# US 9,040,018 B2 · Monoclonal antibodies against alpha-actinin-4 antigens, and uses therefor > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,040,018 B2 - Document ID: US9040018B2 - Status: Granted - Kind: B2 - Application number: 13/976,811 - Filed: 2011-12-23 - Published: 2014-03-06 - Granted: 2015-05-26 - Earliest claimed: 2010-12-29 - Applicants: Özge Alper - Inventors: Özge Alper - Web page: https://patentgrep.com/patent/US9040018B2 ## CPC - C07K 16/18: against material from animals or humans - G01N 33/57515: of the breast - G01N 33/5758: {involving compounds serving as markers for tumours, cancers or neoplasias, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides or metabolites} - G01N 2333/4712: Muscle proteins, e.g. myosin, actin, protein - 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) - 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) ## Abstract Monoclonal antibodies (MoAbs or mAbs) specific for ALPHA-ACTININ-4 antigens, hybridoma lines that secrete these ALPHA-ACTININ-4 mAbs, and the use of such mAbs to detect ALPHA-ACTININ-4 antigens, particularly those expressed by cancer cells are disclosed. Chimeric and humanized antibodies based upon these anti-ALPHA-ACTININ-4 mAbs, processes for producing monoclonal, chimeric, and humanized antibodies using recombinant DNA technology, and their therapeutic uses, particularly in the treatment of cancer are also disclosed. Methods and kits for the immunodetection and immunotherapy of cells for samples which express ALPHA-ACTININ-4 antigens are additionally disclosed. ## Claims ### Claim 1 An isolated antibody or antibody fragment that binds ALPHA-ACTININ-4, comprising a heavy chain variable domain comprising three complementary determining regions (CDRs) comprising the amino acid sequences of SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4, and a light chain variable domain comprising three CDRs comprising the amino acid sequences of SEQ ID NO: 6, SEQ ID NO: 7; and SEQ ID NO: 8. ### Claim 2 (depends on 1) The antibody or antibody fragment according to claim 1 wherein said ALPHA-ACTININ-4 is a soluble protein having a molecular weight of about 95 to 104 kilodaltons as measured by gradient polyacrylamide gel electrophoresis. ### Claim 3 (depends on 1) The antibody or antibody fragment according to claim 1, wherein the antibody or antibody fragment is immobilized on a solid phase. ### Claim 4 (depends on 1) The antibody or antibody fragment according to claim 1, wherein the antibody or antibody fragment is detectably labeled. ### Claim 5 (depends on 1) The antibody or antibody fragment according to claim 1, wherein the antibody or antibody fragment is conjugated to a cytokine, cytotoxin, radionuclide, drug, immunomodulator, therapeutic enzyme, anti-proliferative agent, cytotoxic radionuclide, cytotoxic drug, or cytotoxic protein. ### Claim 6 (depends on 1) The antibody or antibody fragment of claim 1 specific for a nuclear or soluble form of ALPHA-ACTININ-4. ### Claim 7 (depends on 1) The antibody or antibody fragment of claim 1, wherein the antibody or antibody fragment is humanized. ### Claim 8 (depends on 1) A pharmaceutical composition comprising the antibody or antibody fragment according to claim 1 in combination with a pharmaceutically acceptable carrier. ### Claim 9 (depends on 1) A recombinant nucleic acid encoding the antibody or antibody fragment of claim 1. ### Claim 10 (depends on 9) An isolated vector comprising the nucleic acid of claim 9. ### Claim 11 (depends on 10) An isolated host cell comprising the vector of claim 10. ### Claim 12 (depends on 11) An isolated antibody or antibody fragment produced by a method comprising culturing the host cell of claim 11, expressing the antibody or antibody fragment, and recovering the antibody or antibody fragment expressed by the host cell. ### Claim 13 (depends on 1) An immunoassay for detecting ALPHA-ACTININ-4 in a biological sample, comprising contacting said biological sample with the antibody or antibody fragment of claim 1, and the presence or absence of ALPHA-ACTININ-4 is determined by qualitatively or quantitatively detecting the presence or absence of an antibody bound to ALPHA-ACTININ-4. ### Claim 14 (depends on 13) The immunoassay of claim 13, wherein the biological sample comprises blood, tissue, or cells. ### Claim 15 (depends on 14) The immunoassay of claim 14, wherein the blood sample is whole blood, plasma, or serum. ### Claim 16 (depends on 14) The immunoassay of claim 14, wherein the tissue or cell is from breast, ovary, head, neck, or brain. ### Claim 17 (depends on 13) The immunoassay of claim 13, wherein the sample comprises circulating tumor cells. ### Claim 18 (depends on 1) A method for diagnosing breast cancer comprising: (a) obtaining a specimen from a human patient; (b) contacting the specimen with the antibody or antibody fragment of claim 1; (c) quantitatively determining whether the antibody or antibody fragment specifically binds to said specimen; (d) comparing the level of specific binding to a control, wherein binding at a level that is lower than the control indicates the presence of breast cancer in said specimen. ### Claim 19 (depends on 1, 4) A kit comprising: i) the isolated antibody or antibody fragment of claim 1 and a secondary antibody that binds to the antibody or antibody fragment of claim 1, wherein the secondary antibody is conjugated to a detectable label; or ii) the isolated antibody or antibody fragment of claim 4; and instructions for use. ### Claim 20 (depends on 1) An immunohistochemical method of detecting breast cancer in a cell, tissue, or blood sample collected from a human subject comprising the steps of: a) obtaining a tissue, cell, or blood sample from a human subject; b) contacting said sample with the antibody or antibody fragment of claim 1; c) quantitatively determining whether the antibody specifically binds to said sample; d) comparing the level of specific binding in the sample to a control, wherein binding at a level that is lower than in the control indicates the presence of breast cancer. ### Claim 21 (depends on 1) A method of treating cancer with an anti-ALPHA-ACTININ-4 antibody or antibody fragment comprising obtaining a tissue, cell, or blood sample from a human subject; applying the antibody or antibody fragment of claim 1 to the sample; quantitatively determining the level of specific binding of the antibody as compared to a control; and providing an anti-ALPHA-ACTININ-4 antibody treatment to said patient if the antibody or antibody fragment specifically binds at a level that is less than the level of binding in the control. ### Claim 22 (depends on 1) A prognostic method for determining the severity of disease in a patient diagnosed with or suspected of having breast cancer comprising: a) obtaining a cell or tissue sample from a human subject diagnosed with or suspected of having breast cancer; b) contacting the sample with the antibody or antibody fragment of claim 1; c) detecting the antibody; d) scoring the detection pathologically, wherein a pathology score of 0 indicates no staining in the sample, a score of 1+ indicates weak staining in the sample; a score of 2+ indicates intermediate staining in the sample; and a score of 3+ indicates strong staining in the sample; and e) providing a prognosis to the patient, wherein the pathology score inversely correlates with the severity of disease.