# US 9,518,120 B2 · Anti-CXCR4 antibody and its use for the detection and diagnosis of cancer > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,518,120 B2 - Document ID: US9518120B2 - Status: Granted - Kind: B2 - Application number: 14/235,274 - Filed: 2012-07-30 - Published: 2014-06-19 - Granted: 2016-12-13 - Earliest claimed: 2011-07-29 - Assignees: Pierrre Fabre Medicament - Applicants: Christine Klinguer-Hamour; Alexandra Jouhanneaud; Marie-Claire Janin-Bussat - Inventors: Christine Klinguer-Hamour; Alexandra Jouhanneaud; Marie-Claire Janin-Bussat - Web page: https://patentgrep.com/patent/US9518120B2 ## CPC - G01N 33/57575: {involving oncogenic proteins} - A61P 35/00: Antineoplastic agents - C07K 16/2866: {against receptors for cytokines, lymphokines, interferons} - C07K 16/2896: {against molecules with a "CD"-designation, not provided for elsewhere} - G01N 33/575: for cancer - C07K 2317/30: characterized by aspects of specificity or valency - C07K 2317/51: Complete heavy chain or Fd fragment, i.e. VH + CH1 - C07K 2317/515: Complete light chain, i.e. VL + CL - C07K 2317/565: Complementarity determining region [CDR] - G01N 2333/7158: for chemokines - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract The present invention provides a novel, isolated anti-CXCR4 antibody for use in the diagnosis of cancer. In particular, the antibody of the invention recognizes monomeric and homodimeric CXCR4, but not heterodimeric CXCR4. ## Claims ### Claim 1 An anti-CXCR4 antibody, or a CXCR4-binding fragment thereof, comprising i) a heavy chain comprising the following three CDRs, respectively CDR-H1 having the sequence SEQ ID No. 1, CDR-H2 having the sequence SEQ ID No. 2, and CDR-H3 having the sequence SEQ ID No. 3; and ii) a light chain comprising the following three CDRs, respectively CDR-L1 having the sequence SEQ ID No. 4, CDR-L2 having the sequence SEQ ID No. 5, and CDR-L3 having the sequence SEQ ID No. 6. ### Claim 2 (depends on 1) The antibody of claim 1, or a CXCR4-binding fragment thereof, wherein the said antibody is selected among: a. an antibody with a heavy chain comprising the following three CDRs, respectively CDR-H1 having the sequence SEQ ID No. 1, CDR-H2 having the sequence SEQ ID No. 2 and CDR-H3 having the sequence SEQ ID No. 3; and a light-chain variable domain comprising the sequence SEQ ID No. 8; b. an antibody with a heavy chain variable domain comprising the sequence SEQ ID No. 7; and a light chain comprising the following three CDRs, respectively CDR-L1 having the sequence SEQ ID No. 4, CDR-L2 having the sequence SEQ ID No. 5 and CDR-L3 having the sequence SEQ ID No. 6; or c. an antibody with a heavy chain variable domain comprising the sequence SEQ ID No. 7; and a light-chain variable domain comprising the sequence SEQ ID No. 8. ### Claim 3 (depends on 1) The antibody according to claim 1, or a CXCR4-binding fragment thereof, wherein said antibody is a murine antibody. ### Claim 4 A murine hybridoma capable of secreting an antibody or a CXCR4-binding fragment thereof, said murine hybridoma being deposited at the CNCM, Institut Pasteur, Paris, France on Jun. 25, 2008 under the number I-4018. ### Claim 5 (depends on 1, 4) A method for detecting the presence of a tumor expressing monomeric/homodimeric CXCR4 in a subject, wherein said process comprises the steps of: (a) contacting a tumor sample from the said subject with an antibody, or a CXCR4-binding fragment thereof, according to claim 1 or produced by a hybridoma according to claim 4; (b) detecting the binding of the said antibody, or a CXCR4-binding fragment thereof, with the tumor sample, wherein said binding indicates the presence of a tumor expressing monomeric/homodimeric CXCR4 in said subject. ### Claim 6 (depends on 1, 4) A method for determining the percentage of cells expressing CXCR4 as monomer and/or homodimer in a tumor from a subject, said method comprising the steps of: (a) contacting a tumor sample from the subject with an antibody, or a CXCR4-binding fragment thereof, according to claim 1 or produced by a hybridoma according to claim 4; and (b) quantifying the percentage of cells expressing CXCR4 as monomer and/or homodimer in the sample. ### Claim 7 (depends on 1, 4) A method for determining the expression level of monomeric/homodimeric CXCR4 as monomer and/or homodimer in a tumor from a subject, said method comprising the steps of: (a) contacting a tumor sample from the subject with an antibody or a CXCR4-binding fragment thereof, according to claim 1 or produced by a hybridoma according to claim 4; and (b) quantifying the level of binding of the said antibody, or a CXCR4-binding fragment thereof, to monomeric/homodimeric CXCR4 in the tumor sample. ### Claim 8 (depends on 7) The method of claim 7, wherein the level of binding of the said antibody, or a CXCR4-binding fragment thereof, to monomeric/homodimeric CXCR4 is measured by immunohistochemistry (IHC) or FACS, preferably by IHC. ### Claim 9 (depends on 1, 4) A method for determining the scoring of a tumor from a subject, said method comprising: (a) contacting a tumor sample from the subject with an antibody, or or a CXCR4-binding fragment thereof, according to claim 1 or produced by a hybridoma according to claim 4; (b) quantifying the level of binding of said antibody, or or a CXCR4-binding fragment thereof, to monomeric/homodimeric CXCR4 as monomer and/or homodimer in the said tumor sample; and (c) scoring the tumor by comparing the quantified level of binding of the said antibody, or a CXCR4-binding fragment thereof, from the subject to an appropriate scale. ### Claim 10 (depends on 9) The method of claim 9, wherein the said appropriate scale is based on two parameters which are the intensity of the staining and the percentage of positive cells. ### Claim 11 (depends on 9) The method of claim 9, wherein the said appropriate scale is a scale of 0 to 8 wherein “no reactivity” is scored 0, and a strong reactivity in a proportion of “67-100% proportion reactive” is scored 8. ### Claim 12 (depends on 9) A method for determining the status of a tumor from a subject, said method comprising the steps of: (a) scoring a tumor from a subject according to claim 9 and b) determining that the status of the tumor is [monomeric/homodimeric CXCR4(+)] with a score of 3 to 8; or (c) determining that the status of the tumor is [monomeric/homodimeric CXCR4(−)] with a score of 0 to 2. ### Claim 13 (depends on 9) The method of claim 9, wherein the said appropriate scale is a scale of 0 to 3+ wherein no membranous reactivity of tumor cells is scored 0, and strong complete reactivity in more than 10% of tumor cells is scored 3+. ### Claim 14 (depends on 10) A method for determining the status of a tumor from a subject, said method comprising the steps of: (a) scoring a tumor from a subject according to claim 10; and (b) determining that the status of the tumor is [monomeric/homodimeric CXCR4(+)] with a score of 2+ or 3+; or (c) determining that the status of the tumor is [monomeric/homodimeric CXCR4(−)] with a score of 0 or 1+. ### Claim 15 (depends on 1, 4) A kit comprising at least an antibody, or a CXCR4-binding fragment thereof, according to claim 1 or produced by a hybridoma according to claim 4. ### Claim 16 (depends on 15) The kit of claim 15, characterized in that said antibody, or a CXCR4-binding fragment thereof, is labeled. ### Claim 17 (depends on 15) The kit of claim 15, further comprising positive and negative control samples for the scoring of monomeric/homodimeric CXCR4 expression level.