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Method for detecting autoimmune antibodies against TSH receptors and novel TSH-receptor chimaeras

An autoimmune antibody, autoantibody technology, applied in receptors/cell surface antigens/cell surface determinants, hormone receptors, biological tests, etc., can solve problems such as inability to distinguish irritants and inability to identify subtypes

Active Publication Date: 2008-12-10
乌尔里希洛斯
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disadvantage is the inability to distinguish stimulatory, blocking and neutral autoimmune antibodies
Furthermore, not all isoforms could be identified as this substituted TSH only binds 30-40% to epitopes for stimulatory and blocking autoantibodies

Method used

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  • Method for detecting autoimmune antibodies against TSH receptors and novel TSH-receptor chimaeras
  • Method for detecting autoimmune antibodies against TSH receptors and novel TSH-receptor chimaeras
  • Method for detecting autoimmune antibodies against TSH receptors and novel TSH-receptor chimaeras

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Experimental program
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Embodiment

[0098] material

[0099] The plasmid pcDNA3-rLHR (B9) was provided by Dr. D.L. Segaloff (The University of Iowa, USA). The plasmid pSP-luc-NF was obtained from Promega GmbH (Heidelberg, Deutschland). The kit of Amersham GmbH (Braunschweig, Deutschland) was used as the ECL Western blotting kit and the cAMP RIA kit.

[0100] Used CLONTECH Laboratories, Inc (Palo Alto, CA, USA) pIRESneo (expression vector), pSEAP2-Basic, PJKGmbH (Kleinbittersdorf, Deutschland) Gaussia luciferase, Laboratory voor Monoklonale Antistoffen (LMA) (Wageningen, Niederlande) The alkaline phosphatase Escherichia coli, SYNTHETIC GENE, horseradish peroxidase of horseradish (Armoracia rustician) of British Bio-Technology Ltd (UK), human thyroid stimulating hormone (Nakabayashi et al., "Thyrostimulin, a heterodimer of two newhuman glycoprotein hormone subunits, activates the thyroid-stimulating hormone receptor", J. Clin. 109(11), 1445-1452).

[0101] DNA primer

[0102] P1(5’-GTCATGCATCAGCTGCTGGTGCTGGCAGTG-3’)...

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Abstract

The invention relates to a method for identifying autoimmune antibodies against TSH receptors with the aid of TSH receptor chimaeras, which, preferably, contain chimaeras which are embodied only in the form of the modified extracellular fraction of a TSH savage-type receptor and are modified by highly immunogenic peptide groups or by detection of suitable enzymes, wherein the inventive determining method makes it possible to easily detecting stimulating, blocking and neutral autoimmune antibodies.

Description

Invention field [0001] The present invention relates to a method for highly specific determination of different types of autoantibodies against thyroid stimulating hormone receptor (TSH receptor), and a novel TSH receptor chimera that can be used as a binding agent in the method. Background of the invention [0002] The TSH receptor (TSH-R) plays a key role in the function and growth of thyroid cells. This receptor is a member of the G protein-coupled glycoprotein receptor subfamily, which also specifically includes the luteinizing hormone / chorionic gonadotropin receptor (LH / CGR) and the follicle-stimulating hormone receptor ( FSHR). The receptors of this subfamily have a large N-terminal extracellular domain, which is very important for ligand binding, and therefore appears to be involved in signal transmission for them. The transmission of TSHR signal is mainly mediated by activation of adenylate cyclase, and activation of adenylate cyclase leads to an increase in intracellular...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/76G01N33/564C07K14/72
CPCC07K14/723G01N33/564G01N33/76G01N33/53
Inventor 乌尔里希·洛斯
Owner 乌尔里希洛斯
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