Anti-human ICOS antibodies for use in immunohistochemical (IHC) regiments and for

Abstract

In alternative embodiments, there is provided a chimeric, synthetic or recombinant anti-human ICOS protein or polypeptide (also referred to as inducible T-cell co-stimulatory factor or clustered differentiation-278 or CD278) antibody (Ab), including products of manufacture and kits comprising them; and methods for making and using them, including, for example, their use in the detection or diagnosis and treatment of cancer or other diseases or conditions involving ICOS expression. In alternative embodiments, an anti-ICOS protein (e.g., an antibody) as provided herein is used with an agent for determining whether ICOS expression or activity is present, increased, decreased or deleted. In alternative embodiments, anti-ICOS antibodies as provided herein are used in the diagnosis and / or treatment of cancer or tumors.

Description

[0001] Related Applications

[0002] This PCT International Utility Patent Application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Patent Application Serial No. (USSN) 63 / 409,076, filed on September 22, 2022. The entire contents of the above application are expressly incorporated herein by reference for all purposes. Technical Field

[0003] The present invention generally relates to immunohistochemistry (IHC) and cancer diagnosis and treatment. In alternative embodiments, chimeric, synthetic or recombinant anti-human ICOS protein or polypeptide (also referred to as inducible T cell co-stimulator or differentiation cluster-278 or CD278) antibodies (antibody, Ab) are provided, including manufacturing products and kits containing them; and methods for making and using them, including, for example, their use in the detection or diagnosis and treatment of cancer or other diseases or conditions involving ICOS expression. In alternative embodiments, anti-ICOS proteins (e.g., antibodies) as provided herein are used together with agents for determining whether ICOS expression or activity is present, increased, decreased or missing. In alternative embodiments, an anti-ICOS antibody as provided herein is used to diagnose and / or treat a cancer or tumor, e.g., a malignant tumor (optionally squamous cell carcinoma), breast cancer, colon cancer or colorectal cancer, melanoma (optionally malignant melanoma) or multiple myeloma, plasmacytoma, lymphoma, bladder cancer or urothelial carcinoma, cervical cancer, ovarian cancer, esophageal cancer or esophageal squamous cell carcinoma; malignant pleural mesothelioma, prostate cancer, microsatellite instability high / mismatch repair deficient tumors, human papillomavirus-positive or Epstein-Barr-positive tumors, hepatocellular carcinoma or liver cancer, lung cancer (optionally non-small cell lung cancer (NSCLC)), gastric cancer, renal / kidney cancer, pancreatic cancer, breast cancer (optionally triple-negative breast cancer), lymphoma or leukemia, or mycosis fungoides. Background Art

[0004] ICOS protein, also known as inducible T cell co-stimulator, cluster of differentiation-278 or CD278, belongs to the CD28 and CTLA-4 cell surface receptor family. ICOS protein forms homodimers and plays an important role in cell-cell signaling, immune response and cell proliferation regulation. ICOS is a co-stimulatory receptor for T cell enhancement.

[0005] ICOS+ T cells have been found in tumor tissues, and anti-CTLA-4 treatment of mice with ICOS+ tumors resulted in tumor rejection in 80% to 90% of subjects, but efficacy was less than 50% in genetically targeted mice lacking ICOS or its ligand (ICOSLG).

[0006] ICOS activation may enhance the effects of inhibitory checkpoint blockade, whereas its neutralization may reduce the function of immunosuppressive Tregs and inhibit lymphoid tumor cells expressing Tfh markers.

[0007] Vopratelimab (JTX-2011) is a humanized IgG1κ agonist mAb in development that specifically binds ICOS and is designed to enhance antitumor immune responses. The mechanism of action of vopareticlimab requires initial priming of T cells, followed by upregulation of ICOS expression on the cell surface of CD4 T cells, after which vopareticlimab engagement leads to T cell proliferation and sustained activation. Summary of the invention

[0008] In an alternative embodiment, an isolated or purified antibody (Ab) or antigen (Ag) binding fragment thereof, or a monomeric or dimeric antigen binding protein (ABP) is provided, which is capable of specifically binding to human ICOS (inducible T cell co-stimulatory) protein or human CD278, polypeptide,

[0009] The isolated or purified Ab or its Ag binding fragment, or monomer or dimer ABP comprises:

[0010] (a) a heavy chain variable region (VH), the VH comprising:

[0011] (1) an amino acid sequence comprising the three CDR1, CDR2 and CDR3 complementarity determining regions (CDRs) of SEQ ID NO: 1, or CDR1 amino acid (aa) residues GFSLSSYG (residues 25 to 32 of SEQ ID NO: 1), CDR2 aa residues INSDHST (residues 50 to 56 of SEQ ID NO: 1) and CDR3 aa residues ARSYGIGSIF (residues 93 to 102 of SEQ ID NO: 1), or

[0012] (2) an amino acid sequence having at least about 70%, 75%, 80%, 85%, 90%, 95%, 98% sequence identity, or between about 70% and 100% sequence identity to each of the three CDR1, CDR2, and CDR3 complementarity determining regions (CDRs) of SEQ ID NO: 1, or CDR1 amino acid (aa) residues GFSLSSYG (residues 25-32 of SEQ ID NO: 1), CDR2 aa residues INSDHST (residues 50 to 56 of SEQ ID NO: 1), and CDR3 aa residues ARSYGIGSIF (residues 93-102 of SEQ ID NO: 1), or

[0013] (3) an amino acid sequence having at least about 70%, 75%, 80%, 85%, 90%, 95%, 98% sequence identity, or between about 70% and 100% sequence identity to SEQ ID NO: 1, or an amino acid sequence having complete sequence identity to SEQ ID NO: 1; or

[0014] (b) a light chain variable region (VL), said VL comprising:

[0015] (1) an amino acid sequence comprising the three CDR1, CDR2 and CDR3 complementarity determining regions (CDRs) of SEQ ID NO: 2, or CDR1 amino acid (aa) residues KSVYNNNQ (residues 27 to 34 of SEQ ID NO: 2), CDR2 aa residues EAF (residues 52 to 54 of SEQ ID NO: 2), and CDR3 aa residues AAVYSDDSDNS (residues 91 to 101 of SEQ ID NO: 2), or

[0016] (2) an amino acid sequence having at least about 70%, 75%, 80%, 85%, 90%, 95%, 98% sequence identity, or between about 70% and 100% sequence identity to each of the three CDR1, CDR2, and CDR3 complementarity determining regions (CDRs) of SEQ ID NO:2, or CDR1 amino acid (aa) residues KSVYNNNQ (residues 27-34 of SEQ ID NO:2), CDR2 aa residues EAF (residues 52 to 54 of SEQ ID NO:2), and CDR3 aa residues AAVYSDDSDNS (residues 91-101 of SEQ ID NO:2), or

[0017] (3) an amino acid sequence having at least about 70%, 75%, 80%, 85%, 90%, 95%, 98% sequence identity, or between about 70% and 100% sequence identity to SEQ ID NO: 2, or an amino acid sequence having complete (100%) sequence identity to SEQ ID NO: 2; or

[0018] (c) The heavy chain variable region (VH) of (a) and the light chain variable region (VL) of (b).

[0019] In alternative embodiments of the isolated or purified antibody (Ab) or antigen (Ag) binding fragment thereof, or monomeric or dimeric antigen binding protein (ABP) as provided herein:

[0020] - The isolated or purified antibody (Ab) or antigen (Ag) binding fragment thereof, or monomeric or dimeric antigen binding protein (ABP) is manufactured or is in the following form:

[0021] Antigen binding fragment (Fab, or Ab fragment having only one constant domain and one variable domain of each of the Ab heavy and light chains),

[0022] F(ab')2 (or Ab digested by pepsin to produce the following two fragments: a F(ab')2 fragment and a pFc' (pepsin cleaved Fc) fragment),

[0023] Fab' (single chain of F(ab')2 fragment),

[0024] A single chain variable fragment (scFv) (or a fusion protein of the variable regions of the heavy and light chains of an Ab linked together with a linker peptide, optionally a linker peptide of about 10 to about 25 amino acids in length),

[0025] (scFv) 2, or two scFv or double scFv, or a single peptide chain having two variable heavy regions and two variable light regions to produce a tandem scFv,

[0026] Minibodies (or fusion proteins of the variable regions of the Ab heavy and light chains linked together with an alkyl, optionally methyl or ethyl group)

[0027] A diabody (or scFv, the scFv having a linker peptide that is too short for the two variable regions to fold together (optionally about 5 amino acids), thereby forcing the scFv to dimerize), a triabody or a tetrabody (or scFv, the scFv having a linker peptide that is too short for the two variable regions to fold together (optionally about 1 or 2 amino acids), thereby forcing the scFv to trimerize or tetramerize),

[0028] A single domain antibody (dAB) (or a single variable region of an Ab heavy chain or an Ab light chain), multiple complementarity determining region (CDR) fragments, or

[0029] Multispecific antibodies formed from two or more antibody fragments;

[0030] - The heavy chain variable region (VH), if present, comprises:

[0031] The following amino acid sequence:

[0032] QSLEESGGRLVTPGTPLTLTCTVS GFSLSSYG VSWVRQAPGKGLEWIGI INSDHST YYAKWAKGRFTISKTSTTVDLKITSPTTEDTATYFC ARSYGIGSIF WGPGTLVTVSS

[0033] (SEQ ID NO: 1), or

[0034] SEQ ID NO: 1 with one or more amino acid substitutions, additions (insertions) or deletions, and the recombinant Ab or its Ag-binding fragment or monomeric or dimeric ABP retains its ability to specifically bind to human ICOS protein or polypeptide;

[0035] - the one or more amino acid substitutions in the heavy chain variable region (VH) include one or more conservative amino acid substitutions, and optionally, the one or more conservative amino acid substitutions include: replacing an aliphatic amino acid with another aliphatic amino acid; replacing a serine with a threonine or vice versa; replacing an acidic residue with another acidic residue; replacing a residue with an amide group with another residue with an amide group; exchanging a basic residue with another basic residue; or replacing an aromatic residue with another aromatic residue;

[0036] - the heavy chain variable region further comprises at least a portion of a heavy chain constant region, and optionally, the heavy chain constant region comprises the following amino acid sequence:

[0037] GQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVTVTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVTSSSQPVTCNVAHPATNTKVDKTVAPSTCSKPTCPPPELLGGPSVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTA RPPLREQQFNSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPAPIEKTISKARGQPLEPKVYTMGPPREELSSRSVSLTCMINGFYPSDISVEWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSVPTSEWQRGDVFTCSVMHEALHNHYTQKSISRSPGK(SEQ ID NO:3);

[0038] - the heavy chain comprises a heavy chain variable region and a heavy chain constant region, and the heavy chain variable region and the heavy chain constant region comprise the following sequence:

[0039] QSLEESGGRLVTPGTPLTLTCTVSGFSLSSYGVSWVRQAPGKGLEWIGIINSDHSTYYAKGRFTISKTSTTVDLKITSPTTEDTATYFCARSYGIGSIFWGPGTLVT VSSGQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVTVTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSVTSSSQPVTCNVAHPATNTKVDKTVAPSTCSKPTCPP PELLGGPSVFIFPPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPAPIEKTISKARG QPLEPKVYTMGPPREELSSRSVSLTCMINGFYPSDISVEWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSVPTSEWQRGDVFTCSVMHEALHNHYTQKSISRSPGK(SEQ ID NO:4);

[0040] - the heavy chain variable region comprises an amino acid terminal signal sequence, and optionally, the heavy chain variable region amino acid terminal signal sequence comprises the sequence METGLRWLLLVAVLKGVQC (SEQ ID NO: 5);

[0041] -The heavy chain variable region with a signal sequence comprises the following sequence

[0042] METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLSSYGVSWVRQAPGKGLEWIGIINSDHSTYYAKWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARSYGIGSIFWGPGTLVTVSS(SEQ ID NO:6), or

[0043] The heavy chain, comprising the variable and constant domains and having a signal sequence, comprises the following sequence:

[0044] METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLSSYGVSWVRQAPGKGLEWIGIINSDHSTYYAKWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARS YGIGSIFWGPGTLVTVSSGQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVTVTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSVTSSSQPVTCNVAHPATNTKVDKTVAP STCSKPTCPPPELLGGPSVFIFPPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPAPIEKTI SKARGQPLEPKVYTMGPPREELSSRSVSLTCMINGFYPSDISVEWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSVPTSEWQRGDVFTCSVMHEALHNHYTQKSISRSPGK(SEQ ID NO:7);

[0045] - The light chain variable region (VL), if present, comprises:

[0046] (a) the following amino acid sequence:

[0047] AAVLTQTPSPVSAAVGGTVSISCQSS KSVYNNNQ LSWFQQKPGQRPKLLIY EAF KLPSGVPSRFKGSGSGTQFTLTISDVQCDDAATYYC AAVYSDDSDNSFGGGTEVVVK (SEQ ID NO: 2), or

[0048] SEQ ID NO: 2 having one or more amino acid substitutions, additions (insertions) or deletions, and the recombinant Ab or its Ag-binding fragment or monomeric or dimeric ABP retains its ability to specifically bind to human ICOS protein or polypeptide;

[0049] - the one or more amino acid substitutions of the light chain variable region (VL) include one or more conservative amino acid substitutions, and optionally, the one or more conservative amino acid substitutions include: conservative substitutions include: replacing an aliphatic amino acid with another aliphatic amino acid; replacing a serine with a threonine or vice versa; replacing an acidic residue with another acidic residue; replacing a residue with an amide group with another residue with an amide group; exchanging a basic residue with another basic residue; or replacing an aromatic residue with another aromatic residue;

[0050] - the light chain variable region further comprises at least a portion of a light chain constant region, and optionally, the light chain constant region comprises the following amino acid sequence:

[0051] GDPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTVTWEVDGTT QTTGIENSKTPQNSADCTYNLSSTLTLTSTQYNSHKEYTCKVTQGTTSVV QSFNRGDC(SEQ ID NO:8);

[0052] - the light chain comprises a variable region and a constant region, wherein the variable region and the constant region comprise the following sequence:

[0053] AAVLTQTPSPVSAAVGGTVSISCQSSKSVYNNNQLSWFQQKPGQRPKLLIYEAFKLPSGVPSRFKGSGSGTQFTLTISDVQCDDAATYYCAAVYSDDSDNSFGGGTEVVV KGDPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTVTWEVDGTTQTTGIENSKTPQNSADCTYNLSSTLTLTSTQYNSHKEYTCKVTQGTTSVVQSFNRGDC(SEQID NO:9);

[0054] - the light chain variable domain further comprises an amino-terminal signal sequence, and optionally, the light chain variable domain amino-terminal signal sequence comprises the sequence MDTRAPTQLLGLLLLWLPGATF (SEQ ID NO: 10);

[0055] -The light chain variable domain with a signal sequence comprises

[0056] or

[0057] -Light chain with signal sequence comprising

[0058] MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVSISCQSSKSVYNNNQLSWFQQKPGQRPKLLIYEAFKLPSGVPSRFKGSGSGTQFTLTISDVQCDDAATYYCAAVYSDDS DNSFGGGTEVVVKGDPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTVTWEVDGTTQTTGIENSKTPQNSADCTYNLSSTLTLTSTQYNSHKEYTCKVTQGTTSVVQSFNRGDC(SEQ ID NO:12);

[0059] - wherein SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 11 and / or SEQ ID NO: 12 have two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen or fifteen conservative amino acid substitutions, and the recombinant Ab or its Ag binding fragment or monomer or dimeric ABP retains its ability to specifically bind to human ICOS protein or polypeptide;

[0060] - the heavy chain constant region comprises an amino acid sequence from an IgG, IgM, IgA, IgD or IgE isotype; or the light chain constant region comprises an amino acid sequence from a kappa (κ) or lambda (λ) isotype;

[0061] - at least a portion of the heavy chain constant region, at least a portion of the light chain constant region, or at least a portion of the heavy chain constant region and the light chain constant region is or comprises an amino acid sequence of human, rabbit, mouse or rat origin, or comprises a constant region amino acid sequence derived from human, rabbit, mouse or rat;

[0062] - wherein said at least a portion of said heavy chain constant region, said at least a portion of said light chain constant region, or said at least a portion of said heavy chain constant region and said light chain constant region is or comprises a synthetic amino acid sequence;

[0063] - wherein the recombinant Ab, its Ab binding fragment, or monomeric or dimeric ABP, or the heavy chain constant region, or the light chain constant region, or the heavy chain constant region and the light chain constant region further comprise or are bound to a heterologous protein, peptide, or compound or composition, and optionally the heterologous protein or peptide, or compound or composition comprises a detectable protein, a detectable agent or a binding moiety; and optionally the heterologous protein or peptide comprises a carrier protein, or the heterologous protein, peptide or the compound or composition is covalently conjugated to the recombinant antibody (Ab) or its Ag binding fragment, or monomeric or dimeric ABP;

[0064] - the detectable agent or binding moiety comprises biotin, a fluorescent or chemiluminescent label, a fluorophore, perylene, fluorenyl, coumarin, 7-methoxycoumarin (Mca), 4-(dimethylaminoazo)benzene-4-carboxylic acid (dabcyl), Tamra, boron dipyrromethene (BODIPY) or a derivative thereof, a dye, a radioisotope, quantum dots or photoluminescent aqueous nanocrystals, a hapten or an antibody binding epitope or domain, and optionally, the dye is or comprises rhodamine, [2-(4 -nitro-2,1,3-benzoxadiazol-7-yl)aminoethyl]trimethylammonium (NBD), Nile Red or Nile Blue, or a fluorescent dye including sulfoindocyanine, and optionally the fluorophore is or comprises a dansyl, fluorescein, carboxyfluorescein (FAM) or 6-FAM moiety, and optionally the dye is or comprises a cyanine dye, Cy3 or Cy5, and optionally the hapten is or comprises a biotin, theophylline, digitoxin, carborane, fluorescein or bromodeoxyuridine moiety; and / or

[0065] - The Ab or its Ag-binding fragment, or monomer or dimeric ABP is a recombinant Ab or its Ag-binding fragment, or monomer or dimeric ABP, or comprises a peptide or polypeptide produced by recombinant technology.

[0066] In alternative embodiments, a chimeric or recombinant nucleic acid is provided that comprises or consists of a nucleic acid sequence encoding an Ab or Ag-binding fragment thereof, or a monomeric or dimeric ABP as provided herein.

[0067] In alternative embodiments of the chimeric or recombinant nucleic acids as provided herein:

[0068] - the chimeric or recombinant nucleic acid further comprises and is operably linked to a transcriptional regulatory element, and optionally, the transcriptional regulatory element comprises a promoter, and optionally, the promoter is an inducible promoter or a constitutive promoter; and / or

[0069] - the chimeric or recombinant nucleic acid further comprises a sequence encoding an amino-terminal signal peptide, and optionally, the amino-terminal signal peptide comprises the following amino acid sequence: METGLRWLLLVAVLKGVQC (SEQ ID NO: 5); or MDTRAPTQLLGLLLLWLPGATF (SEQ ID NO: 10).

[0070] In alternative embodiments, an expression cassette, vector, recombinant virus, artificial chromosome, cosmid or plasmid comprising a chimeric or recombinant nucleic acid as provided herein is provided, which comprises a nucleic acid encoding an antibody (Ab) or antigen (Ag) binding fragment thereof or a monomeric or dimeric antigen binding protein (ABP) as provided herein.

[0071] In alternative embodiments, cells are provided, comprising or containing a chimeric or recombinant antibody or dimeric antigen binding protein as provided herein, a chimeric or recombinant nucleic acid as provided herein, or an expression cassette, vector, recombinant virus, artificial chromosome, cosmid or plasmid as provided herein. In alternative embodiments of cells as provided herein: the cell is a bacterial, fungal, mammalian, yeast, insect or plant cell; or, the mammalian cell is a human cell.

[0072] In an alternative embodiment, a method is provided for detecting the presence of human ICOS (inducible T-cell co-stimulatory) protein or human CD278 protein or polypeptide in or on a cell, tissue, organ or a portion of any of the foregoing, the method comprising:

[0073] (a) contacting the cell, tissue or organ, or a portion of any of the foregoing, with at least one Ab or Ag-binding fragment thereof, or monomeric or dimeric ABP as provided herein, and

[0074] (b) detecting specific binding of said at least one Ab or Ag-binding fragment thereof, or monomeric or dimeric ABP to a BCMA polypeptide in or on said cell, tissue or organ, or a portion of any of the foregoing,

[0075] Thereby detecting the presence of the human BCMA protein in or on the cell, tissue or organ or a part of any of the foregoing.

[0076] In an alternative embodiment of the method for detecting the presence of human ICOS as provided herein:

[0077] - said at least one Ab or Ag-binding fragment thereof or monomer or dimeric ABP comprises a variable heavy chain (VH) having an amino acid sequence comprising SEQ ID NO: 1 and a variable light chain (VL) having an amino acid sequence comprising SEQ ID NO: 2;

[0078] - said at least one Ab or Ag-binding fragment thereof or monomeric or dimeric ABP comprises a heavy chain having an amino acid sequence comprising SEQ ID NO: 4 and a light chain having an amino acid sequence comprising SEQ ID NO: 9;

[0079] - the method comprises (or further comprises) contacting the cell, tissue or organ, or a portion of any of the foregoing, with two Abs or Ag-binding fragments thereof or monomeric or dimeric ABPs, or with a mixture of two Abs or Ag-binding fragments thereof or monomeric or dimeric ABPs, and optionally, the contacting comprises using an immunohistochemistry (IHC) assay;

[0080] - the method further comprises contacting the Ab or its Ag-binding fragment or monomer or dimeric ABP that specifically binds to human ICOS protein or human CD278 protein or polypeptide with a detectable agent to indicate or signal the specific binding of the Ab or its Ag-binding fragment or monomer or dimeric ABP to human ICOS protein or human CD278 protein or polypeptide, and optionally the detectable agent specifically binds to the Ab or its Ag-binding fragment or monomer or dimeric ABP;

[0081] - the cell, tissue, organ or a portion of any of the foregoing is or comprises: an early B cell, a progenitor B cell, a pre-B lymphocyte, a mature B lymphocyte, a follicle center cell, or a cell in a tonsil, an organ, a germinal center of a lymph node, a bone marrow stem cell, a bone marrow hematopoietic cell, a lymphocyte, a parafollicular T lymphocyte, a subpopulation of parafollicular T lymphocytes, a hepatobiliary tubule cell, a glomerular cell, a proximal tubule cell, a mammary myoepithelial cell, a stromal cell surrounding or associated with an infiltrating tumor cell, a kidney cell, a cerebellar cell, a prostate cell, a pancreatic cell, a bone marrow cell or an epithelial cell, and optionally the epithelial cell is an epithelial cell of the brain, lung, intestine, kidney, breast or placenta, and optionally the organ is the liver, prostate, brain, pancreas, bladder, colon, esophagus, kidney, skin or lung, and optionally the cell, tissue, organ or a portion of any of the foregoing is or comprises a cancer or tumor cell, or is a malignant tumor cell or a carcinoid tumor cell;

[0082] The cancer or tumor cell is or includes or is derived from: a malignant tumor cell (optionally a squamous cell carcinoma cell), a breast cancer cell, a colon cancer or colorectal cancer cell, a melanoma cell (optionally a malignant melanoma cell) or a multiple myeloma cell, a plasma cell, a lymphoma cell, a bladder or urothelial cancer cell, a cervical cancer cell, an ovarian cancer cell, an esophageal cancer or an esophageal squamous cell carcinoma cell; a malignant pleural mesothelioma cell, a prostate cancer cell, a cell from a microsatellite instability-high / mismatch repair-deficient tumor, a human papillomavirus-positive or Epstein-Barr-positive tumor cell, a hepatocellular carcinoma or a liver cancer cell, a lung cancer cell (optionally a non-small cell lung cancer (NSCLC)), a gastric cancer cell, a renal / kidney cancer cell, a pancreatic cancer cell, a breast cancer cell (optionally a triple-negative breast cancer cell), a lymphoma cell or a leukemia cell, or a mycosis fungoides cell; and / or

[0083] - the lymphoma or leukemia cell is or is derived from: B-cell lymphoma, acute lymphoblastic leukemia (ALL) cell, angioimmunoblastic T-cell lymphoma cell, cutaneous T-cell lymphoma, chronic myeloid leukemia in blast crisis, diffuse large B-cell lymphoma cell, hairy cell leukemia cell, follicular lymphoma cell, Burkitt's lymphoma, diffuse large B-cell lymphoma or mantle cell lymphoma, and optionally, the B-cell lymphoma is Hodgkin's lymphoma or non-Hodgkin's lymphoma, and optionally, the non-Hodgkin's lymphoma is follicular lymphoma, diffuse large B-cell lymphoma, marginal zone B-cell lymphoma, small lymphocytic lymphoma (SLL) or chronic lymphocytic leukemia (CLL), Burkitt's lymphoma or mantle cell lymphoma The invention relates to a malignant tumor cell, wherein the malignant tumor cell is a head and neck squamous cell carcinoma (HNSCC) or a basal cell carcinoma (BCC) cell.

[0084] In an alternative embodiment, a method for detecting or diagnosing cancer or a tumor is provided, wherein the method comprises detecting the expression or presence of human ICOS (inducible T-cell co-stimulatory) protein or human CD278 protein or polypeptide in or on a cell, tissue or organ sample using a method as provided herein, and wherein the specific binding of the Ab or its Ag-binding fragment or monomeric or dimeric ABP to human ICOS (inducible T-cell co-stimulatory) protein or human CD278 protein or polypeptide in or on the cell, tissue or organ or a portion of any of the foregoing is detected to detect or diagnose or assist in detecting or diagnosing the cancer or the tumor.

[0085] In an alternative embodiment of the method for detecting or diagnosing cancer or a tumor:

[0086] - the cancer or the tumor is or comprises or is derived from: a malignant tumor cell (optionally a squamous cell carcinoma cell), or a breast cancer cell, a colon cancer cell or a colorectal cancer cell, a melanoma cell (optionally a malignant melanoma cell) or a multiple myeloma cell, a plasmacytoma cell, a lymphoma cell, a bladder or urothelial cancer cell, a cervical cancer cell, an ovarian cancer cell, an esophageal cancer or an esophageal squamous cell carcinoma cell; a malignant pleural mesothelioma cell, a prostate cancer cell, a cell from a microsatellite instability-high / mismatch repair-deficient tumor, a human papillomavirus-positive or Epstein-Barr-positive tumor cell, a hepatocellular carcinoma or a liver cancer cell, a lung cancer cell (optionally a non-small cell lung cancer (NSCLC) cell), a gastric cancer cell, a renal cancer cell, a pancreatic cancer cell, a breast cancer cell (optionally a triple-negative breast cancer cell), a lymphoma cell or a leukemia cell, or a mycosis fungoides cell;

[0087] - the lymphoma or leukemia cell is or is derived from: B-cell lymphoma, acute lymphoblastic leukemia (ALL) cell, angioimmunoblastic T-cell lymphoma cell, cutaneous T-cell lymphoma, chronic myeloid leukemia in blast crisis, diffuse large B-cell lymphoma cell, hairy cell leukemia cell, follicular lymphoma cell, Burkitt lymphoma, diffuse large B-cell lymphoma or mantle cell lymphoma, and optionally, the B cell The lymphoma is Hodgkin's lymphoma or non-Hodgkin's lymphoma, and optionally the non-Hodgkin's lymphoma is follicular lymphoma, diffuse large B-cell lymphoma, marginal zone B-cell lymphoma, small lymphocytic lymphoma (SLL) or chronic lymphocytic leukemia (CLL), Burkitt's lymphoma or mantle cell lymphoma (MCL), and optionally the malignant tumor cell is a head and neck squamous cell carcinoma (HNSCC) or basal cell carcinoma (BCC) cell;

[0088] - The cell, tissue or organ sample is from an individual in need or an individual who is at high risk of developing cancer or tumor or has a family history of such cancer or tumor;

[0089] - said detecting comprises performing an immunohistochemistry (IHC) assay and optionally contacting said cell, tissue or organ sample with at least two Abs or Ag-binding fragments thereof, or monomeric or dimeric ABPs;

[0090] - at least one of the two Abs or Ag-binding fragments thereof or monomeric or dimeric ABPs comprises a variable heavy chain (VH) having an amino acid sequence comprising SEQ ID NO: 1 and a variable light chain (VL) having an amino acid sequence comprising SEQ ID NO: 2; and / or

[0091] - said at least one Ab or Ag-binding fragment thereof or monomeric or dimeric ABP comprises a heavy chain having an amino acid sequence comprising SEQ ID NO:4 and a light chain having an amino acid sequence comprising SEQ ID NO:9.

[0092] In an alternative embodiment, a method for treating, ameliorating or preventing cancer or tumors is provided, the method comprising first detecting or diagnosing the cancer or tumor using a method as provided herein, and then treating the individual in need thereof to treat, ameliorate or prevent the cancer or tumor.

[0093] In alternative embodiments of the methods for treating, ameliorating or preventing cancer or tumors as provided herein:

[0094] - the cancer or the tumor is or comprises or is derived from: a malignant tumor cell (optionally a squamous cell carcinoma cell), a breast cancer cell, a colon cancer cell or a colorectal cancer cell, a melanoma cell (optionally a malignant melanoma cell) or a multiple myeloma cell, a plasmacytoma cell, a lymphoma cell, a bladder or urothelial cancer cell, a cervical cancer cell, an ovarian cancer cell, an esophageal cancer or an esophageal squamous cell carcinoma cell; a malignant pleural mesothelioma cell, a prostate cancer cell, a cell from a microsatellite instability-high / mismatch repair-deficient tumor, a human papillomavirus-positive or Epstein-Barr-positive tumor cell, a hepatocellular carcinoma or a liver cancer cell, a lung cancer cell (optionally a non-small cell lung cancer (NSCLC)), a gastric cancer cell, a renal cancer cell, a pancreatic cancer cell, a breast cancer cell (optionally a triple-negative breast cancer cell), a lymphoma cell or a leukemia cell, or a mycosis fungoides cell;

[0095] - the lymphoma or leukemia cell is or is derived from: B-cell lymphoma, acute lymphoblastic leukemia (ALL) cell, angioimmunoblastic T-cell lymphoma cell, cutaneous T-cell lymphoma, chronic myeloid leukemia in blast crisis, diffuse large B-cell lymphoma cell, hairy cell leukemia cell, follicular lymphoma cell, Burkitt's lymphoma, diffuse large B-cell lymphoma or mantle cell lymphoma, and optionally, the B-cell lymphoma The tumor is a Hodgkin lymphoma or a non-Hodgkin lymphoma, and optionally, the non-Hodgkin lymphoma is a follicular lymphoma, a diffuse large B-cell lymphoma, a marginal zone B-cell lymphoma, a small lymphocytic lymphoma (SLL) or a chronic lymphocytic leukemia (CLL), a Burkitt lymphoma or a mantle cell lymphoma (MCL), and optionally, the malignant tumor cell is a head and neck squamous cell carcinoma (HNSCC) or a basal cell carcinoma (BCC) cell; and / or

[0096] - The cell, tissue or organ sample is from an individual in need thereof, and optionally the individual in need thereof is an individual at higher risk of developing cancer or a tumor or has a family history of said cancer or tumor.

[0097] In an alternative embodiment, provided is the use of at least one recombinant antibody (Ab) or antigen (Ag) binding fragment thereof or monomeric or dimeric antigen binding protein (ABP) as provided herein or encoded by a nucleic acid as provided herein for detecting or diagnosing cancer, or treating, ameliorating or preventing cancer.

[0098] In an alternative embodiment of the use as provided herein:

[0099] - the cancer or the tumor is or comprises or is derived from: a malignant tumor cell (optionally a squamous cell carcinoma cell), a breast cancer cell, a colon cancer cell or a colorectal cancer cell, a melanoma cell (optionally a malignant melanoma cell) or a multiple myeloma cell, a plasmacytoma cell, a lymphoma cell, a bladder or urothelial cancer cell, a cervical cancer cell, an ovarian cancer cell, an esophageal cancer or an esophageal squamous cell carcinoma cell; a malignant pleural mesothelioma cell, a prostate cancer cell, a cell from a microsatellite instability-high / mismatch repair-deficient tumor, a human papillomavirus-positive or Epstein-Barr-positive tumor cell, a hepatocellular carcinoma or a liver cancer cell, a lung cancer cell (optionally a non-small cell lung cancer (NSCLC)), a gastric cancer cell, a renal cancer cell, a pancreatic cancer cell, a breast cancer cell (optionally a triple-negative breast cancer cell), a lymphoma cell or a leukemia cell, or a mycosis fungoides cell;

[0100] - the lymphoma or leukemia cell is or is derived from: B-cell lymphoma, acute lymphoblastic leukemia (ALL) cell, angioimmunoblastic T-cell lymphoma cell, cutaneous T-cell lymphoma, chronic myeloid leukemia in blast crisis, diffuse large B-cell lymphoma cell, hairy cell leukemia cell, follicular lymphoma cell, Burkitt's lymphoma, diffuse large B-cell lymphoma or mantle cell lymphoma, and optionally, the B-cell lymphoma The tumor is a Hodgkin lymphoma or a non-Hodgkin lymphoma, and optionally, the non-Hodgkin lymphoma is a follicular lymphoma, a diffuse large B-cell lymphoma, a marginal zone B-cell lymphoma, a small lymphocytic lymphoma (SLL) or a chronic lymphocytic leukemia (CLL), a Burkitt lymphoma or a mantle cell lymphoma (MCL), and optionally, the malignant tumor cell is a head and neck squamous cell carcinoma (HNSCC) or a basal cell carcinoma (BCC) cell; and / or

[0101] - said detecting comprises performing an immunohistochemistry (IHC) assay.

[0102] In an alternative embodiment, a recombinant antibody (Ab) or antigen (Ag) binding fragment thereof or a monomeric or dimeric antigen binding protein (ABP) as provided herein is provided for use in detecting or diagnosing cancer, or treating, ameliorating or preventing cancer. In alternative embodiments, the cancer or the tumor is or includes or is derived from: a malignant tumor cell (optionally a squamous cell carcinoma cell), a breast cancer cell, a colon cancer cell or a colorectal cancer cell, a melanoma cell (optionally a malignant melanoma cell) or a multiple myeloma cell, a plasmacytoma cell, a lymphoma cell, a bladder or urothelial cancer cell, a cervical cancer cell, an ovarian cancer cell, an esophageal cancer or an esophageal squamous cell carcinoma cell; a malignant pleural mesothelioma cell, a prostate cancer cell, a cell from a microsatellite instability-high / mismatch repair-deficient tumor, a human papillomavirus-positive or Epstein-Barr-positive tumor cell, a hepatocellular carcinoma or a liver cancer cell, a lung cancer cell (optionally a non-small cell lung cancer (NSCLC)), a gastric cancer cell, a renal cancer cell, a pancreatic cancer cell, a breast cancer cell (optionally a triple-negative breast cancer cell), a lymphoma cell or a leukemia cell, or a mycosis fungoides cell. In alternative embodiments, the lymphoma or leukemia cells are or are derived from: B-cell lymphoma, acute lymphoblastic leukemia (ALL) cells, angioimmunoblastic T-cell lymphoma cells, cutaneous T-cell lymphoma, chronic myeloid leukemia in blast crisis, diffuse large B-cell lymphoma cells, hairy cell leukemia cells, follicular lymphoma cells, Burkitt's lymphoma, diffuse large B-cell lymphoma or mantle cell lymphoma, and optionally, the The B cell lymphoma is Hodgkin lymphoma or non-Hodgkin lymphoma, and optionally, the non-Hodgkin lymphoma is follicular lymphoma, diffuse large B cell lymphoma, marginal zone B cell lymphoma, small lymphocytic lymphoma (SLL) or chronic lymphocytic leukemia (CLL), Burkitt lymphoma or mantle cell lymphoma (MCL), and optionally, the malignant tumor cell is head and neck squamous cell carcinoma (HNSCC) or basal cell carcinoma (BCC) cell. In an alternative embodiment, the detection or diagnosis includes performing an immunohistochemistry (IHC) assay,

[0103] In alternative embodiments, a kit is provided, the kit comprising: a chimeric or recombinant antibody as provided herein; a chimeric or recombinant nucleic acid as provided herein; or an expression cassette, vector, recombinant virus, artificial chromosome, cosmid or plasmid as provided herein; or a cell as provided herein. In alternative embodiments of the kit as provided herein, the kit comprises components required for immunohistochemistry (IHC) assays, and / or comprises instructions for practicing the methods as provided herein.

[0104] The details of one or more exemplary embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

[0105] All publications, patents, and patent applications cited herein are expressly incorporated by reference in their entirety for all purposes. BRIEF DESCRIPTION OF THE DRAWINGS

[0106] The patent or application file contains at least one drawing drawn in color. Copies of the patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0107] The drawings set forth herein are illustrative of exemplary embodiments provided herein and are not meant to limit the scope of the present invention as encompassed by the claims.

[0108] Figures 1A to 1D The following images are shown: IHC comparing an exemplary anti-human ICOS antibody (Ab) as provided herein (the Ab is also referred to as T0251A) with a reference anti-human ICOS antibody Ab (Abscam, referred to as SP98) in IHC staining of the following cells, the exemplary anti-human ICOS antibody having: a heavy chain having an amino acid sequence comprising SEQ ID NO: 4; and a light chain having an amino acid sequence comprising SEQ ID NO: 9:

[0109] - Tonsillar cells ( Figure 1A )、T0251A upper figure, SP98 lower figure;

[0110] -Melanoma cells ( Figure 1B )、T0251A upper figure, SP98 lower figure;

[0111] -Hepatocytes ( Figure 1C ), T0251A upper figure, SP98 lower figure; and

[0112] -Colonocytes ( Figure 1D ), T0251A upper figure, SP98 lower figure.

[0113] The same reference symbols in the various drawings denote the same components. DETAILED DESCRIPTION

[0114] In alternative embodiments, chimeric, synthetic or recombinant anti-human ICOS protein (also known as: inducible T cell co-stimulator, or cluster of differentiation-278 or CD278; activation-induced lymphocyte immunomodulatory molecule; AILIM; and CVID1) binding polypeptides are provided, including ICOS binding antibodies (Abs), including manufactured products and kits containing them; and methods for making and using them, including, for example, their use in the detection or diagnosis and treatment of cancer or other diseases or disorders involving ICOS expression.

[0115] Although the present invention is not limited to any particular mechanism of action, in alternative embodiments, an antibody or antigen binding protein as provided herein targets (and specifically binds to) an ICOS protein expressed on the surface of a cancer cell, such as seen in some types of cancer, such as malignant tumors (optionally squamous cell carcinoma), breast cancer, colon cancer or colorectal cancer, melanoma (optionally malignant melanoma) or multiple myeloma, plasmacytoma, lymphoma, bladder cancer or urothelial carcinoma, cervical cancer, ovarian cancer, esophageal cancer or esophageal squamous cell carcinoma; malignant pleural mesothelioma, prostate cancer, microsatellite instability high / mismatch repair deficient tumors, human papillomavirus-positive or Epstein-Barr-positive tumors, hepatocellular carcinoma or liver cancer, lung cancer (optionally non-small cell lung cancer (NSCLC)), gastric cancer, kidney cancer, pancreatic cancer, breast cancer (optionally triple-negative breast cancer), lymphoma or leukemia, or mycosis fungoides.

[0116] Expression of recombinant or chimeric antibodies

[0117] In alternative embodiments, chimeric or recombinant antibodies as provided herein, including exemplary chimeric or recombinant anti-human ICOS antibodies, with or without a signal peptide, can be expressed as recombinant Abs using plasmids (or any expression vector) encoding the corresponding heavy and light chains, or the heavy and light chains can be encoded in separate expression vectors.

[0118] In some embodiments, the heavy and light chains can be grown in HEK293-6E cells from pTT5 TM Vector (National Research Council Canada, NRC-CNRC, Canada) (cis or trans) expression. In alternative embodiments, for example, in a stable cell line capable of synthesizing, optionally inducibly synthesizing, the vector expressing the heavy chain and / or light chain is additional or chromosomally integrated.

[0119] In alternative embodiments, the nucleic acid of chimeric or recombinant Ab as provided herein is provided. Can be prepared, separated and / or manipulated by cloning and expression of cDNA library, courier or genomic DNA pcr amplification etc. as provided herein. Be used for putting into practice the nucleic acid of the embodiment as provided herein, no matter it is RNA, cDNA, genomic DNA, carrier, virus or their heterozygote, can separate from multiple sources, through genetic engineering, amplification and / or recombinantly express / produce. Can separate or clone individually from the recombinant polypeptide produced by these nucleic acids, and test required activity. Can use any recombinant expression system, comprise bacterium, fungi, mammals, yeast, insect or plant cell expression system.

[0120] Alternatively, these nucleic acids can be synthesized in vitro by well-known chemical synthesis techniques, as described, for example, in Adams (1983) J. Am. Chem. Soc. 105:661; Belousov (1997) Nucleic Acids Res. 25:3440-3444; Frenkel (1995) Free Radic. Biol. Med. 19:373-380; Blommers (1994) Biochemistry 33:7886-7896; Narang (1979) Meth. Enzymol. 68:90; Brown (1979) Meth. Enzymol. 68:109; Beaucage (1981) Tetra. Lett. 22:1859; U.S. Pat. No. 4,458,066.

[0121] Techniques for nucleic acid manipulation, such as subcloning, labeling probes (e.g., random primer labeling using Klenow polymerase, nick translation, amplification), sequencing, hybridization, and the like are well described in the scientific and patent literature, see, for example, Sambrook, ed., MOLECULAR CLONING: A LABORATORY MANUAL (2nd ed.), Vols. 1-3, Cold Spring Harbor Laboratory, (1989); CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, Ausubel, ed., John Wiley & Sons, Inc., New York (1997); LABORATORY TECHNIQUES IN BIOCHEMISTRY AND MOLECULAR BIOLOGY: HYBRIDIZATION WITH NUCLEIC ACID PROBES, Part I. Theory and Nucleic Acid Preparation, Tijssen, ed. Elsevier, NY (1993).

[0122] Another useful means of obtaining and manipulating nucleic acids for practicing the embodiments as provided herein includes screening and recloning insert sequences isolated or amplified from, for example, genomic clones or cDNA clones. Nucleic acid sources include recombinant nucleic acid sequences, genomic or cDNA libraries contained in and / or expressed in, for example, mammalian artificial chromosomes (MACs), see, for example, U.S. Patent Nos. 5,721,118; 6,025,155; human artificial chromosomes, see, for example, Rosenfeld (1997) Nat. Genet. 15: 333-335; yeast artificial chromosomes (YACs); bacterial artificial chromosomes (BACs); P1 artificial chromosomes, see, for example, Woon (1998) Genomics 50: 306-316; P1-derived vectors (PACs), see, for example, Kern (1997) Biotechniques 23: 120-124; cosmids, recombinant viruses, phages, phagemids or plasmids.

[0123] In alternative embodiments, nucleic acids as provided herein are operably linked to transcriptional regulatory elements, including a promoter, which may be constitutive or inducible transcriptional regulatory elements.

[0124] In alternative aspects, an "expression cassette" comprising a nucleotide sequence as provided herein is provided, the nucleotide sequence for example encoding a chimeric or recombinant antibody as provided herein. The expression cassette may include at least one transcriptional regulatory element, such as a promoter, which is operably connected to the antibody coding sequence and may optionally also include a transcription termination signal. Additional factors necessary for or conducive to affecting expression, such as enhancers, may also be used.

[0125] In alternative aspects, the expression cassette for practicing the embodiments as provided herein includes plasmids, expression vectors, recombinant viruses, any form of recombinant "naked DNA" vectors, etc. In alternative aspects, the "vector" for practicing the embodiments as provided herein may include nucleic acids that can infect, transfect, transiently or permanently transduce cells. In alternative aspects, the vector for practicing the embodiments as provided herein may be naked nucleic acid, or a nucleic acid compounded with protein or lipid. In alternative aspects, the vector for practicing the embodiments as provided herein may include viral or bacterial nucleic acids and / or proteins, and / or membranes (e.g., cell membranes, viral lipid envelopes, etc.). In alternative aspects, the vector for practicing the embodiments as provided herein may include but is not limited to replicons (e.g., RNA replicons, bacteriophages), and DNA fragments may be attached to the replicons and replicated. Therefore, vectors include but are not limited to RNA, autonomous self-replicating circular or linear DNA or RNA (e.g., plasmids, viruses, etc., see, e.g., U.S. Patent No. 5,217,879), and may include both expression plasmids and non-expression plasmids. In alternative aspects, vectors used to practice embodiments as provided herein can be stably replicated by the cell during mitosis as an autonomous structure, or can be integrated within the host genome.

[0126] In alternative aspects, "promoter" for practicing the embodiments provided herein includes all sequences that can drive the coding sequence to transcribe in cells, such as bacteria, yeast, fungi, plants, insects (e.g., baculovirus) or mammalian cells. Therefore, the promoter used in the construct includes cis-acting transcription control elements and regulatory sequences, and the cis-acting transcription control elements and regulatory sequences participate in the timing and / or rate of regulating or regulating gene transcription. For example, the promoter for practicing the embodiments provided herein can be a cis-acting transcription control element, including enhancers, promoters, transcription terminators, replication origins, chromosome integration sequences, 5' and 3' non-translated regions or intron sequences, and the transcription control elements participate in transcriptional regulation. These cis-acting sequences can interact with proteins or other biomolecules to perform (on / off, regulation, regulation, etc.) transcription.

[0127] " constitutive " promoter that is used to put into practice the embodiment that provides as this paper can be the promoter that drives expression continuously under most environmental conditions and development or cell differentiation state. " inducible " or " regulatable " promoter that is used to put into practice the embodiment that provides as this paper can instruct the expression of nucleic acid that provides as this paper under the influence of environmental conditions or developmental conditions. The example that can influence the environmental conditions that is transcribed by the inducible promoter that is used to put into practice the embodiment that provides as this paper comprises the existence of the inducing factor that is applied to cell.

[0128] In alternative embodiments, the nucleic acid used to practice the embodiments as provided herein encodes a polypeptide having the following amino acid sequence:

[0129] Summary of Exemplary Polypeptide Sequences

[0130] (SEQ ID NO: 1) Variable domain of IgG heavy chain

[0131] (SEQ ID NO:2) Variable domain of kappa 1 light chain

[0132] (SEQ ID NO:3) Heavy chain constant region:

[0133] (SEQ ID NO:4) Heavy chain variable region and constant region:

[0134] (SEQ ID NO:5) Heavy chain variable region amino acid terminal signal sequence

[0135] (SEQ ID NO:6) Heavy chain variable region with signal sequence

[0136] (SEQ ID NO: 7) A heavy chain having variable and constant domains and having an amino-terminal signal sequence

[0137] (SEQ ID NO: 8) Light chain constant region

[0138] (SEQ ID NO:9) Light chain with variable region and constant region

[0139] (SEQ ID NO: 10) Light chain variable domain amino terminal signal sequence

[0140] (SEQ ID NO: 11) Light chain variable domain with signal sequence

[0141] (SEQ ID NO: 12) Light chain with signal sequence

[0142] Exemplary polypeptide sequences

[0143] SEQ ID NO: 1 (variable region of IgG heavy chain)

[0144] QSLEESGGRLVTPGTPLTLTCTVS GFSLSSYG VSWVRQAPGKGLEWIGI INSDHST YYAKWAKGRFTISKTSTTVDLKITSPTTEDTATYFC ARSYGIGSIF WGPGTLVTVSS

[0145] SEQ ID NO:2 (Variable domain of κ1 light chain)

[0146] AAVLTQTPSPVSAAVGGTVSISCQSS KSVYNNNQ LSWFQQKPGQRPKLLIY EAF KLPSGVPSRFKGSGSGTQFTLTISDVQCDDAATYYC AAVYSDDSDNS FGGGTEVVVK

[0147] (SEQ ID NO:3) Heavy chain constant region:

[0148] GQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVTVTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSVTSSSQPVTCNVAHPATNTKVDKTVAPSTCSKPTCPPPELLGGPSVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPAPIEKTISKARGQPLEPKVYTMGPPREELSSRSVSLTCMINGFYPSDISVEWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSVPTSEWQRGDVFTCSVMHEALHNHYTQKSISRSPGK

[0149] (SEQ ID NO:4) Heavy chain variable and constant regions:

[0150] QSLEESGGRLVTPGTPLTLTCTVSGFSLSSYGVSWVRQAPGKGLEWIGIINSDHSTYYAKGRFTISKTSTTVDLKITSPTTEDTATYFCARSYGIGSIFWGPGTLV TVSSGQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVTVTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSVTSSSQPVTCNVAHPATNTKVDKTVAPSTCSKPTC PPPELLGGPSVFIFPPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPAPIEKTISK ARGQPLEPKVYTMGPPREELSSRSVSLTCMINGFYPSDISVEWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSVPTSEWQRGDVFTCSVMHEALHNHYTQKSISRSPGK

[0151] (SEQ ID NO:5) Heavy chain variable region amino acid terminal signal sequence

[0152] METGLRWLLLVAVLKGVQC

[0153] (SEQ ID NO:6) Heavy chain variable region with signal sequence

[0154] METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLSSYGVSWVRQAPGKGLEWIGIINSDHSTYYAKWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARSYGIGSIFWGPGTLVTVSS

[0155] Heavy chain having variable and constant domains and having an amino-terminal signal sequence

[0156] (SEQ ID NO:7)METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLSSYGVSWVRQAPGKGLEWIGIINSDHSTYYAKWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARSYGIGSIFWGPGTLVTVSSGQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVTVTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSVTSSSQPVTCNVAHPATNTKVDKTVAPSTCSKPTCPPPELLGGPSVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPAPIEKTISKARGQPLEPKVYTMGPPREELSSRSVSLTCMINGFYPSDISVEWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSVPTSEWQRGDVFTCSVMHEALHNHYTQKSISRSPGK

[0157] Light chain constant region:

[0158] (SEQ ID NO:8)GDPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTVTWEVDGTTQTTGIENSKTPQNSADCTYNLSSTLTLTSTQYNSHKEYTCKVTQGTTSVVQSFNRGDC

[0159] Light chain with variable and constant regions:

[0160] (SEQ ID NO:9)AAVLTQTPSPVSAAVGGTVSISCQSSKSVYNNNQLSWFQQKPGQRPKLLIYEAFKLPSGVPSRFKGSGSGTQFTLTISDVQCDDAATYYCAAVYSDDSDNSFGGGTEVVVKGDPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTVTWEVDGTTQTTGIENSKTPQNSADCTYNLSSTLTLTSTQYNSHKEYTCKVTQGTTSVVQSFNRGDC

[0161] (SEQ ID NO: 10) Light chain variable domain amino terminal signal sequence MDTRAPTQLLGLLLLWLPGATF

[0162] (SEQ ID NO: 11) The light chain variable domain with a signal sequence comprises

[0163] MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVSISCQSSKSVYNNNQLSWFQQKPGQRPKLLIYEAFKLPSGVPSRFKGSGSGTQFTLTISDVQCDDAATYYCAAVYSDDSDNSFGGGTEVVVK

[0164] (SEQ ID NO: 12) The light chain with a signal sequence comprises

[0165] MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVSISCQSSKSVYNNNQLSWFQQKPGQRPKLLIYEAFKLPSGVPSRFKGSGSGTQFTLTISDVQCDDAATYYCAAVYSD DSDNSFGGGTEVVVKGDPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTVTWEVDGTTQTTGIENSKTPQNSADCTYNLSSTLTLTSTQYNSHKEYTCKVTQGTTSVVQSFNRGDC

[0166] Peptides, polypeptides, peptide mimetics

[0167] In alternative embodiments, the peptides and polypeptides used to practice the embodiments as provided herein may comprise any "mimetic" and / or "peptide mimetic" form. In alternative embodiments, the peptides and polypeptides used to practice the embodiments as provided herein may comprise synthetic chemical compounds having substantially the same structural and / or functional characteristics as natural polypeptides, such as chimeric or recombinant antibodies as provided herein. The mimetic used to practice the embodiments as provided herein may be composed entirely of synthetic non-natural amino acid analogs, or may be a chimeric molecule of a portion of natural peptide amino acids and a portion of non-natural amino acid analogs. The mimetic may also incorporate any amount of natural amino acid conservative substitutions, as long as such substitutions do not substantially change the structure and / or activity of the mimetic. Routine experiments will determine whether the mimetic is effective in practicing the present invention, for example, whether the mimetic composition is effective in specifically binding to the ICOS protein. The methodology described in detail herein and other methodologies known to those skilled in the art can be used to select or guide people to select effective mimetic to practice the compositions and / or methods of the present invention.

[0168] The polypeptide mimetic composition for practicing the embodiments as provided herein can include any combination of non-natural structural components. In alternative aspects, the mimetic composition for practicing the embodiments as provided herein can include one or all of the following three structural groups: a) residue linking groups other than natural amide bonds ("peptide bonds"); b) non-natural residues that replace naturally occurring amino acid residues; or c) residues that induce secondary structure simulation, i.e., induce or stabilize secondary structure, such as β turns, γ turns, β folds, α helical conformations, etc. For example, when all or part of the residues of a polypeptide are joined by chemical means other than non-natural peptide bonds, the polypeptide can be characterized as a mimetic. In alternative aspects, the mimetic composition for practicing the embodiments as provided herein can be a peptoid, in which the side chain is connected to the nitrogen of the peptide backbone, rather than to the α-carbon as in a peptide; and the peptoid lacks the amide hydrogens responsible for many of the secondary structural elements in peptides and proteins. In alternative aspects, the mimetic composition for practicing the embodiments as provided herein can be a beta peptide (β peptide), in which the amino group is attached to the β-carbon (i.e., the carbon two atoms away from the carboxylate group) in the β peptide.

[0169] In alternative embodiments, peptides and polypeptides for practicing embodiments as provided herein can be intentionally deuterated, e.g., hydrogen at a specific position is replaced by deuterium ("D"), and it should be understood that the abundance of deuterium at the position is greater than or significantly greater than the natural abundance of deuterium (which is 0.015%). For example, in alternative embodiments, deuterium substitution or "enrichment" occurs at one or more specific positions. In one embodiment, the deuterium enrichment is not less than about 1%, or the deuterium enrichment is not less than about 1%, 5%, 10%, 20%, 50%, 70%, 80% or 90%, or between about 1% and 99%.

[0170] Exemplary Immunohistochemistry (IHC) Techniques and Complete Equipment

[0171] In alternative embodiments, immunohistochemistry (IHC) methodology and / or reagents for practicing compositions, articles of manufacture, kits or methods as provided herein may include or comprise the use of any IHC protocol, IHC repertoire, apparatus and / or image or data analysis system for practicing IHC or IHC reagents known in the art, such as described in: U.S. Patent No. (USPN) 11,321,881 (describing IHC imaging protocols and apparatus); 11,249,085, which describes the use of click chemistry for signal amplification in IHC and ISH assays; 11,222,424 (describing IHC imaging apparatus and computer programs); 11,143,648 (describing IHC imaging apparatus and computer programs); , which describes new pigments for chromogenic IHC and ISH staining using multi-dye quinone methides and tyramine conjugates; 11,047,774 (which describes an automated IHC specimen processing system); 11,028,044, which describes methods for commercial-scale preparation of 3,3'5,5'-tetramethylbenzidine (TMB) and its salts; 10,977,791, which describes computer-implemented methods for analyzing tissue samples; 10,948,493, which describes IHC methods; 10,937,162 (which describes an IHC image analysis algorithm); 10,816,443, which describes an automated batch stainer for staining biological specimens on microscope slides; 1 0,718,773, which describes a method for determining whether a subject with a malignant tumor is eligible for treatment with a therapeutic agent; 10,565,479 (which describes a method for identifying blurred areas in a digital image of stained tissue); 10,564,076 (which describes a system for sample preparation for analysis (or IHC); 10,551,395 (which describes an automated histological staining system); 10,551,378 (which describes a tissue staining method); 10,504,224 (which describes a digital tissue image analysis system for IHC); 10,501,777 (which describes simultaneous, multiplexed detection and quantification of protein expression in IHC); 10,488, 340 (describing methods for extracting images of target fluorophores in biological materials); 10,453,195 (describing methods for detecting tissue regions of interest using digital pathology imaging); 10,438,381 (describing devices, systems, and methods for generating digital images of tissue sections); 10,416,176 (describing methods for processing specimens in automated histological staining systems); 10,393,633 (describing methods for processing and inhibiting degradation of IHC samples); 10,217,011 (describing the disposal of IHC slides); 10,209,165 (describing automated or semi-automated methods for assessing the staining quality of specimens containing cells);10,126,216 (describing methods for fixing tissue samples for IHC); 9,423,322; 9,103,822, describing polymer supports for immunohistochemistry and in situ hybridization; 8,515,683 (describing methods and systems for automated detection of immunohistochemistry (IHC) patterns); USPN 10,816,443 (describing an automated batch stainer for staining biological specimens on microscope slides); or as described in: U.S. Patent Application Publication No. US2021 / 0239683A1, which describes a composition for forming a porous hydrogel around cells, the composition being suitable for immunostaining cells within the hydrogel; or USPN 11,112,413, or US2022 / 0057408 A1, which describes a method for detecting one or more targets in an IHC tissue sample using an epitope-tagged antibody; or US2021 / 0048432 A1, which describes direct immunohistochemistry (IHC) staining and direct immunocytochemistry (ICC) techniques; or US2020 / 0292536, which describes synthetic controls that can be used in immunohistochemistry (IHC) assays; or US2022 / 0057408 A1, which describes the use of epitope-tagged antibodies in immunohistochemistry (IHC) assays; or US2021 / 0201485 A1, which describes a computer-implemented method for analyzing tissue samples; US2019 / 0178867 A1 (describing the detection of specific tissue objects within thin sections of tissue samples as imaged under a brightfield microscope without the use of chromogenic dyes specific to these tissue objects); US2019 / 0156510 A1 (describing an image analysis method for analyzing IHC tissue samples); US2019 / 0293637 A1 (methods and systems for quantitative immunohistochemistry (IHC) of target protein molecules); US2019 / 0080450 A1 (describing automated determination of staining quality of biological specimens stained by IHC); or US 2020 / 0316589A1 (describing a porous solid support container for processing and testing of fixed biological materials); or US2022 / 0229062 A1, which describes a method for producing rapid immunohistochemical detection of antigens from biological specimens; US2021 / 0371520 A1, which describes various immunohistochemical (IHC) methodologies; or, US2022 / 0034766 A1, which describes immunohistochemical staining techniques for identifying cells. ;

[0172] In alternative embodiments, the chimeric or recombinant antibody, antigen-binding fragment thereof, or monomeric or dimeric antigen-binding protein in an IHC protocol or kit as provided herein is substantially purified or isolated, or is in the form of an unpurified or partially purified culture supernatant.

[0173] In alternative embodiments, methods as provided herein may use or comprise reagents for detecting or visualizing antibody-antigen interactions using, for example, any product or method, and IHC protocols or reagents known in the art.

[0174] In alternative embodiments, the methods as provided herein include the use of chromogenic immunohistochemistry (CIH), wherein a primary antibody (e.g., a chimeric or recombinant antibody (Ab) or antigen-binding fragment thereof, or a monomeric or dimeric antigen-binding protein as provided herein) or a secondary antibody (e.g., wherein the secondary antibody binds (the primary antibody) to the chimeric or recombinant antibody or antigen-binding fragment thereof, or the monomeric or dimeric antigen-binding protein after the chimeric or recombinant antibody (Ab) or antigen-binding fragment thereof, or the monomeric or dimeric antigen-binding protein as provided herein has specifically bound, paired or associated with an ICOS epitope or polypeptide) is conjugated to an enzyme that can catalyze a reaction that produces a color, such as a peroxidase (or immunoperoxidase), such as horseradish peroxidase (HRP).

[0175] In alternative embodiments, the methods as provided herein include the use of immunofluorescence, wherein the primary or secondary antibody is labeled with a fluorophore (e.g., fluorescein or fluorescein isothiocyanate (FITC)), a triarylmethane dye, such as rhodamine or a rhodamine derivative (e.g., tetramethylrhodamine (TRITC), rhodamine 6G, rhodamine 123, rhodamine B, carboxytetramethylrhodamine (TAMRA), tetramethylrhodamine (TMR), sulforhodamine 101)), aminomethylcoumarin acetate (AMCA), Alexa Fluor®, TM or Dylight TM Fluorescent dyes. 3,3'-Diaminobenzidine (DAB) can also be used.

[0176] In alternative embodiments, methods as provided herein include the use of a direct method or a one-step staining method, wherein, for example, in a tissue section, a primary antibody (e.g., a chimeric or recombinant antibody (Ab) or an antigen-binding fragment thereof, or a monomeric or dimeric antigen-binding protein as provided herein) is labeled and reacts directly with the antigen. Although this technique utilizes only one antibody and is therefore simple and rapid, the sensitivity may be low because the signal amplification is very small.

[0177] In alternative embodiments, methods as provided herein include the use of an indirect method in which an unlabeled primary antibody (first layer) binds to a target antigen (e.g., TTF-1) in, for example, a tissue or organ, and then a labeled secondary antibody (second layer) reacts with the primary antibody. The secondary antibody can be directed against an isotype, e.g., IgG of the animal species from which the primary antibody is derived. This method may be more sensitive than a direct detection strategy because if several secondary antibodies are conjugated to a detection reagent such as a fluorescent or enzyme reporter molecule, the signal amplification caused by the secondary antibody binding to each primary antibody.

[0178] In an alternative embodiment, if the secondary antibody is conjugated to several detection molecules, such as a biotin molecule, the biotin molecule can recruit avidin-, streptavidin- or NeutrAvidin TM The protein-binding enzyme complex achieves further amplification.

[0179] In alternative embodiments, IHC is performed on tissue sections or tissue biopsy sections (e.g., paraformaldehyde (PFA) fixed tissue or organ, or formalin fixed paraffin embedded tissue). In alternative embodiments, the tissue is sliced ​​or used as a whole. Before slicing, the tissue sample can be embedded in a medium, such as paraffin or freezing medium. Tissue sections can be sliced ​​on a variety of instruments, most commonly microtomes, cryostat slicers, or vibrating slicers. The specimen can be sliced ​​in the range of about 3 μm to 5 μm. The sections can be mounted on slides, dehydrated using increasing concentrations of alcohol washes (e.g., 50%, 75%, 90%, 95%, 100%), and cleaned with a detergent such as xylene, and then imaged under a microscope.

[0180] Depending on the method of fixation and tissue preservation, samples may require additional steps to make the ICOS epitope available for antibody binding, including dewaxing and antigen retrieval. For formalin-fixed paraffin-embedded tissues, antigen retrieval is often necessary and may include pre-treatment of sections with heat or proteases.

[0181] In an alternative embodiment, the EnVision DuoFLEX Doublestain System is used TM IHC was performed using the EnVision DuoFLEX Dual Staining System (Agilent, San Jose, CA), which allows staining of two or more markers on a single slide. In an alternative embodiment, EnVision FLEX HRP Magenta High pH (DakoOmnis TM , Agilent, San Jose, CA) system to perform IHC, and the combination can be performed by EnVision FLEXHRP TMMagenta chromogen visualization. In an alternative embodiment, the EnVision FLEX micro kit is used TM To perform IHC at high pH, ​​the EnVision FLEX Micro Kit High pH is a high-sensitivity visualization system designed to work with the DAKOAutostainer TM The instruments are used together in IHC; this dual-link system detects mouse and rabbit primary antibodies, and the reaction is visualized by 3,3'-diaminobenzidine (DAB) chromogen (DAB forms a water-insoluble brown precipitate when oxidized, for example, by peroxidase).

[0182] Manufacturing products and kits

[0183] Provided are articles of manufacture and kits for practicing the methods as provided herein, e.g., comprising a chimeric or recombinant anti-ICOS binding protein, e.g., an anti-ICOS polypeptide Ab, as provided herein; and optionally, the articles of manufacture and kits may further comprise some or all of the reagents required to perform IHC, and optionally may comprise instructions for practicing the methods as provided herein.

[0184] In alternative embodiments, the article of manufacture or kit comprises a mixture or cocktail of antibodies (Ab) as provided herein, e.g., a mixture or cocktail comprising two, three or more anti-human ICOS binding proteins (e.g., anti-ICOS antibodies (Ab)).

[0185] In alternative embodiments, the article of manufacture or kit comprises a mixture or cocktail of antibodies (Ab), including antibodies comprising the heavy chain and / or light chain CDRs of an antibody as provided herein, or antibodies produced by an antibody-producing clone as provided herein.

[0186] In alternative embodiments, the article of manufacture or kit comprises an antibody producing clone as provided herein.

[0187] Any of the above aspects and embodiments may be combined with any other aspects or embodiments disclosed herein in the Summary, Figures, and / or Detailed Description sections.

[0188] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0189] Unless otherwise stated or obvious from the context, as used herein, the term "or" is construed as inclusive and encompasses both "or" and "and."

[0190] Unless otherwise specified or apparent from the context, as used herein, the term "about" is understood to be within the normal tolerance range of the art, such as within 2 standard deviations of the mean. About (the use of the term "about") can be understood to be within 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05% or 0.01% of the stated value. Unless otherwise clearly stated from the context, all numerical values ​​provided herein are modified by the term "about".

[0191] Unless specifically stated or obvious from the context, as used herein, the terms "substantially all," "substantially a majority," "substantially all," or "majority" encompass at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% or more of a reference amount of a composition.

[0192] The entire contents of each patent, patent application, publication and document cited herein are hereby incorporated by reference. The reference to the above patents, patent applications, publications and documents does not admit that any of the above is relevant prior art, nor does it constitute any admission of the contents or dates of these publications or documents. The reference to these documents alone should not be construed as an assertion or admission that any part of the contents of any document is considered to be necessary material to satisfy the statutory disclosure requirements of patent applications in any country or region. Nevertheless, the right to rely on any such document to provide materials that the examining body or court considers to be essential to the subject matter claimed is reserved where appropriate.

[0193] Without departing from the basic aspects of the present invention, the foregoing may be modified. Although the present invention has been described in considerable detail with reference to one or more specific embodiments, it will be appreciated by those of ordinary skill in the art that changes may be made to the embodiments specifically disclosed in this application, and these modifications and improvements are still within the scope and spirit of the present invention. The present invention illustratively described herein may be properly practiced in the absence of any elements not specifically disclosed herein. Thus, for example, in each instance herein, any of the terms "comprising," "consisting essentially of," and "consisting of" may be replaced by any of the other two terms. Therefore, the terms and expressions that have been used are used as terms of description rather than limitation, and equivalents of the features shown and described or parts thereof are not excluded, and it should be recognized that various modifications are possible within the scope of the present invention. Embodiments of the present invention are set forth in the following claims.

[0194] The present invention will be further described with reference to the examples described herein; however, it should be understood that the invention is not limited to such examples.

[0195] Example

[0196] Unless otherwise indicated in the examples, all recombinant DNA techniques were performed according to standard protocols, for example as described in Sambrook et al., (2012) Molecular Cloning: A Laboratory Manual, 4th edition, Cold Spring Harbor Laboratory Press, NY and in Ausubel et al., (1994) Current Protocols in Molecular Biology, Current Protocols, USA, Volumes 1 and 2. Other references to standard molecular biology techniques include Sambrook and Russell (2001) Molecular Cloning: A Laboratory Manual, 3rd edition, Cold Spring Harbor Laboratory Press, NY; Brown (1998) Molecular Biology Lab Fax, 2nd edition, Volumes I and II of Academic Press (UK). Standard materials and methods for polymerase chain reaction can be found in Dieffenbach and Dveksler (1995) PCR Primer: A Laboratory Manual, Cold Spring Harbor Laboratory Press, and in McPherson et al., (2000) PCR-Basics: From Background to Bench, 1st edition, Springer Verlag, Germany.

[0197] Example 1: Preparation of exemplary Ab

[0198] ICOS antigen was designed using part of the intracellular domain of human ICOS. The antigen was produced as a synthetic peptide. This antigen was used to immunize rabbits and blood was subsequently collected to confirm serum reactivity to human ICOS in ELISA and IHC.

[0199] Rabbits showing the best immune response to human ICOS tested on various ICOS positive human tissues and different non-expressing tissues were selected for B cell selection using blood samples from the immunized rabbits.

[0200] Briefly, B cells expressing antibodies that bind to the immunogen are isolated as monoclones, cultured, and then tested for ICOS specificity in ELISA. ELISA-specific clones are further tested in ultrasensitive IHC on normal and clinical tissues using a high pH antigen retrieval buffer. The best performing clones are selected based on IHC performance. The antibody variable domains are cloned into pTT5-based TM The recombinant antibodies were expressed in HEK293-6E cells in a custom expression vector containing the constant regions of the heavy chain and the κ1 light chain (National Research Council Canada, NRC-CNRC, Canada).

[0201] Recombinant antibodies were tested for human ICOS binding by biolayer interferometry (BLI) on BLItz followed by IHC on normal and clinical tissues by standard FLEX protocols.

[0202] Some antibodies were identified as showing ICOS specific staining in IHC. Clone 1E9 / 4B7 showed very promising results on the tissues tested. The specificity of the antibody has been tested on different clinical tissues, showing its usability for in vitro diagnostics using immunohistochemistry.

[0203] Figures 1A to 1D The following images are shown: In tonsil cells ( Figure 1A ), melanoma cells ( Figure 1B ), liver cells ( Figure 1C ) and colon cells ( Figure 1D ), as provided herein, an exemplary anti-human ICOS antibody (Ab) (also referred to as T0251A) is compared to a reference anti-human ICOS Ab (Abscam, referred to as SP98), wherein the exemplary anti-human ICOS antibody has: a heavy chain having an amino acid sequence comprising SEQ ID NO: 4; and a light chain having an amino acid sequence comprising SEQ ID NO: 9. IHC staining of T0251A and SP98 was performed using DAKO OMNIS TM (Agilent, San Jose, CA) staining IHC protocol. T0251A was used at 0.5 μg / mL and SP98 was used at a 1 / 50 dilution. High pH target retrieval solution containing rabbit linker was used for IHC of both T0251A and SP98. TM (Target Retrieval Solution TM , TRS) (Agilent, San Jose, CA).

[0204] A number of embodiments of the present invention have been described. However, it will be appreciated that various modifications may be made without departing from the spirit and scope of the present invention. Therefore, other embodiments are within the scope of the claims.

Claims

1. An isolated or purified antibody (Ab) or antigen (Ag) binding fragment thereof, or a monomeric or dimeric antigen binding protein (ABP), which can specifically bind to human ICOS (inducible T cell co-stimulatory) protein or human CD278 polypeptide, The isolated or purified Ab or its Ag binding fragment, or monomer or dimer ABP comprises: (a) a heavy chain variable region (VH), said VH comprising: (1) an amino acid sequence comprising the three CDR1, CDR2 and CDR3 complementarity determining regions (CDRs) of SEQ ID NO: 1, or CDR1 amino acid (aa) residues GFSLSSYG (residues 25 to 32 of SEQ ID NO: 1), CDR2 aa residues INSDHST (residues 50 to 56 of SEQ ID NO: 1), and CDR3 aa residues ARSYGIGSIF (residues 93 to 102 of SEQ ID NO: 1), or (2) an amino acid sequence having at least about 70%, 75%, 80%, 85%, 90%, 95%, 98% sequence identity, or between about 70% and 100% sequence identity to each of the three CDR1, CDR2, and CDR3 complementarity determining regions (CDRs) of SEQ ID NO: 1, or CDR1 amino acid (aa) residues GFSLSSYG (residues 25-32 of SEQ ID NO: 1), CDR2 aa residues INSDHST (residues 50 to 56 of SEQ ID NO: 1), and CDR3 aa residues ARSYGIGSIF (residues 93-102 of SEQ ID NO: 1), or (3) an amino acid sequence having at least about 70%, 75%, 80%, 85%, 90%, 95%, 98% sequence identity, or between about 70% and 100% sequence identity to SEQ ID NO: 1, or an amino acid sequence having complete sequence identity to SEQ ID NO: 1; or (b) a light chain variable region (VL), said VL comprising: (1) an amino acid sequence comprising the three CDR1, CDR2 and CDR3 complementarity determining regions (CDRs) of SEQ ID NO: 2, or CDR1 amino acid (aa) residues KSVYNNNQ (residues 27 to 34 of SEQ ID NO: 2), CDR2 aa residues EAF (residues 52 to 54 of SEQ ID NO: 2), and CDR3 aa residues AAVYSDDSDNS (residues 91 to 101 of SEQ ID NO: 2), or (2) an amino acid sequence having at least about 70%, 75%, 80%, 85%, 90%, 95%, 98% sequence identity, or between about 70% and 100% sequence identity to each of the three CDR1, CDR2, and CDR3 complementarity determining regions (CDRs) of SEQ ID NO:2, or CDR1 amino acid (aa) residues KSVYNNNQ (residues 27-34 of SEQ ID NO:2), CDR2 aa residues EAF (residues 52 to 54 of SEQ ID NO:2), and CDR3 aa residues AAVYSDDSDNS (residues 91-101 of SEQ ID NO:2), or (3) an amino acid sequence having at least about 70%, 75%, 80%, 85%, 90%, 95%, 98% sequence identity, or between about 70% and 100% sequence identity to SEQ ID NO: 2, or an amino acid sequence having complete (100%) sequence identity to SEQ ID NO: 2; or (c) the heavy chain variable region (VH) of (a) and the light chain variable region (VL) of (b).

2. The isolated or purified Ab or its Ag binding fragment, or monomer or dimer ABP according to claim 1, which is manufactured or is in the following form: Antigen binding fragment (Fab, or Ab fragment having only one constant domain and one variable domain of each of the Ab heavy and light chains), F(ab')2 (or Ab digested by pepsin to produce the following two fragments: a F(ab')2 fragment and a pFc' (pepsin-cleaved Fc) fragment), Fab' (single chain of F(ab')2 fragment), A single chain variable fragment (scFv) (or a fusion protein of the variable regions of the heavy and light chains of an Ab linked together with a linker peptide, optionally a linker peptide of about 10 to about 25 amino acids in length), (scFv) 2, or two scFv or double scFv, or a single peptide chain having two variable heavy regions and two variable light regions to produce a tandem scFv, Minibodies (or fusion proteins of the variable regions of the Ab heavy and light chains linked together with an alkyl, optionally methyl or ethyl group) A diabody (or scFv, the scFv having a linker peptide that is too short for the two variable regions to fold together (optionally about 5 amino acids), thereby forcing the scFv to dimerize), a triabody or a tetrabody (or scFv, the scFv having a linker peptide that is too short for the two variable regions to fold together (optionally about 1 or 2 amino acids), thereby forcing the scFv to trimerize or tetramerize), Single domain antibodies (dAB) (or a single variable region of an Ab heavy chain or an Ab light chain), Multiple complementarity determining region (CDR) fragments, or Multispecific antibodies are formed from two or more antibody fragments.

3. The isolated or purified Ab or its Ag binding fragment, or monomer or dimer ABP according to any one of claims 1 or 2, wherein the heavy chain variable region (VH), if present, comprises: The following amino acid sequence: QSLEESGGRLVTPGTPLTLTCTVS GFSLSSYG VSWVRQAPGKGLEW IGIINSDHSTYYAKWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARSYG IGSIF WGPGTLVTVSS (SEQ ID NO:1), or SEQ ID NO: 1 having one or more amino acid substitutions, additions (insertions) or deletions, and the recombinant Ab or its Ag-binding fragment or monomeric or dimeric ABP retains its ability to specifically bind to the human ICOS protein or polypeptide.

4. The isolated or purified Ab or its Ag binding fragment, or monomeric or dimeric ABP according to claim 3, wherein: The one or more amino acid substitutions in the heavy chain variable region (VH) include one or more conservative amino acid substitutions.

5. The isolated or purified Ab or its Ag binding fragment or monomer or dimer ABP according to claim 4, wherein the one or more conservative amino acid substitutions comprise: Replace an aliphatic amino acid with another aliphatic amino acid; replace a serine with a threonine or vice versa; replace an acidic residue with another acidic residue; replace a residue bearing an amide group with another residue bearing an amide group; exchange a basic residue with another basic residue; or replace an aromatic residue with another aromatic residue.

6. An isolated or purified Ab or Ag binding fragment thereof, or a monomeric or dimeric ABP according to any one of claims 1 to 5 or according to any one of the preceding claims, wherein: The heavy chain variable region further comprises at least a portion of a heavy chain constant region.

7. The isolated or purified Ab or its Ag binding fragment, or monomer or dimer ABP according to claim 6, wherein the heavy chain constant region comprises the following amino acid sequence: GQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVTVTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVTSSSQPVTCNVAHPATNTKVDKTVAPSTCSKPTCPPPELLGGPSVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTA RPPLREQQFNSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPAPIEKTISKARGQPLEPKVYTMGPPREELSSRSVSLTCMINGFYPSDISVEWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSVPTSEWQRGDVFTCSVMHEALHNHYTQKSISRSPGK(SEQ ID NO:3).

8. The isolated or purified Ab or its Ag binding fragment, or monomer or dimer ABP according to any one of claims 1 to 7, wherein the heavy chain comprises a heavy chain variable region and a heavy chain constant region, and the heavy chain variable region and the heavy chain constant region comprise the following sequence: QSLEESGGRLVTPGTPLTLTCTVSGFSLSSYGVSWVRQAPGKGLEWIGIINSDHSTYYAKGRFTISKTSTTVDLKITSPTTEDTATYFCARSYGIGSIFWGPGTLVT VSSGQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVTVTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSVTSSSQPVTCNVAHPATNTKVDKTVAPSTCSKPTCPP PELLGGPSVFIFPPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPAPIEKTISKARG QPLEPKVYTMGPPREELSSRSVSLTCMINGFYPSDISVEWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSVPTSEWQRGDVFTCSVMHEALHNHYTQKSISRSPGK(SEQ ID NO:4).

9. An isolated or purified Ab or Ag-binding fragment thereof, or a monomeric or dimeric ABP according to any one of claims 1 to 7 or any of the preceding claims, wherein the heavy chain variable region comprises an amino-terminal signal sequence.

10. An isolated or purified Ab or Ag binding fragment thereof, or a monomeric or dimeric ABP according to claim 9 or any one of the preceding claims, wherein the heavy chain variable region amino acid terminal signal sequence comprises the sequence METGLRWLLLVAVLKGVQC (SEQ ID NO: 5).

11. An isolated or purified Ab or Ag binding fragment thereof, or a monomeric or dimeric ABP according to any one of claims 1 to 10 or according to any one of the preceding claims, The heavy chain variable region with a signal sequence comprises the following sequence METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLSSYGVSWVRQAPGKGLEWIGIINSDHSTYYAKWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARSYGIGSIFWGPGTLVTVSS(SEQ ID NO:6), or The heavy chain, comprising the variable and constant domains and having a signal sequence, comprises the following sequence: METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLSSYGVSWVRQAPGKGLEWIGIINSDHSTYYAKWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARS YGIGSIFWGPGTLVTVSSGQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVTVTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSVTSSSQPVTCNVAHPATNTKVDKTVAP STCSKPTCPPPELLGGPSVFIFPPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPAPIEKTI SKARGQPLEPKVYTMGPPREELSSRSVSLTCMINGFYPSDISVEWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSVPTSEWQRGDVFTCSVMHEALHNHYTQKSISRSPGK(SEQ ID NO:7).

12. An isolated or purified Ab or Ag binding fragment thereof, or a monomer or dimeric ABP according to any one of claims 1 to 11 or according to any one of the preceding claims, wherein the light chain variable region (VL), if present, comprises: (a) the following amino acid sequence: AAVLTQTPSPVSAAVGGTVSISCQSS KSVYNNNQ LSWFQQKPGQRP KLLIYEAFKLPSGVPSRFKGSGSGTQFTLTISDVQCDDAATYYCAAVYSD DSDNS FGGGTEVVVK (SEQ ID NO:2), or SEQ ID NO: 2 having one or more amino acid substitutions, additions (insertions) or deletions, and the recombinant Ab or its Ag-binding fragment or monomeric or dimeric ABP retains its ability to specifically bind to human ICOS protein or polypeptide.

13. The isolated or purified Ab or Ag-binding fragment thereof, or monomeric or dimeric ABP according to claim 12, wherein the one or more amino acid substitutions of the light chain variable region (VL) comprise one or more conservative amino acid substitutions.

14. The isolated or purified Ab or Ag binding fragment thereof, or monomer or dimer ABP according to claim 13, wherein the one or more conservative amino acid substitutions comprise: Conservative substitutions include: replacing an aliphatic amino acid with another aliphatic amino acid; replacing a serine with a threonine or vice versa; replacing an acidic residue with another acidic residue; replacing a residue bearing an amide group with another residue bearing an amide group; exchanging a basic residue with another basic residue; or replacing an aromatic residue with another aromatic residue.

15. An isolated or purified Ab or Ag binding fragment thereof, or a monomeric or dimeric ABP according to any one of claims 1 to 9 or according to any one of the preceding claims, wherein: The light chain variable region further comprises at least a portion of a light chain constant region.

16. The isolated or purified Ab or its Ag binding fragment, or monomer or dimer ABP according to claim 15, wherein the light chain constant region comprises the following amino acid sequence: GDPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTVTWEVDGTT QTTGIENSKTPQNSADCTYNLSSTLTLTSTQYNSHKEYTCKVTQGTTSVV QSFNRGDC (SEQ ID NO: 8).

17. The isolated or purified Ab or its Ag binding fragment, or monomer or dimer ABP according to claim 16, wherein the light chain comprises a variable region and a constant region, and the constant region comprises the following sequence: AAVLTQTPSPVSAAVGGTVSISCQSSKSVYNNNQLSWFQQKPGQRPKLLIYEAFKLPSGVPSRFKGSGSGTQFTLTISDVQCDDAATYYCAAVYSDDSDNSFGGGTEVV VKGDPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTVTWEVDGTTQTTGIENSKTPQNSADCTYNLSSTLTLTSTQYNSHKEYTCKVTQGTTSVVQSFNRGDC(SEQ IDNO:9).

18. An isolated or purified Ab or Ag binding fragment thereof, or a monomeric or dimeric ABP according to any one of claims 1 to 17 or according to any one of the preceding claims, wherein the light chain variable domain further comprises an amino-terminal signal sequence.

19. The isolated or purified Ab or Ag-binding fragment thereof, or monomeric or dimeric ABP according to claim 18, wherein the light chain variable domain amino-terminal signal sequence comprises the sequence MDTRAPTQLLGLLLLWLPGATF (SEQ ID NO: 10).

20. The isolated or purified Ab or Ag binding fragment thereof, or monomeric or dimeric ABP according to claim 19, wherein: The light chain variable domain with a signal sequence comprises or The light chain with a signal sequence comprises MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVSISCQSSKSVYNNNQLSWFQQKPGQRPKLLIYEAFKLPSGVPSRFKGSGSGTQFTLTISDVQCDDAATYYCAAVYSDDS DNSFGGGTEVVVKGDPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTVTWEVDGTTQTTGIENSKTPQNSADCTYNLSSTLTLTSTQYNSHKEYTCKVTQGTTSVVQSFNRGDC(SEQ ID NO:12).

21. An isolated or purified Ab or Ag-binding fragment thereof, or monomer or dimeric ABP according to any one of claims 1 to 20 or according to any of the preceding claims, wherein SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 11 and / or SEQ ID NO: 12 have two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen or fifteen conservative amino acid substitutions, and the recombinant Ab or Ag-binding fragment thereof, or monomer or dimeric ABP retains its ability to specifically bind to human ICOS protein or polypeptide.

22. An Ab or Ag binding fragment thereof, or a monomeric or dimeric ABP according to any one of claims 1 to 21 or according to any of the preceding claims, wherein the heavy chain constant region comprises an amino acid sequence from an IgG, IgM, IgA, IgD or IgE isotype.

23. An isolated or purified Ab or Ag binding fragment thereof, or a monomeric or dimeric ABP according to any one of claims 1 to 23 or according to any of the preceding claims, wherein the light chain constant region comprises an amino acid sequence from a kappa (κ) or lambda (λ) isotype.

24. An Ab or Ag binding fragment thereof, or a monomer or dimeric ABP according to any one of claims 1 to 23 or according to the isolated or purified Ab of any one of the preceding claims, wherein at least a portion of the heavy chain constant region, at least a portion of the light chain constant region, or at least a portion of the heavy chain constant region and the light chain constant region is or comprises an amino acid sequence of human, rabbit, mouse or rat origin or comprises a constant region amino acid sequence derived from human, rabbit, mouse or rat.

25. An isolated or purified Ab or Ag-binding fragment thereof, or a monomeric or dimeric ABP according to any one of claims 1 to 24 or according to any of the preceding claims, wherein at least a portion of the heavy chain constant region, at least a portion of the light chain constant region, or at least a portion of the heavy chain constant region and the light chain constant region is or comprises a synthetic amino acid sequence.

26. An isolated or purified Ab or Ag-binding fragment thereof, or monomer or dimeric ABP according to any one of claims 1 to 25 or according to any of the preceding claims, wherein the recombinant Ab, its Ag-binding fragment, or monomer or dimeric ABP, or the heavy chain constant region, or the light chain constant region, or the heavy chain constant region and the light chain constant region further comprise or are bound to a heterologous protein, peptide, or compound or composition.

27. The isolated or purified Ab or Ag-binding fragment thereof, or monomeric or dimeric ABP according to claim 26, wherein the heterologous protein or peptide, or the compound or composition comprises a detectable protein, a detectable agent or a binding moiety.

28. The isolated or purified Ab or Ag-binding fragment thereof, or monomeric or dimeric ABP according to claim 26, wherein the heterologous protein or peptide comprises a carrier protein.

29. The isolated or purified Ab or its Ag-binding fragment, or monomer or dimeric ABP according to any one of claims 26 to 28, wherein the heterologous protein, peptide or the compound or composition is covalently conjugated to the recombinant antibody (Ab) or its Ag-binding fragment, or monomer or dimeric ABP.

30. An isolated or purified Ab or Ag binding fragment thereof, or monomer or dimeric ABP according to any one of claims 26 to 29, wherein the detectable agent or binding moiety comprises biotin, a fluorescent or chemiluminescent label, a fluorophore, perylene, fluorenyl, coumarin, 7-methoxycoumarin (Mca), 4-(dimethylaminoazo)benzene-4-carboxylic acid (dabcyl), Tamra, boron dipyrromethene (BODIPY) or a derivative thereof, a dye, a radioactive isotope, quantum dots or photoluminescent aqueous nanocrystals, a hapten or an antibody binding epitope or domain.

31. The isolated or purified Ab or Ag-binding fragment thereof, or monomeric or dimeric ABP according to claim 31, wherein the dye is or comprises rhodamine, [2-(4-nitro-2,1,3-benzoxadiazol-7-yl)aminoethyl]trimethylammonium (NBD), Nile Red or Nile Blue, or is a fluorescent dye comprising sulfoindocyanine.

32. The isolated or purified Ab or Ag-binding fragment thereof, or monomeric or dimeric ABP according to claim 31, wherein the fluorophore is or comprises a dansyl, fluorescein, carboxyfluorescein (FAM) or 6-FAM moiety.

33. The isolated or purified Ab or Ag-binding fragment thereof, or monomeric or dimeric ABP according to claim 31, wherein the dye is or comprises a cyanine dye, Cy3 or Cy5.

34. The isolated or purified Ab or Ag-binding fragment thereof, or monomeric or dimeric ABP according to claim 31, wherein the hapten is or comprises a biotin, theophylline, digitoxin, carborane, fluorescein or bromodeoxyuridine moiety.

35. An isolated or purified Ab or Ag-binding fragment thereof, or monomer or dimeric ABP according to any one of claims 1 to 35 or according to any of the preceding claims, wherein the Ab or Ag-binding fragment thereof, or monomer or dimeric ABP is a recombinant Ab or Ag-binding fragment thereof, or monomer or dimeric ABP, or comprises a peptide or polypeptide made by recombinant technology.

36. A chimeric or recombinant nucleic acid comprising: a nucleic acid sequence encoding the Ab or Ag binding fragment thereof, or a monomeric or dimeric ABP according to any one of claims 1 to 35.

37. The chimeric or recombinant nucleic acid of claim 36, wherein the chimeric or recombinant nucleic acid further comprises and is operably linked to a transcriptional regulatory element.

38. The chimeric or recombinant nucleic acid of claim 37, wherein the transcriptional regulatory element comprises a promoter.

39. The chimeric or recombinant nucleic acid of claim 38, wherein the promoter is an inducible promoter or a constitutive promoter.

40. The chimeric or recombinant nucleic acid of any one of claims 36 to 39, wherein the chimeric or recombinant nucleic acid further comprises a sequence encoding an amino-terminal signal peptide.

41. The chimeric or recombinant nucleic acid of claim 40, wherein the amino terminal signal peptide comprises the following amino acid sequence: METGLRWLLLVAVLKGVQC (SEQ ID NO: 5); or MDTRAPTQLLGLLLLWLPGATF (SEQ ID NO: 10).

42. An expression cassette, vector, recombinant virus, artificial chromosome, cosmid or plasmid, comprising or containing the chimeric or recombinant nucleic acid according to any one of claims 36 to 41.

43. A cell comprising or containing: a chimeric or recombinant antibody or dimeric antigen binding protein according to any one of claims 1 to 35, a chimeric or recombinant nucleic acid according to any one of claims 36 to 41, or an expression cassette, vector, recombinant virus, artificial chromosome, cosmid or plasmid according to claim 42.

44. The cell of claim 43, wherein the cell is a bacterial, fungal, mammalian, yeast, insect or plant cell.

45. The cell of claim 44, wherein the mammalian cell is a human cell.

46. ​​A method for detecting the presence of human ICOS (inducible T-cell co-stimulatory) protein or human CD278 protein or polypeptide in or on a cell, tissue, organ or a portion of any of the foregoing, the method comprising: (a) contacting the cell, tissue or organ, or a portion of any of the foregoing, with at least one Ab or Ag-binding fragment thereof, or monomeric or dimeric ABP according to any one of claims 1 to 35 or according to any of the preceding claims, and (b) detecting specific binding of said at least one Ab or Ag-binding fragment thereof, or monomeric or dimeric ABP to a BCMA polypeptide in or on said cell, tissue or organ, or a portion of any of the foregoing, Thereby detecting the presence of the human BCMA protein in or on the cell, tissue or organ or a part of any of the foregoing.

47. The method of claim 46, wherein the at least one Ab or Ag-binding fragment thereof or monomer or dimeric ABP comprises a variable heavy chain (VH) having an amino acid sequence comprising SEQ ID NO: 1 and a variable light chain (VL) having an amino acid sequence comprising SEQ ID NO:

2.

48. The method of claim 47, wherein the at least one Ab or Ag-binding fragment thereof or monomeric or dimeric ABP comprises a heavy chain having an amino acid sequence comprising SEQ ID NO:4 and a light chain having an amino acid sequence comprising SEQ ID NO:

9.

49. The method of any one of claims 46 to 48, wherein the method comprises contacting the cell, tissue or organ, or a portion of any of the foregoing, with two Abs or Ag-binding fragments thereof, or monomeric or dimeric ABPs, or a mixture of two Abs or Ag-binding fragments thereof, or monomeric or dimeric ABPs.

50. The method of any one of claims 46 to 49, wherein the contacting comprises using an immunohistochemistry (IHC) assay.

51. The method according to any one of claims 46 to 50, further comprising contacting the Ab or its Ag-binding fragment, or monomer or dimeric ABP that specifically binds to human ICOS protein or human CD278 protein or polypeptide with a detectable agent to indicate or signal the specific binding of the Ab or its Ag-binding fragment, or monomer or dimeric ABP to human ICOS protein or human CD278 protein or polypeptide.

52. The method of claim 51, wherein the detectable agent specifically binds to the Ab or Ag-binding fragment thereof, or monomeric or dimeric ABP.

53. The method of any one of claims 46 to 52, wherein the cell, tissue, organ, or a portion of any of the foregoing is or comprises: Early B cells, progenitor B cells, pre-B lymphocytes, mature B lymphocytes, follicle center cells, or cells in the germinal center of tonsils, organs, or lymph nodes, bone marrow stem cells, bone marrow hematopoietic cells, T lymphocytes, parafollicular T lymphocytes, subpopulations of parafollicular T lymphocytes, hepatobiliary tubule cells, glomerular cells, proximal tubule cells, mammary myoepithelial cells, stromal cells surrounding or associated with infiltrating tumor cells, kidney cells, cerebellar cells, prostate cells, pancreatic cells, bone marrow cells, or epithelial cells.

54. The method of claim 53, wherein the epithelial cell is a brain, lung, intestinal, kidney, breast or placental epithelial cell.

55. The method of any one of claims 46 to 54, wherein the organ is liver, prostate, brain, pancreas, bladder, colon, esophagus, kidney, skin or lung.

56. The method of any one of claims 46 to 55, wherein the cell, tissue, organ, or a portion of any of the foregoing is or comprises a cancer or tumor cell, or a malignant tumor cell or a carcinoid tumor cell.

57. The method of claim 52, wherein the cancer or tumor cell is or comprises or is derived from: a malignant tumor cell (optionally a squamous cell carcinoma cell), a breast cancer cell, a colon cancer or colorectal cancer cell, a melanoma cell (optionally a malignant melanoma cell) or a multiple myeloma cell, a plasmacytoma cell, a lymphoma cell, a bladder or urothelial cancer cell, a cervical cancer cell, an ovarian cancer cell, an esophageal cancer or an esophageal squamous cell carcinoma cell; a malignant pleural mesothelioma cell, a prostate cancer cell, a cell from a microsatellite instability-high / mismatch repair-deficient tumor, a human papillomavirus-positive or Epstein-Barr-positive tumor cell, a hepatocellular carcinoma or a liver cancer cell, a lung cancer cell (optionally a non-small cell lung cancer (NSCLC)), a gastric cancer cell, a renal cancer cell, a pancreatic cancer cell, a breast cancer cell (optionally a triple-negative breast cancer cell), a lymphoma cell or a leukemia cell, or a mycosis fungoides cell.

58. The method of claim 49, wherein the lymphoma or leukemia cells are or are derived from: B-cell lymphoma, acute lymphoblastic leukemia (ALL) cells, angioimmunoblastic T-cell lymphoma cells, cutaneous T-cell lymphoma, chronic myeloid leukemia in blast crisis, diffuse large B-cell lymphoma cells, hairy cell leukemia cells, follicular lymphoma cells, Burkitt's lymphoma, diffuse large B-cell lymphoma, or mantle cell lymphoma.

59. The method of claim 58, wherein the B cell lymphoma is Hodgkin lymphoma or non-Hodgkin lymphoma.

60. The method of claim 59, wherein the non-Hodgkin lymphoma is follicular lymphoma, diffuse large B-cell lymphoma, marginal zone B-cell lymphoma, small lymphocytic lymphoma (SLL) or chronic lymphocytic leukemia (CLL), Burkitt's lymphoma or mantle cell lymphoma (MCL).

61. The method of claim 56, wherein the cancer cells are head and neck squamous cell carcinoma (HNSCC) or basal cell carcinoma (BCC) cells.

62. A method for detecting or diagnosing cancer or a tumor, The method comprises detecting the expression or presence of human ICOS (inducible T cell co-stimulatory) protein, or human CD278 protein or polypeptide in or on a cell, tissue or organ sample using the method according to any one of claims 46 to 61, Wherein the specific binding of the Ab or its Ag binding fragment or monomer or dimer ABP to the human ICOS (inducible T cell co-stimulatory) protein or human CD278 protein or polypeptide in or on the cell, tissue or organ or a part of any of the foregoing is detected to detect or diagnose or assist in the detection or diagnosis of the cancer or the tumor.

63. The method of claim 62, wherein the cancer or the tumor is or comprises or is derived from: a malignant tumor cell (optionally a squamous cell carcinoma cell), or a breast cancer cell, a colon cancer cell or a colorectal cancer cell, a melanoma cell (optionally a malignant melanoma cell) or a multiple myeloma cell, a plasmacytoma cell, a lymphoma cell, a bladder or urothelial cancer cell, a cervical cancer cell, an ovarian cancer cell, an esophageal cancer or an esophageal squamous cell carcinoma cell; a malignant pleural mesothelioma cell, a prostate cancer cell, a cell from a microsatellite instability-high / mismatch repair-deficient tumor, a human papillomavirus-positive or Epstein-Barr-positive tumor cell, a hepatocellular carcinoma or a liver cancer cell, a lung cancer cell (optionally a non-small cell lung cancer (NSCLC)), a gastric cancer cell, a renal cancer cell, a pancreatic cancer cell, a breast cancer cell (optionally a triple-negative breast cancer cell), a lymphoma cell or a leukemia cell, or a mycosis fungoides cell.

64. The method of claim 63, wherein the lymphoma or leukemia cells are or are derived from: B-cell lymphoma, acute lymphoblastic leukemia (ALL) cells, angioimmunoblastic T-cell lymphoma cells, cutaneous T-cell lymphoma, chronic myeloid leukemia in blast crisis, diffuse large B-cell lymphoma cells, hairy cell leukemia cells, follicular lymphoma cells, Burkitt's lymphoma, diffuse large B-cell lymphoma, or mantle cell lymphoma.

65. The method of claim 64, wherein the B cell lymphoma is Hodgkin lymphoma or non-Hodgkin lymphoma.

66. The method of claim 65, wherein the non-Hodgkin lymphoma is follicular lymphoma, diffuse large B-cell lymphoma, marginal zone B-cell lymphoma, small lymphocytic lymphoma (SLL) or chronic lymphocytic leukemia (CLL), Burkitt's lymphoma or mantle cell lymphoma (MCL).

67. The method of claim 63, wherein the cancer cells are head and neck squamous cell carcinoma (HNSCC) or basal cell carcinoma (BCC) cells.

68. The method of any one of claims 62 to 67, wherein the cell, tissue or organ sample is from an individual in need or an individual at increased risk of developing a cancer or tumor or having a family history of said cancer or tumor.

69. The method of any one of claims 62 to 68, wherein the detecting comprises performing an immunohistochemistry (IHC) assay.

70. The method of any one of claims 62 to 69, wherein at least two Abs or Ag-binding fragments thereof, or monomeric or dimeric ABPs are contacted with the cell, tissue or organ sample.

71. The method of claim 70, wherein at least one of the two Abs or Ag-binding fragments thereof or monomeric or dimeric ABPs comprises a variable heavy chain (VH) having an amino acid sequence comprising SEQ ID NO: 1 and a variable light chain (VL) having an amino acid sequence comprising SEQ ID NO:

2.

72. The method of claim 71, wherein the at least one Ab or Ag-binding fragment thereof or monomeric or dimeric ABP comprises a heavy chain having an amino acid sequence comprising SEQ ID NO:4 and a light chain having an amino acid sequence comprising SEQ ID NO:

9.

73. A method for treating, improving or preventing cancer or tumors, the method comprising first detecting or diagnosing the cancer or tumor using the method according to any one of claims 62 to 72, and then treating the individual in need to treat, improve or prevent the cancer or tumor.

74. The method of claim 73, wherein the cancer or the tumor is or comprises or is derived from: a malignant tumor cell (optionally a squamous cell carcinoma cell), a breast cancer cell, a colon cancer cell or a colorectal cancer cell, a melanoma cell (optionally a malignant melanoma cell) or a multiple myeloma cell, a plasmacytoma cell, a lymphoma cell, a bladder or urothelial cancer cell, a cervical cancer cell, an ovarian cancer cell, an esophageal cancer or an esophageal squamous cell carcinoma cell; a malignant pleural mesothelioma cell, a prostate cancer cell, a cell from a microsatellite instability-high / mismatch repair-deficient tumor, a human papillomavirus-positive or Epstein-Barr-positive tumor cell, a hepatocellular carcinoma or a liver cancer cell, a lung cancer cell (optionally a non-small cell lung cancer (NSCLC)), a gastric cancer cell, a renal cancer cell, a pancreatic cancer cell, a breast cancer cell (optionally a triple-negative breast cancer cell), a lymphoma cell or a leukemia cell, or a mycosis fungoides cell.

75. The method of claim 74, wherein the lymphoma or leukemia cells are or are derived from: B-cell lymphoma, acute lymphoblastic leukemia (ALL) cells, angioimmunoblastic T-cell lymphoma cells, cutaneous T-cell lymphoma, chronic myeloid leukemia in blast crisis, diffuse large B-cell lymphoma cells, hairy cell leukemia cells, follicular lymphoma cells, Burkitt's lymphoma, diffuse large B-cell lymphoma, or mantle cell lymphoma.

76. The method of claim 75, wherein the B cell lymphoma is Hodgkin lymphoma or non-Hodgkin lymphoma.

77. The method of claim 76, wherein the non-Hodgkin lymphoma is follicular lymphoma, diffuse large B-cell lymphoma, marginal zone B-cell lymphoma, small lymphocytic lymphoma (SLL), or chronic lymphocytic leukemia, Burkitt's lymphoma, or mantle cell lymphoma (MCL).

78. The method of claim 74, wherein the cancer cells are head and neck squamous cell carcinoma (HNSCC) or basal cell carcinoma (BCC) cells.

79. The method of any one of claims 73 to 78, wherein the cell, tissue or organ sample is from an individual in need, and optionally, the individual in need is an individual at higher risk of developing a cancer or tumor or has a family history of the cancer or tumor.

80. Use of at least one recombinant antibody (Ab) or its antigen (Ag) binding fragment, or monomeric or dimeric antigen binding protein (ABP) according to any one of claims 1 to 35 or encoded by the nucleic acid according to any one of claims 36 to 41 for detecting or diagnosing cancer, or treating, ameliorating or preventing cancer.

81. The use of claim 80, wherein the cancer or the tumor is or comprises or is derived from: a malignant tumor cell (optionally a squamous cell carcinoma cell), a breast cancer cell, a colon cancer cell or a colorectal cancer cell, a melanoma cell (optionally a malignant melanoma cell) or a multiple myeloma cell, a plasmacytoma cell, a lymphoma cell, a bladder or urothelial cancer cell, a cervical cancer cell, an ovarian cancer cell, an esophageal cancer or an esophageal squamous cell carcinoma cell; a malignant pleural mesothelioma cell, a prostate cancer cell, a cell from a microsatellite instability-high / mismatch repair-deficient tumor, a human papillomavirus-positive or Epstein-Barr-positive tumor cell, a hepatocellular carcinoma or a liver cancer cell, a lung cancer cell (optionally a non-small cell lung cancer (NSCLC)), a gastric cancer cell, a renal cancer cell, a pancreatic cancer cell, a breast cancer cell (optionally a triple-negative breast cancer cell), a lymphoma cell or a leukemia cell, or a mycosis fungoides cell.

82. The method of claim 81, wherein the lymphoma or leukemia cells are or are derived from: B-cell lymphoma, acute lymphoblastic leukemia (ALL) cells, angioimmunoblastic T-cell lymphoma cells, cutaneous T-cell lymphoma, chronic myeloid leukemia in blast crisis, diffuse large B-cell lymphoma cells, hairy cell leukemia cells, follicular lymphoma cells, Burkitt's lymphoma, diffuse large B-cell lymphoma, or mantle cell lymphoma.

83. The method of claim 82, wherein the B cell lymphoma is Hodgkin lymphoma or non-Hodgkin lymphoma.

84. The method of claim 83, wherein the non-Hodgkin lymphoma is follicular lymphoma, diffuse large B-cell lymphoma, marginal zone B-cell lymphoma, small lymphocytic lymphoma (SLL), or chronic lymphocytic leukemia, Burkitt's lymphoma, or mantle cell lymphoma (MCL).

85. The method of claim 81, wherein the cancer cell is a head and neck squamous cell carcinoma (HNSCC) or a basal cell carcinoma (BCC) cell.

86. The use according to any one of claims 80 to 85, wherein the detecting comprises performing an immunohistochemistry (IHC) assay.

87. A recombinant antibody (Ab) or its antigen (Ag) binding fragment, or a monomer or dimeric antigen binding protein (ABP) according to any one of claims 1 to 35 or according to any one of the preceding claims, wherein the recombinant antibody (Ab) or its antigen (Ag) binding fragment, or a monomer or dimeric antigen binding protein (ABP) is used to detect or diagnose cancer, or to treat, improve or prevent cancer.

88. The recombinant antibody (Ab) or antigen (Ag) binding fragment thereof, or monomeric or dimeric antigen binding protein (ABP) according to claim 80, wherein the cancer or the tumor is or comprises or is derived from: a malignant tumor cell (optionally a squamous cell carcinoma cell), a breast cancer cell, a colon cancer cell or a colorectal cancer cell, a melanoma cell (optionally a malignant melanoma cell) or a multiple myeloma cell, a plasmacytoma cell, a lymphoma cell, a bladder or urothelial carcinoma cell, a cervical cancer cell, an ovarian cancer cell, an esophageal cancer or an esophageal squamous cell carcinoma cell; a malignant pleural mesothelioma cell, a prostate cancer cell, a cell from a microsatellite instability-high / mismatch repair-deficient tumor, a human papillomavirus-positive or Epstein-Barr-positive tumor cell, a hepatocellular carcinoma or a liver cancer cell, a lung cancer cell (optionally a non-small cell lung cancer (NSCLC)), a gastric cancer cell, a renal cancer cell, a pancreatic cancer cell, a breast cancer cell (optionally a triple-negative breast cancer cell), a lymphoma cell or a leukemia cell, or a mycosis fungoides cell.

89. The method of claim 88, wherein the lymphoma or leukemia cells are or are derived from: B-cell lymphoma, acute lymphoblastic leukemia (ALL) cells, angioimmunoblastic T-cell lymphoma cells, cutaneous T-cell lymphoma, chronic myeloid leukemia in blast crisis, diffuse large B-cell lymphoma cells, hairy cell leukemia cells, follicular lymphoma cells, Burkitt's lymphoma, diffuse large B-cell lymphoma, or mantle cell lymphoma.

90. The method of claim 89, wherein the B cell lymphoma is Hodgkin lymphoma or non-Hodgkin lymphoma.

91. The method of claim 90, wherein the non-Hodgkin lymphoma is follicular lymphoma, diffuse large B-cell lymphoma, marginal zone B-cell lymphoma, small lymphocytic lymphoma (SLL), or chronic lymphocytic leukemia, Burkitt's lymphoma, or mantle cell lymphoma (MCL).

92. The method of claim 88, wherein the cancer cells are head and neck squamous cell carcinoma (HNSCC) or basal cell carcinoma (BCC) cells.

93. according to any one of claims 87 to 92 recombinant antibodies (Ab) or their antigen (Ag) binding fragments, or monomeric or dimeric antigen binding proteins (ABP), wherein said detection or diagnosis comprises performing an immunohistochemistry (IHC) assay.

94. A kit comprising: a chimeric or recombinant antibody according to any one of claims 1 to 35 or according to any one of the preceding claims; a chimeric or recombinant nucleic acid according to any one of claims 36 to 41; or an expression cassette, vector, recombinant virus, artificial chromosome, cosmid or plasmid according to claim 42; or a cell according to any one of claims 43 to 45.

95. The kit of claim 94, wherein the kit comprises components required for an immunohistochemistry (IHC) assay, and / or comprises instructions for practicing the method according to any one of the preceding claims.

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