leu Val Ser Thr Leu Val Val Met Ala Met Val Lys Arg

290 295 300

Lys Asn Ser

305

<210> 57

<211> 282

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 57

Met Ala Lys Ile Leu Gly Ala Ser Phe Leu Val Leu Trp Leu Gln Leu

1 5 10 15

Cys Trp Val Ser Gly Gln Gln Lys Glu Lys Ser Asp Gln Gln Gln Val

20 25 30

Lys Gln Ser Pro Gln Ser Leu Ile Val Gln Lys Gly Gly Ile Ser Ile

35 40 45

Ile Asn Cys Ala Tyr Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr

50 55 60

Gln Gln Phe Pro Gly Lys Gly Pro Ala Leu Leu Ile Ala Ile Arg Pro

65 70 75 80

Asp Val Ser Glu Lys Lys Glu Gly Arg Phe Thr Ile Ser Phe Asn Lys

85 90 95

Ser Ala Lys Gln Phe Ser Leu His Ile Met Asp Ser Gln Pro Gly Asp

100 105 110

Ser Ala Thr Tyr Phe Cys Ala Ala Glu Ala Gly Asn His Arg Gly Ser

115 120 125

Thr Leu Gly Arg Leu Tyr Phe Gly Arg Gly Thr Gln Leu Thr Val Trp

130 135 140

Pro Asn Ile Gln Asn Pro Glu Pro Ala Val Tyr Gln Leu Lys Asp Pro

145 150 155 160

Arg Ser Gln Asp Ser Thr Leu Cys Leu Phe Thr Asp Phe Asp Ser Gln

165 170 175

Ile Asn Val Pro Lys Thr Met Glu Ser Gly Thr Phe Ile Thr Asp Lys

180 185 190

Cys Val Leu Asp Met Lys Ala Met Asp Ser Lys Ser Asn Gly Ala Ile

195 200 205

Ala Trp Ser Asn Gln Thr Ser Phe Thr Cys Gln Asp Ile Phe Lys Glu

210 215 220

Thr Asn Ala Thr Tyr Pro Ser Ser Asp Val Pro Cys Asp Ala Thr Leu

225 230 235 240

Thr Glu Lys Ser Phe Glu Thr Asp Met Asn Leu Asn Phe Gln Asn Leu

245 250 255

Leu Val Ile Val Leu Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn

260 265 270

Leu Leu Met Thr Leu Arg Leu Trp Ser Ser

275 280

<210> 58

<211> 307

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 58

Met Gly Ser Arg Leu Leu Cys Trp Val Leu Leu Cys Leu Leu Gly Ala

1 5 10 15

Gly Pro Val Lys Ala Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys

20 25 30

Thr Arg Gly Gln Gln Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His

35 40 45

Arg Ser Val Ser Trp Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe

50 55 60

Leu Phe Glu Tyr Phe Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro

65 70 75 80

Gly Arg Phe Ser Gly Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn

85 90 95

Val Ser Thr Leu Glu Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser

100 105 110

Ser Leu Ala Ala Gly Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly

115 120 125

Thr Arg Leu Thr Val Leu Glu Asp Leu Arg Asn Val Thr Pro Pro Lys

130 135 140

Val Ser Leu Phe Glu Pro Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys

145 150 155 160

Ala Thr Leu Val Cys Leu Ala Arg Gly Phe Phe Pro Asp His Val Glu

165 170 175

Leu Ser Trp Trp Val Asn Gly Lys Glu Val His Ser Gly Val Cys Thr

180 185 190

Asp Pro Gln Ala Tyr Lys Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser

195 200 205

Arg Leu Arg Val Ser Ala Thr Phe Trp His Asn Pro Arg Asn His Phe

210 215 220

Arg Cys Gln Val Gln Phe His Gly Leu Ser Glu Glu Asp Lys Trp Pro

225 230 235 240

Glu Gly Ser Pro Lys Pro Val Thr Gln Asn Ile Ser Ala Glu Ala Trp

245 250 255

Gly Arg Ala Asp Cys Gly Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val

260 265 270

Leu Ser Ala Thr Ile Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu

275 280 285

Tyr Ala Val Leu Val Ser Thr Leu Val Val Met Ala Met Val Lys Arg

290 295 300

Lys Asn Ser

305

<210> 59

<211> 124

<212> PRT

<213> Homo sapiens

<400> 59

Gln Gln Lys Glu Lys Ser Asp Gln Gln Gln Val Lys Gln Ser Pro Gln

1 5 10 15

Ser Leu Ile Val Gln Lys Gly Gly Ile Ser Ile Ile Asn Cys Ala Tyr

20 25 30

Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr Gln Gln Phe Pro Gly

35 40 45

Lys Gly Pro Ala Leu Leu Ile Ala Ile Arg Pro Asp Val Ser Glu Lys

50 55 60

Lys Glu Gly Arg Phe Thr Ile Ser Phe Asn Lys Ser Ala Lys Gln Phe

65 70 75 80

Ser Leu His Ile Met Asp Ser Gln Pro Gly Asp Ser Ala Thr Tyr Phe

85 90 95

Cys Ala Ala Glu Ala Gly Asn His Arg Gly Ser Thr Leu Gly Arg Leu

100 105 110

Tyr Phe Gly Arg Gly Thr Gln Leu Thr Val Trp Pro

115 120

<210> 60

<211> 113

<212> PRT

<213> Homo sapiens

<400> 60

Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys Thr Arg Gly Gln Gln

1 5 10 15

Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His Arg Ser Val Ser Trp

20 25 30

Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe Leu Phe Glu Tyr Phe

35 40 45

Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro Gly Arg Phe Ser Gly

50 55 60

Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn Val Ser Thr Leu Glu

65 70 75 80

Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser Ser Leu Ala Ala Gly

85 90 95

Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val

100 105 110

Leu

<210> 61

<211> 261

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> MISC_FEATURE

<222> (172)..(172)

<223> Xaa is Thr or Cys

<220>

<221> MISC_FEATURE

<222> (236)..(236)

<223> Xaa represents Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp

<220>

<221> MISC_FEATURE

<222> (238)..(238)

<223> Xaa is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp

<220>

<221> MISC_FEATURE

<222> (239)..(239)

<223> Xaa is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp

<400> 61

Gln Gln Lys Glu Lys Ser Asp Gln Gln Gln Val Lys Gln Ser Pro Gln

1 5 10 15

Ser Leu Ile Val Gln Lys Gly Gly Ile Ser Ile Ile Asn Cys Ala Tyr

20 25 30

Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr Gln Gln Phe Pro Gly

35 40 45

Lys Gly Pro Ala Leu Leu Ile Ala Ile Arg Pro Asp Val Ser Glu Lys

50 55 60

Lys Glu Gly Arg Phe Thr Ile Ser Phe Asn Lys Ser Ala Lys Gln Phe

65 70 75 80

Ser Leu His Ile Met Asp Ser Gln Pro Gly Asp Ser Ala Thr Tyr Phe

85 90 95

Cys Ala Ala Glu Ala Gly Asn His Arg Gly Ser Thr Leu Gly Arg Leu

100 105 110

Tyr Phe Gly Arg Gly Thr Gln Leu Thr Val Trp Pro Asn Ile Gln Asn

115 120 125

Pro Glu Pro Ala Val Tyr Gln Leu Lys Asp Pro Arg Ser Gln Asp Ser

130 135 140

Thr Leu Cys Leu Phe Thr Asp Phe Asp Ser Gln Ile Asn Val Pro Lys

145 150 155 160

Thr Met Glu Ser Gly Thr Phe Ile Thr Asp Lys Xaa Val Leu Asp Met

165 170 175

Lys Ala Met Asp Ser Lys Ser Asn Gly Ala Ile Ala Trp Ser Asn Gln

180 185 190

Thr Ser Phe Thr Cys Gln Asp Ile Phe Lys Glu Thr Asn Ala Thr Tyr

195 200 205

Pro Ser Ser Asp Val Pro Cys Asp Ala Thr Leu Thr Glu Lys Ser Phe

210 215 220

Glu Thr Asp Met Asn Leu Asn Phe Gln Asn Leu Xaa Val Xaa Xaa Leu

225 230 235 240

Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn Leu Leu Met Thr Leu

245 250 255

Arg Leu Trp Ser Ser

260

<210> 62

<211> 286

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<220>

<221> MISC_FEATURE

<222> (170)..(170)

<223> Xaa is Ser or Cys

<400> 62

Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys Thr Arg Gly Gln Gln

1 5 10 15

Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His Arg Ser Val Ser Trp

20 25 30

Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe Leu Phe Glu Tyr Phe

35 40 45

Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro Gly Arg Phe Ser Gly

50 55 60

Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn Val Ser Thr Leu Glu

65 70 75 80

Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser Ser Leu Ala Ala Gly

85 90 95

Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val

100 105 110

Leu Glu Asp Leu Arg Asn Val Thr Pro Pro Lys Val Ser Leu Phe Glu

115 120 125

Pro Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys Ala Thr Leu Val Cys

130 135 140

Leu Ala Arg Gly Phe Phe Pro Asp His Val Glu Leu Ser Trp Trp Val

145 150 155 160

Asn Gly Lys Glu Val His Ser Gly Val Xaa Thr Asp Pro Gln Ala Tyr

165 170 175

Lys Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser Arg Leu Arg Val Ser

180 185 190

Ala Thr Phe Trp His Asn Pro Arg Asn His Phe Arg Cys Gln Val Gln

195 200 205

Phe His Gly Leu Ser Glu Glu Asp Lys Trp Pro Glu Gly Ser Pro Lys

210 215 220

Pro Val Thr Gln Asn Ile Ser Ala Glu Ala Trp Gly Arg Ala Asp Cys

225 230 235 240

Gly Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val Leu Ser Ala Thr Ile

245 250 255

Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu Tyr Ala Val Leu Val

260 265 270

Ser Thr Leu Val Val Met Ala Met Val Lys Arg Lys Asn Ser

275 280 285

<210> 63

<211> 261

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 63

Gln Gln Lys Glu Lys Ser Asp Gln Gln Gln Val Lys Gln Ser Pro Gln

1 5 10 15

Ser Leu Ile Val Gln Lys Gly Gly Ile Ser Ile Ile Asn Cys Ala Tyr

20 25 30

Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr Gln Gln Phe Pro Gly

35 40 45

Lys Gly Pro Ala Leu Leu Ile Ala Ile Arg Pro Asp Val Ser Glu Lys

50 55 60

Lys Glu Gly Arg Phe Thr Ile Ser Phe Asn Lys Ser Ala Lys Gln Phe

65 70 75 80

Ser Leu His Ile Met Asp Ser Gln Pro Gly Asp Ser Ala Thr Tyr Phe

85 90 95

Cys Ala Ala Glu Ala Gly Asn His Arg Gly Ser Thr Leu Gly Arg Leu

100 105 110

Tyr Phe Gly Arg Gly Thr Gln Leu Thr Val Trp Pro Asn Ile Gln Asn

115 120 125

Pro Glu Pro Ala Val Tyr Gln Leu Lys Asp Pro Arg Ser Gln Asp Ser

130 135 140

Thr Leu Cys Leu Phe Thr Asp Phe Asp Ser Gln Ile Asn Val Pro Lys

145 150 155 160

Thr Met Glu Ser Gly Thr Phe Ile Thr Asp Lys Thr Val Leu Asp Met

165 170 175

Lys Ala Met Asp Ser Lys Ser Asn Gly Ala Ile Ala Trp Ser Asn Gln

180 185 190

Thr Ser Phe Thr Cys Gln Asp Ile Phe Lys Glu Thr Asn Ala Thr Tyr

195 200 205

Pro Ser Ser Asp Val Pro Cys Asp Ala Thr Leu Thr Glu Lys Ser Phe

210 215 220

Glu Thr Asp Met Asn Leu Asn Phe Gln Asn Leu Ser Val Met Gly Leu

225 230 235 240

Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn Leu Leu Met Thr Leu

245 250 255

Arg Leu Trp Ser Ser

260

<210> 64

<211> 286

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 64

Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys Thr Arg Gly Gln Gln

1 5 10 15

Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His Arg Ser Val Ser Trp

20 25 30

Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe Leu Phe Glu Tyr Phe

35 40 45

Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro Gly Arg Phe Ser Gly

50 55 60

Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn Val Ser Thr Leu Glu

65 70 75 80

Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser Ser Leu Ala Ala Gly

85 90 95

Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val

100 105 110

Leu Glu Asp Leu Arg Asn Val Thr Pro Pro Lys Val Ser Leu Phe Glu

115 120 125

Pro Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys Ala Thr Leu Val Cys

130 135 140

Leu Ala Arg Gly Phe Phe Pro Asp His Val Glu Leu Ser Trp Trp Val

145 150 155 160

Asn Gly Lys Glu Val His Ser Gly Val Ser Thr Asp Pro Gln Ala Tyr

165 170 175

Lys Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser Arg Leu Arg Val Ser

180 185 190

Ala Thr Phe Trp His Asn Pro Arg Asn His Phe Arg Cys Gln Val Gln

195 200 205

Phe His Gly Leu Ser Glu Glu Asp Lys Trp Pro Glu Gly Ser Pro Lys

210 215 220

Pro Val Thr Gln Asn Ile Ser Ala Glu Ala Trp Gly Arg Ala Asp Cys

225 230 235 240

Gly Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val Leu Ser Ala Thr Ile

245 250 255

Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu Tyr Ala Val Leu Val

260 265 270

Ser Thr Leu Val Val Met Ala Met Val Lys Arg Lys Asn Ser

275 280 285

<210> 65

<211> 261

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 65

Gln Gln Lys Glu Lys Ser Asp Gln Gln Gln Val Lys Gln Ser Pro Gln

1 5 10 15

Ser Leu Ile Val Gln Lys Gly Gly Ile Ser Ile Ile Asn Cys Ala Tyr

20 25 30

Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr Gln Gln Phe Pro Gly

35 40 45

Lys Gly Pro Ala Leu Leu Ile Ala Ile Arg Pro Asp Val Ser Glu Lys

50 55 60

Lys Glu Gly Arg Phe Thr Ile Ser Phe Asn Lys Ser Ala Lys Gln Phe

65 70 75 80

Ser Leu His Ile Met Asp Ser Gln Pro Gly Asp Ser Ala Thr Tyr Phe

85 90 95

Cys Ala Ala Glu Ala Gly Asn His Arg Gly Ser Thr Leu Gly Arg Leu

100 105 110

Tyr Phe Gly Arg Gly Thr Gln Leu Thr Val Trp Pro Asn Ile Gln Asn

115 120 125

Pro Glu Pro Ala Val Tyr Gln Leu Lys Asp Pro Arg Ser Gln Asp Ser

130 135 140

Thr Leu Cys Leu Phe Thr Asp Phe Asp Ser Gln Ile Asn Val Pro Lys

145 150 155 160

Thr Met Glu Ser Gly Thr Phe Ile Thr Asp Lys Cys Val Leu Asp Met

165 170 175

Lys Ala Met Asp Ser Lys Ser Asn Gly Ala Ile Ala Trp Ser Asn Gln

180 185 190

Thr Ser Phe Thr Cys Gln Asp Ile Phe Lys Glu Thr Asn Ala Thr Tyr

195 200 205

Pro Ser Ser Asp Val Pro Cys Asp Ala Thr Leu Thr Glu Lys Ser Phe

210 215 220

Glu Thr Asp Met Asn Leu Asn Phe Gln Asn Leu Leu Val Ile Val Leu

225 230 235 240

Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn Leu Leu Met Thr Leu

245 250 255

Arg Leu Trp Ser Ser

260

<210> 66

<211> 286

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 66

Gly Val Thr Gln Thr Pro Arg Tyr Leu Ile Lys Thr Arg Gly Gln Gln

1 5 10 15

Val Thr Leu Ser Cys Ser Pro Ile Ser Gly His Arg Ser Val Ser Trp

20 25 30

Tyr Gln Gln Thr Pro Gly Gln Gly Leu Gln Phe Leu Phe Glu Tyr Phe

35 40 45

Ser Glu Thr Gln Arg Asn Lys Gly Asn Phe Pro Gly Arg Phe Ser Gly

50 55 60

Arg Gln Phe Ser Asn Ser Arg Ser Glu Met Asn Val Ser Thr Leu Glu

65 70 75 80

Leu Gly Asp Ser Ala Leu Tyr Leu Cys Ala Ser Ser Leu Ala Ala Gly

85 90 95

Gly Tyr Phe Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val

100 105 110

Leu Glu Asp Leu Arg Asn Val Thr Pro Pro Lys Val Ser Leu Phe Glu

115 120 125

Pro Ser Lys Ala Glu Ile Ala Asn Lys Gln Lys Ala Thr Leu Val Cys

130 135 140

Leu Ala Arg Gly Phe Phe Pro Asp His Val Glu Leu Ser Trp Trp Val

145 150 155 160

Asn Gly Lys Glu Val His Ser Gly Val Cys Thr Asp Pro Gln Ala Tyr

165 170 175

Lys Glu Ser Asn Tyr Ser Tyr Cys Leu Ser Ser Arg Leu Arg Val Ser

180 185 190

Ala Thr Phe Trp His Asn Pro Arg Asn His Phe Arg Cys Gln Val Gln

195 200 205

Phe His Gly Leu Ser Glu Glu Asp Lys Trp Pro Glu Gly Ser Pro Lys

210 215 220

Pro Val Thr Gln Asn Ile Ser Ala Glu Ala Trp Gly Arg Ala Asp Cys

225 230 235 240

Gly Ile Thr Ser Ala Ser Tyr Gln Gln Gly Val Leu Ser Ala Thr Ile

245 250 255

Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu Tyr Ala Val Leu Val

260 265 270

Ser Thr Leu Val Val Met Ala Met Val Lys Arg Lys Asn Ser

275 280 285

<210> 67

<211> 616

<212> PRT

<213> Artificial sequence

<220>

<223> Synthetic

<400> 67

Met Ala Lys Ile Leu Gly Ala Ser Phe Leu Val Leu Trp Leu Gln Leu

1 5 10 15

Cys Trp Val Ser Gly Gln Gln Lys Glu Lys Ser Asp Gln Gln Gln Val

20 25 30

Lys Gln Ser Pro Gln Ser Leu Ile Val Gln Lys Gly Gly Ile Ser Ile

35 40 45

Ile Asn Cys Ala Tyr Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr

50 55 60

Gln Gln Phe Pro Gly Lys Gly Pro Ala Leu Leu Ile Ala Ile Arg Pro

65 70 75 80

Asp Val Ser Glu Lys Lys Glu Gly Arg Phe Thr Ile Ser Phe Asn Lys

85 90 95

Ser Ala Lys Gln Phe Ser Leu His Ile Met Asp Ser Gln Pro Gly Asp

100 105 110

Ser Ala Thr Tyr Phe Cys Ala Ala Glu Ala Gly Asn His Arg Gly Ser

115 120 125

Thr Leu Gly Arg Leu Tyr Phe Gly Arg Gly Thr Gln Leu Thr Val Trp

130 135 140

Pro Asn Ile Gln Asn Pro Glu Pro Ala Val Tyr Gln Leu Lys Asp Pro

145 150 155 160

Arg Ser Gln Asp Ser Thr Leu Cys Leu Phe Thr Asp Phe Asp Ser Gln

165 170 175

Ile Asn Val Pro Lys Thr Met Glu Ser Gly Thr Phe Ile Thr Asp Lys

180 185 190

Cys Val Leu Asp Met Lys Ala Met Asp Ser Lys Ser Asn Gly Ala Ile

195 200 205

Ala Trp Ser Asn Gln Thr Ser Phe Thr Cys Gln Asp Ile Phe Lys Glu

210 215 220

Thr Asn Ala Thr Tyr Pro Ser Ser Asp Val Pro Cys Asp Ala Thr Leu

225 230 235 240

Thr Glu Lys Ser Phe Glu Thr Asp Met Asn Leu Asn Phe Gln Asn Leu

245 250 255

Leu Val Ile Val Leu Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn

260 265 270

Leu Leu Met Thr Leu Arg Leu Trp Ser Ser Arg Ala Lys Arg Ser Gly

275 280 285

Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu

290 295 300

Glu Asn Pro Gly Pro Met Gly Ser Arg Leu Leu Cys Trp Val Leu Leu

305 310 315 320

Cys Leu Leu Gly Ala Gly Pro Val Lys Ala Gly Val Thr Gln Thr Pro

325 330 335

Arg Tyr Leu Ile Lys Thr Arg Gly Gln Gln Val Thr Leu Ser Cys Ser

340 345 350

Pro Ile Ser Gly His Arg Ser Val Ser Trp Tyr Gln Gln Thr Pro Gly

355 360 365

Gln Gly Leu Gln Phe Leu Phe Glu Tyr Phe Ser Glu Thr Gln Arg Asn

370 375 380

Lys Gly Asn Phe Pro Gly Arg Phe Ser Gly Arg Gln Phe Ser Asn Ser

385 390 395 400

Arg Ser Glu Met Asn Val Ser Thr Leu Glu Leu Gly Asp Ser Ala Leu

405 410 415

Tyr Leu Cys Ala Ser Ser Leu Ala Ala Gly Gly Tyr Phe Asn Glu Gln

420 425 430

Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu Glu Asp Leu Arg Asn

435 440 445

Val Thr Pro Pro Lys Val Ser Leu Phe Glu Pro Ser Lys Ala Glu Ile

450 455 460

Ala Asn Lys Gln Lys Ala Thr Leu Val Cys Leu Ala Arg Gly Phe Phe

465 470 475 480

Pro Asp His Val Glu Leu Ser Trp Trp Val Asn Gly Lys Glu Val His

485 490 495

Ser Gly Val Cys Thr Asp Pro Gln Ala Tyr Lys Glu Ser Asn Tyr Ser

500 505 510

Tyr Cys Leu Ser Ser Arg Leu Arg Val Ser Ala Thr Phe Trp His Asn

515 520 525

Pro Arg Asn His Phe Arg Cys Gln Val Gln Phe His Gly Leu Ser Glu

530 535 540

Glu Asp Lys Trp Pro Glu Gly Ser Pro Lys Pro Val Thr Gln Asn Ile

545 550 555 560

Ser Ala Glu Ala Trp Gly Arg Ala Asp Cys Gly Ile Thr Ser Ala Ser

565 570 575

Tyr Gln Gln Gly Val Leu Ser Ala Thr Ile Leu Tyr Glu Ile Leu Leu

580 585 590

Gly Lys Ala Thr Leu Tyr Ala Val Leu Val Ser Thr Leu Val Val Met

595 600 605

Ala Met Val Lys Arg Lys Asn Ser

610 615

<---

Claims (229)

1. A T cell receptor (TCR) comprising a complementarity determining region (CDR) 1 of an α chain comprising the amino acid sequence of SEQ ID NO: 1, a CDR2 of an α chain comprising the amino acid sequence of SEQ ID NO: 2, a CDR3 of an α chain comprising the amino acid sequence of SEQ ID NO: 3, a CDR1 of a β chain comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 of a β chain comprising the amino acid sequence of SEQ ID NO: 5, and a CDR3 of a β chain comprising the amino acid sequence of SEQ ID NO: 6, wherein said TCR has antigen specificity for the amino acid sequence of a mutant human RAS with a substitution of glycine at position 12 with aspartic acid, wherein the amino acid sequence of the mutant human RAS is the amino acid sequence of a mutant human Kirsten rat sarcoma viral oncogene homolog (KRAS), a mutant human Harvey rat sarcoma viral oncogene homolog (HRAS), or a mutant human neuroblastoma viral oncogene RAS homolog (NRAS), and where position 12 is determined by the wild-type human KRAS protein, wild-type human HRAS protein, or wild-type human NRAS protein, respectively. 2. The TCR of claim 1, wherein the amino acid sequence of the mutant human RAS is VVVGADGVGK (SEQ ID NO: 29) and where the HLA class I molecule is encoded by the HLA-A*11:01 allele. 3. TCR according to claim 1 or 2, comprising: (i) the amino acid sequence of SEQ ID NO: 7; (ii) the amino acid sequence of SEQ ID NO: 8; (iii) the amino acid sequence of SEQ ID NO: 51; (iv) the amino acid sequence of SEQ ID NO: 52; (v) the amino acid sequence of SEQ ID NO: 32; (vi) the amino acid sequence of SEQ ID NO: 33; (vii) the amino acid sequence of SEQ ID NO: 59; (viii) the amino acid sequence of SEQ ID NO: 60; or (ix) both (i) and (ii), both (i) and (iv), both (ii) and (iii), both (iii) and (iv), both (v) and (vi), both (v) and (viii), both (vi) and (vii), or both (vii) and (viii). 4. TCR according to any one of claims 1-3, comprising: (a) an α chain constant region comprising the amino acid sequence of SEQ ID NO: 17, in which: (i) X at position 48 of SEQ ID NO: 17 is Thr or Cys; (ii) X at position 112 of SEQ ID NO: 17 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 114 of SEQ ID NO: 17 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 115 of SEQ ID NO: 17 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (b) a β chain constant region comprising the amino acid sequence of SEQ ID NO: 18, wherein X at position 57 of SEQ ID NO: 18 is Ser or Cys; or (c) both (a) and (b). 5. A TCR according to any one of claims 1-4, comprising: (a) an α chain comprising the amino acid sequence of SEQ ID NO: 21, in which: (i) X at position 193 of SEQ ID NO: 21 is Thr or Cys; (ii) X at position 257 of SEQ ID NO: 21 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 259 of SEQ ID NO: 21 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 260 of SEQ ID NO: 21 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (b) a β chain comprising the amino acid sequence of SEQ ID NO: 22, wherein X at position 191 of SEQ ID NO: 22 is Ser or Cys; (c) an α chain comprising the amino acid sequence of SEQ ID NO: 53, in which: (i) X at position 193 of SEQ ID NO: 53 is Thr or Cys; (ii) X at position 257 of SEQ ID NO: 53 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 259 of SEQ ID NO: 53 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 260 of SEQ ID NO: 53 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (d) a β chain comprising the amino acid sequence of SEQ ID NO: 54, wherein X at position 191 of SEQ ID NO: 54 is Ser or Cys; (e) both (a) and (b), both (a) and (d), both (b) and (c), or both (c) and (d); (f) an α chain comprising the amino acid sequence of SEQ ID NO: 34, in which: (i) X at position 165 of SEQ ID NO: 34 is Thr or Cys; (ii) X at position 229 of SEQ ID NO: 34 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 231 of SEQ ID NO: 34 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 232 of SEQ ID NO: 34 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (g) a β chain comprising the amino acid sequence of SEQ ID NO: 35, wherein X at position 172 of SEQ ID NO: 35 is Ser or Cys; (h) an α chain comprising the amino acid sequence of SEQ ID NO: 61, in which: (i) X at position 172 of SEQ ID NO: 61 is Thr or Cys; (ii) X at position 236 of SEQ ID NO: 61 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 238 of SEQ ID NO: 61 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 239 of SEQ ID NO: 61 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (i) a β chain comprising the amino acid sequence of SEQ ID NO: 62, wherein X at position 170 of SEQ ID NO: 62 is Ser or Cys; (j) both (f) and (g) or both (h) and (i); (k) an α chain comprising the amino acid sequence of SEQ ID NO: 36; (l) a β chain comprising the amino acid sequence of SEQ ID NO: 37; (m) an α chain comprising the amino acid sequence of SEQ ID NO: 63; (n) a β chain comprising the amino acid sequence of SEQ ID NO: 64; (o) an α chain comprising the amino acid sequence of SEQ ID NO: 42; (p) a β chain comprising the amino acid sequence of SEQ ID NO: 43; (q) an α chain comprising the amino acid sequence of SEQ ID NO: 65; (r) a β chain comprising the amino acid sequence of SEQ ID NO: 66; (s) both (k) and (l), both (m) and (n), both (o) and (p), or both (q) and (r); (t) an α chain comprising the amino acid sequence of SEQ ID NO: 23; (u) a β chain comprising the amino acid sequence of SEQ ID NO: 24; (v) an α chain comprising the amino acid sequence of SEQ ID NO: 55; (w) a β chain comprising the amino acid sequence of SEQ ID NO: 56; (x) an α chain comprising the amino acid sequence of SEQ ID NO: 40; (y) a β chain comprising the amino acid sequence of SEQ ID NO: 41; (z) an α chain comprising the amino acid sequence of SEQ ID NO: 57; (aa) a β chain comprising the amino acid sequence of SEQ ID NO: 58; or (ab) both (t) and (u), both (t) and (w), both (u) and (v), both (v) and (w), both (x) and (y), both (x) and (aa), both (y) and (z) or both (z) and (aa). 6. A polypeptide for producing a recombinant TCR having antigen specificity for the amino acid sequence of a mutant human RAS with a substitution of glycine at position 12 with aspartic acid, wherein the amino acid sequence of the mutant human RAS is the amino acid sequence of a mutant human KRAS, a mutant human HRAS, or a mutant human NRAS, and wherein position 12 is determined by a wild-type human KRAS protein, a wild-type human HRAS protein, or a wild-type human NRAS protein, respectively, and wherein the polypeptide comprises an α chain CDR1 comprising the amino acid sequence of SEQ ID NO: 1, an α chain CDR2 comprising the amino acid sequence of SEQ ID NO: 2, an α chain CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a β chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a β chain CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a β chain CDR3 comprising the amino acid sequence of SEQ ID NO: 6. 7. The polypeptide according to claim 6, which comprises the amino acid sequence(s): (i) SEQ ID NO: 7; (ii) SEQ ID NO: 8; (iii) SEQ ID NO: 51; (iv) SEQ ID NO: 52; (v) SEQ ID NO: 32; (vi) SEQ ID NO: 33; (vii) SEQ ID NO: 59; (viii) SEQ ID NO: 60; or (ix) both (i) and (ii), both (i) and (iv), both (ii) and (iii), both (iii) and (iv), both (v) and (vi), both (v) and (viii), both (vi) and (vii) or both (vii) and (viii). 8. The polypeptide according to claim 6 or 7, comprising: (a) the amino acid sequence SEQ ID NO: 17, in which: (i) X at position 48 of SEQ ID NO: 17 is Thr or Cys; (ii) X at position 112 of SEQ ID NO: 17 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 114 of SEQ ID NO: 17 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 115 of SEQ ID NO: 17 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (b) the amino acid sequence of SEQ ID NO: 18, wherein X at position 57 of SEQ ID NO: 18 is Ser or Cys; or (c) both (a) and (b). 9. A polypeptide according to any one of paragraphs 6-8, comprising: (a) an α chain comprising the amino acid sequence of SEQ ID NO: 21, in which: (i) X at position 193 of SEQ ID NO: 21 is Thr or Cys; (ii) X at position 257 of SEQ ID NO: 21 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 259 of SEQ ID NO: 21 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 260 of SEQ ID NO: 21 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (b) a β chain comprising the amino acid sequence of SEQ ID NO: 22, wherein X at position 191 of SEQ ID NO: 22 is Ser or Cys; (c) an α chain comprising the amino acid sequence of SEQ ID NO: 53, in which: (i) X at position 193 of SEQ ID NO: 53 is Thr or Cys; (ii) X at position 257 of SEQ ID NO: 53 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 259 of SEQ ID NO: 53 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 260 of SEQ ID NO: 53 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (d) a β chain comprising the amino acid sequence of SEQ ID NO: 54, wherein X at position 191 of SEQ ID NO: 54 is Ser or Cys; (e) both (a) and (b), both (a) and (d), both (b) and (c), or both (c) and (d); (f) an α chain comprising the amino acid sequence of SEQ ID NO: 34, in which: (i) X at position 165 of SEQ ID NO: 34 is Thr or Cys; (ii) X at position 229 of SEQ ID NO: 34 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 231 of SEQ ID NO: 34 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 232 of SEQ ID NO: 34 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (g) a β chain comprising the amino acid sequence of SEQ ID NO: 35, wherein X at position 172 of SEQ ID NO: 35 is Ser or Cys; (h) an α chain comprising the amino acid sequence of SEQ ID NO: 61, in which: (i) X at position 172 of SEQ ID NO: 61 is Thr or Cys; (ii) X at position 236 of SEQ ID NO: 61 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 238 of SEQ ID NO: 61 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 239 of SEQ ID NO: 61 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (i) a β chain comprising the amino acid sequence of SEQ ID NO: 62, wherein X at position 170 of SEQ ID NO: 62 is Ser or Cys; (j) both (f) and (g) or both (h) and (i); (k) an α chain comprising the amino acid sequence of SEQ ID NO: 36; (I) a β chain comprising the amino acid sequence of SEQ ID NO: 37; (m) an α chain comprising the amino acid sequence of SEQ ID NO: 63; (n) a β chain comprising the amino acid sequence of SEQ ID NO: 64; (o) an α chain comprising the amino acid sequence of SEQ ID NO: 42; (p) a β chain comprising the amino acid sequence of SEQ ID NO: 43; (q) an α chain comprising the amino acid sequence of SEQ ID NO: 65; (r) a β chain comprising the amino acid sequence of SEQ ID NO: 66; (s) both (k) and (l), both (m) and (n), both (o) and (p), or both (q) and (r); (t) an α chain comprising the amino acid sequence of SEQ ID NO: 23; (u) a β chain comprising the amino acid sequence of SEQ ID NO: 24; (v) an α chain comprising the amino acid sequence of SEQ ID NO: 55; (w) a β chain comprising the amino acid sequence of SEQ ID NO: 56; (x) an α chain comprising the amino acid sequence of SEQ ID NO: 40; (y) a β chain comprising the amino acid sequence of SEQ ID NO: 41; (z) an α chain comprising the amino acid sequence of SEQ ID NO: 57; (aa) a β chain comprising the amino acid sequence of SEQ ID NO: 58; or (ab) both (t) and (u), both (t) and (w), both (u) and (v), both (v) and (w), both (x) and (y), both (x) and (aa), both (y) and (z) and both (z) and (aa). 10. A protein comprising a first polypeptide chain comprising a complementarity determining region (CDR) 1 of a TCR α chain comprising the amino acid sequence of SEQ ID NO: 1, a TCR α chain CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a TCR α chain CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and a second polypeptide chain comprising a TCR β chain CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a TCR β chain CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a TCR β chain CDR3 comprising the amino acid sequence of SEQ ID NO: 6, wherein the protein has antigen specificity for the amino acid sequence of a mutant human RAS with a substitution of glycine at position 12 with aspartic acid, wherein the amino acid sequence of the mutant human RAS is the amino acid sequence of a mutant human Kirsten rat sarcoma viral oncogene homolog (KRAS), a mutant human Harvey rat sarcoma viral oncogene homolog (HRAS), or a mutant human neuroblastoma viral oncogene RAS homolog (NRAS), and where position 12 is determined by the wild-type human KRAS protein, wild-type human HRAS protein, or wild-type human NRAS protein, respectively. 11. The protein according to claim 10, wherein: (i) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 7; (ii) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 8; (iii) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 51; (iv) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 52; (v) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 32; (vi) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 33; (vii) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 59; (viii) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 60; or (ix) both (i) and (ii), both (i) and (iv), both (ii) and (iii), both (iii) and (iv), both (v) and (vi), both (v) and (viii), both (vi) and (vii) or both (vii) and (viii). 12. The protein according to claim 10 or 11, wherein: (a) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 17, in which: (i) X at position 48 of SEQ ID NO: 17 is Thr or Cys; (ii) X at position 112 of SEQ ID NO: 17 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 114 of SEQ ID NO: 17 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 115 of SEQ ID NO: 17 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (b) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 18, wherein X at position 57 of SEQ ID NO: 18 is Ser or Cys; or (c) both (a) and (b). 13. The protein according to any one of paragraphs 10-12, in which: (a) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 21, in which: (i) X at position 193 of SEQ ID NO: 21 is Thr or Cys; (ii) X at position 257 of SEQ ID NO: 21 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 259 of SEQ ID NO: 21 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 260 of SEQ ID NO: 21 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (b) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 22, wherein X at position 191 of SEQ ID NO: 22 is Ser or Cys; (c) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 53, in which: (i) X at position 193 of SEQ ID NO: 53 is Thr or Cys; (ii) X at position 257 of SEQ ID NO: 53 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 259 of SEQ ID NO: 53 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 260 of SEQ ID NO: 53 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (d) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 45, wherein X at position 191 of SEQ ID NO: 54 is Ser or Cys; (e) both (a) and (b), both (a) and (d), both (b) and (c), or both (c) and (d); (f) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 34, in which: (i) X at position 165 of SEQ ID NO: 34 is Thr or Cys; (ii) X at position 229 of SEQ ID NO: 34 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 231 of SEQ ID NO: 34 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 232 of SEQ ID NO: 34 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (g) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 35, wherein X at position 172 of SEQ ID NO: 35 is Ser or Cys; (h) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, in which: (i) X at position 172 of SEQ ID NO: 61 is Thr or Cys; (ii) X at position 236 of SEQ ID NO: 61 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (iii) X at position 238 of SEQ ID NO: 61 is Met, Ala, Val, Leu, Ile, Pro, Phe or Trp; and (iv) X at position 239 of SEQ ID NO: 61 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met or Trp; (i) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 62, wherein X at position 170 of SEQ ID NO: 62 is Ser or Cys; (j) both (f) and (g) or both (h) and (i); (k) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 36; (I) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 37; (m) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 63; (n) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 64; (o) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 42; (p) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 43; (q) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 65; (r) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 66; (s) both (k) and (l), both (m) and (n), both (o) and (p), or both (q) and (r); (t) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 23; (u) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 24; (v) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 55; (w) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 56; (x) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 40; (y) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 41; (z) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 57; (aa) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 58; or (ab) both (t) and (u), both (t) and (w), both (u) and (v), both (v) and (w), both (x) and (y), both (x) and (aa), both (y) and (z) or both (z) and (aa). 14. A nucleic acid for expression by a host cell, wherein the nucleic acid comprises a nucleotide sequence encoding a TCR according to any one of claims 1-5. 15. A nucleic acid for expression by a host cell, wherein the nucleic acid comprises a nucleotide sequence encoding a polypeptide according to any one of claims 6 to 9. 16. A nucleic acid for expression by a host cell, wherein the nucleic acid comprises a nucleotide sequence encoding a protein according to any one of claims 10-13. 17. A recombinant expression vector containing the nucleic acid of claim 14. 18. A recombinant expression vector containing the nucleic acid of claim 15. 19. A recombinant expression vector containing the nucleic acid of claim 16. 20. A host cell for expressing on its surface a TCR having antigen specificity for the amino acid sequence of a mutant human RAS with a substitution of glycine at position 12 with aspartic acid, wherein the amino acid sequence of the mutant human RAS is the amino acid sequence of mutant human KRAS, mutant human HRAS, or mutant human NRAS, and wherein position 12 is determined by a wild-type human KRAS protein, a wild-type human HRAS protein, or a wild-type human NRAS protein, respectively, and wherein the host cell comprises the nucleic acid of claim 14 or the recombinant expression vector of claim 17. 21. A host cell for expressing on its surface a polypeptide for producing a recombinant TCR having antigen specificity for the amino acid sequence of a mutant human RAS with a substitution of glycine at position 12 with aspartic acid, wherein the amino acid sequence of the mutant human RAS is the amino acid sequence of mutant human KRAS, mutant human HRAS, or mutant human NRAS, and wherein position 12 is determined by a wild-type human KRAS protein, a wild-type human HRAS protein, or a wild-type human NRAS protein, respectively, and wherein the host cell comprises the nucleic acid of claim 15 or the recombinant expression vector of claim 18. 22. A host cell for expressing on its surface a protein having antigen specificity for the amino acid sequence of mutant human RAS with the substitution of glycine at position 12 with aspartic acid, wherein the amino acid sequence of the mutant human RAS is the amino acid sequence of mutant human KRAS, mutant human HRAS, or mutant human NRAS, and wherein position 12 is determined by a wild-type human KRAS protein, a wild-type human HRAS protein, or a wild-type human NRAS protein, respectively, and wherein the host cell comprises the nucleic acid of claim 16 or the recombinant expression vector of claim 19. 23. A population of cells for expression on their surface of a TCR with antigen specificity for the amino acid sequence of a mutant human RAS with a substitution of glycine at position 12 with aspartic acid, wherein the amino acid sequence of the mutant human RAS is the amino acid sequence of mutant human KRAS, mutant human HRAS, or mutant human NRAS, and wherein position 12 is determined by a wild-type human KRAS protein, a wild-type human HRAS protein, or a wild-type human NRAS protein, respectively, and wherein the population of cells comprises a host cell according to claim 20. 24. A population of cells for expression on their surface of a polypeptide for the production of a recombinant TCR having antigen specificity for the amino acid sequence of a mutant human RAS with the substitution of glycine at position 12 with aspartic acid, wherein the amino acid sequence of the mutant human RAS is the amino acid sequence of mutant human KRAS, mutant human HRAS, or mutant human NRAS, and wherein position 12 is determined by a wild-type human KRAS protein, a wild-type human HRAS protein, or a wild-type human NRAS protein, respectively, and wherein the population of cells comprises a host cell according to claim 21. 25. A population of cells for expression on their surface of a protein with antigen specificity for the amino acid sequence of mutant human RAS with a substitution of glycine at position 12 with aspartic acid, wherein the amino acid sequence of the mutant human RAS is the amino acid sequence of mutant human KRAS, mutant human HRAS, or mutant human NRAS, and wherein position 12 is determined by a wild-type human KRAS protein, a wild-type human HRAS protein, or a wild-type human NRAS protein, respectively, and wherein the population of cells comprises a host cell according to claim 22. 26. A pharmaceutical composition for treating cancer in a mammal, wherein the pharmaceutical composition comprises (a) a TCR according to any one of claims 1-5, a polypeptide according to any one of claims 6-9, a protein according to any one of claims 10-13, a nucleic acid according to any one of claims 14-16, a recombinant expression vector according to any one of claims 17-19, a host cell according to any one of claims 20-22 or a population of cells according to any one of claims 23-25 and (b) a pharmaceutically acceptable carrier, wherein the cancer expresses an amino acid sequence of mutant human RAS with a substitution of glycine at position 12 with aspartic acid, wherein the amino acid sequence of mutant human RAS is the amino acid sequence of mutant human KRAS, mutant human HRAS, or mutant human NRAS, and where position 12 is determined by the wild-type human KRAS protein, wild-type human HRAS protein, or wild-type human NRAS protein, respectively. 27. A method of treating cancer in a mammal, comprising administering to the mammal a TCR according to any one of claims 1-5, a polypeptide according to any one of claims 6-9, a protein according to any one of claims 10-13, a nucleic acid according to any one of claims 14-16, a recombinant expression vector according to any one of claims 17-19, a host cell according to any one of claims 20-22, a population of cells according to any one of claims 23-25 or a pharmaceutical composition according to claim 26, wherein the cancer expresses an amino acid sequence of a mutant human RAS with a substitution of glycine at position 12 with aspartic acid, wherein said amino acid sequence of mutant human RAS is the amino acid sequence of mutant human Kirsten rat sarcoma viral oncogene homolog (KRAS), mutant human Harvey rat sarcoma viral oncogene homolog (HRAS), or mutant human neuroblastoma viral oncogene RAS homolog (NRAS), and wherein position 12 is determined by wild-type human KRAS protein, wild-type human HRAS protein, or wild-type human NRAS protein, respectively. 28. A method for inducing an immune response to cancer in a mammal, comprising administering to the mammal a TCR according to any one of claims 1-5, a polypeptide according to any one of claims 6-9, a protein according to any one of claims 10-13, a nucleic acid according to any one of claims 14-16, a recombinant expression vector according to any one of claims 17-19, a host cell according to any one of claims 20-22, a population of cells according to any one of claims 23-25, or a pharmaceutical composition according to claim 26, wherein the cancer expresses an amino acid sequence of a mutant human RAS with a substitution of glycine at position 12 with aspartic acid, wherein said amino acid sequence of mutant human RAS is the amino acid sequence of mutant human Kirsten rat sarcoma viral oncogene homolog (KRAS), mutant human Harvey rat sarcoma viral oncogene homolog (HRAS), or mutant human neuroblastoma viral oncogene RAS homolog (NRAS), and where position 12 is determined by the wild-type human KRAS protein, wild-type human HRAS protein, or wild-type human NRAS protein, respectively. 29. The method according to claim 27 or 28, wherein the cancer is pancreatic cancer, colorectal cancer, lung cancer, endometrial cancer, ovarian cancer, or prostate cancer.