# US 8,999,727 B2 · Innovative TSH-R-Ab-kit > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,999,727 B2 - Document ID: US8999727B2 - Status: Granted - Kind: B2 - Application number: 11/992,563 - Filed: 2006-09-25 - Published: 2009-12-31 - Granted: 2015-04-07 - Earliest claimed: 2005-09-26 - Assignees: Ulrich Loos - Applicants: Ulrich Loos - Inventors: Ulrich Loos - Web page: https://patentgrep.com/patent/US8999727B2 ## CPC - G01N 33/76: Human chorionic gonadotropin {including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors} - C07K 14/723: {G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor} - G01N 33/53: Immunoassay; Biospecific binding assay; Materials therefor - G01N 33/564: for pre-existing immune complex or autoimmune disease {, i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9} - 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 Methods are described for detection of autoimmune antibodies against the TSH receptor using TSH receptor chimeras, which preferably only contain the extracellular portion of the TSH wild type receptor modified as chimera, and are modified by highly immunogenic peptide residues or by enzymes suitable for detection, wherein the determination methods described allow simple detection of stimulating, blocking and neutral autoimmune antibodies. ## Claims ### Claim 1 A method for determination of stimulating autoimmune antibodies directed against the thyroid-stimulating hormone (TSH) receptor in a patient sample, using TSH receptor chimera B, wherein the amino acids from 261 to 370 of the TSH receptor are replaced by the comparable amino acids from 261 to 329 of a rat LH-CGR said method comprising: (a) contacting a patient sample that is to be tested for a stimulating autoimmune antibody with a first receptor chimera wherein said first receptor chimera B is a holoreceptor chimera is bound to a solid phase, and wherein said stimulating autoimmune antibody comprises a first antigen-binding site and a second antigen-binding site; (b) allowing binding of said stimulating autoimmune antibody first antigen-binding site with said first receptor chimera B; (c) adding a second receptor chimera B, wherein said second receptor chimera B is a truncated chimera B, wherein said truncated chimera B comprises only the extracellular domain of said first receptor chimera B, and wherein said second receptor chimera B is modified at its C-terminal end or N-terminal end to allow for the detection of said second receptor chimera B; (d) allowing binding of said stimulating autoimmune antibody second antigen-binding site to said second receptor chimera B; and (e) determining if a stimulating autoimmune antibody is present in said patient sample. ### Claim 2 (depends on 1) The method according to claim 1, wherein said first receptor chimera B is bound to said solid phase by an immobilized antibody, and said second receptor chimera B does not bind to said immobilized antibody. ### Claim 3 (depends on 2) The method according to claim 2, wherein said second receptor chimera B is modified at its C-terminal end by fusion to a peptide that is recognized by a secondary antibody suitable for detection. ### Claim 4 (depends on 3) The method according to claim 3, wherein said peptide is a subsequence from thyrostimulin. ### Claim 5 (depends on 2) The method according to claim 2, wherein said second receptor chimera B is modified at its C-terminal end with a label suitable for detection. ### Claim 6 (depends on 2) The method according to claim 2, wherein said first receptor chimera B is modified at its C-terminal end by fusion to a peptide that is recognized by said immobilized antibody. ### Claim 7 (depends on 6) The method according to claim 6, wherein the peptide is a subsequence from thyrostimulin. ### Claim 8 (depends on 5) The method according to claim 5, wherein said label is an acridine compound, a ruthenium compound, biotin, streptavidin or fluorescein isothiocyanate. ### Claim 9 (depends on 5) The method according to claim 5, wherein said label is a secretory alkaline phosphatase, a glowworm luciferase, a Gaussia luciferase, or a peroxidase. ### Claim 10 (depends on 1) The method according to claim 1, wherein the second receptor chimera B is modified at its C-terminal end. ### Claim 11 (depends on 1) The method according to claim 1, wherein the second receptor B chimera is modified at its N-terminal end. ### Claim 12 (depends on 2) The method according to claim 2, wherein the second receptor chimera B is modified at its N-terminal end by fusion to a peptide that is recognized by a secondary antibody suitable for detection. ### Claim 13 (depends on 12) The method according to claim 12, wherein said peptide is a subsequence from thyrostimulin. ### Claim 14 (depends on 2) The method according to claim 2, wherein said second receptor chimera B is modified at its C-terminal end by fusion to an enzyme. ### Claim 15 (depends on 14) The method of claim 14, wherein said enzyme is alkaline phosphatase, secretory alkaline phosphatase, a glowworm luciferase, a Gaussia luciferase, or a peroxidase. ### Claim 16 (depends on 2) The method according to claim 2, wherein said second receptor chimera B is modified at its N-terminal end by fusion to an enzyme. ### Claim 17 (depends on 16) The method of claim 16, wherein said enzyme is alkaline phosphatase, secretory alkaline phosphatase, a glowworm luciferase, a Gaussia luciferase, or a peroxidase.