# US 9,518,993 B2 · Reagents and methods for detecting a polymorphic protein > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,518,993 B2 - Document ID: US9518993B2 - Status: Granted - Kind: B2 - Application number: 14/530,836 - Filed: 2014-11-03 - Published: 2015-02-26 - Granted: 2016-12-13 - Earliest claimed: 2008-06-13 - Assignees: Rappaport Family Institute for Research in the Medical Sciences - Applicants: Rappaport Family Institute for Research In the Medical Sciences - Inventors: Jacob Victor; Noah Berkowitz; Andrew Levy - Web page: https://patentgrep.com/patent/US9518993B2 ## CPC - G01N 33/6893: {related to diseases not provided for elsewhere} - C07K 16/18: against material from animals or humans - C07K 2317/56: variable (Fv) region, i.e. VH and/or VL - C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value - G01N 2800/042: Disorders of carbohydrate metabolism, e.g. diabetes, glucose metabolism - 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) - 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 antibodies that differentially react with allelic variants of a polymorphic protein, methods of identifying same, an antigen binding fragment comprised therein, proteins, cells, viral particles, compositions, and kits comprising same. The invention also provides methods for determining a haptoglobin type of a subject and methods for testing a subject for susceptibility to diabetic complications. ## Claims ### Claim 1 A method of determining a haptoglobin (Hp) phenotype of a subject, comprising: contacting a biological sample of said subject with an isolated anti-haptoglobin monoclonal antibody, wherein the isolated anti-haptoglobin monoclonal antibody, comprises at least one light chain variable region and at least one heavy chain variable region, said light chain variable region comprising: the complementarity determining region light chain (CDRL) 1 consisting of SEQ ID NO:3; a CDRL2 consisting of SEQ ID NO:4; and a CDRL3 consisting of SEQ ID NO:5, and said heavy chain variable region comprising: a complementarity determining region heavy chain (CDRH) 1 consisting of SEQ ID NO:8; a CDRH2 consisting of SEQ ID NO:9; and a CDRH3 consisting of SEQ ID NO:10; forming a bound complex between the isolated anti-haptoglobin monoclonal antibody and haptoglobin in the biological sample; quantitatively determining a binding affinity between said haptoglobin and said isolated anti-haptoglobin monoclonal antibody; and comparing said quantitatively determined binding affinity with a value obtained from quantitatively determining the binding affinity of said isolated anti-haptoglobin monoclonal antibody to an isolated Hp 1-1, Hp 2-1, or Hp 2-2 isoform, wherein the binding affinity determined is indicative of the Hp allele type, thereby determining the phenotype of Hp in said subject. ### Claim 2 (depends on 1) The method of claim 1, wherein the biological sample is serum or plasma. ### Claim 3 (depends on 2) The method of claim 2, wherein the plasma has been anticoagulated with citrate, heparin or EDTA. ### Claim 4 (depends on 1) The method of claim 1, further comprising immobilizing the bound anti-haptoglobin antibody-haptoglobin complex on a substrate resulting in an immobilized complex. ### Claim 5 (depends on 4) The method of claim 4, wherein the immobilizing comprises binding the anti-haptoglobin monoclonal antibody to the substrate. ### Claim 6 (depends on 4) The method of claim 4, further comprising: contacting said immobilized complex with an additional quantity of said monoclonal anti-haptoglobin antibody; and determining a binding affinity between said haptoglobin and said additional quantity of said monoclonal anti-haptoglobin antibody. ### Claim 7 (depends on 1) The method of claim 1, wherein the monoclonal anti-haptoglobin antibody is a murine hybridoma deposited with the American Type Culture Collection (ATCC) having a patent deposit designation PTA-9815. ### Claim 8 (depends on 1) The method of claim 1, wherein the isolated anti-haptoglobin monoclonal antibody comprises a light chain variable region consisting of SEQ ID NO:2. ### Claim 9 (depends on 1) The method of claim 1, wherein the isolated anti-haptoglobin monoclonal antibody comprises a heavy chain variable region consisting of SEQ ID NO:7. ### Claim 10 (depends on 1) The method of claim 1, wherein the isolated anti-haptoglobin monoclonal antibody comprises a light chain variable region consisting of SEQ ID NO:2 and a heavy chain variable region consisting of SEQ ID NO:7. ### Claim 11 (depends on 1) The method of claim 1, wherein said antibody light or heavy chain variable region of the isolated anti-haptoglobin monoclonal antibody is at least one of chimerzed, humanized, or CDR-grafted. ### Claim 12 (depends on 11) The method of claim 11, wherein the isolated anti-haptoglobin monoclonal antibody further comprises at least one compound or polypeptide selected from a detectable label or reporter. ### Claim 13 (depends on 12) The method of claim 12, wherein the detectable label is an enzyme. ### Claim 14 (depends on 13) The method of claim 13, wherein the enzyme is horseradish peroxidase or alkaline phosphatase. ### Claim 15 (depends on 1) The method of claim 1, wherein the isolated anti-haptoglobin monoclonal antibody has a binding affinity for Hp2-2, a binding affinity for Hp2-1, and a binding affinity for Hp1-1, wherein the binding affinity of the isolated anti-haptoglobin monoclonal antibody for Hp2-2 is greater than the binding affinity of the isolated anti-haptoglobin monoclonal antibody for Hp2-1 and Hp-1-1, and the binding affinity of the isolated anti-haptoglobin monoclonal antibody for Hp2-1 is greater than the binding affinity of the isolated anti-haptoglobin monoclonal antibody for Hp-1-1.