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Åke Lernmark

Principal investigator

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Comparison of three assays for the detection of GAD65Ab-specific anti-idiotypic antibodies

Author

  • Shilpa Oak
  • Jared Radtke
  • Mona Landin-Olsson
  • Carina Törn
  • Åke Lernmark
  • Christiane S. Hampe

Summary, in English

Anti-idiotypic antibodies (anti-Id) to autoantibodies are present in several autoimmune diseases and are hypothesized to have regulatory function. Recently we reported the presence of anti-Id to a major type 1 diabetes-associated autoantibody (CAD65Ab) in sera of healthy individuals. Our current assay for the detection of GAD65Ab-specific anti-Id requires the initial removal of anti-Id from the sera using immobilized monoclonal GAD65Ab, followed by detection of the now exposed GAD65Ab. However, anti-Id in samples that are GAD65Ab-negative cannot be detected in this assay. Furthermore, we cannot distinguish between serum GAD65Ab and the monoclonal GAD65Ab used in the absorption of anti-Id. In this study we evaluated two novel detection assays for GAD65Ab-specific anti-Id. The biotin/streptavidin based absorption assay utilizes the strong interaction of biotin and streptavidin to prevent possible leakage of the immobilized antibody. Moreover, this assay format allows to identify the origin of the detected GAD65Ab. The ECL-based assay allows the direct detection of anti-Id independent of the presence of GAD65Ab. We analyzed new-onset type 1 diabetes patients (n = 133) and matched healthy controls (n = 178) for the presence of GAD65Ab-specific anti-Id using both new detection assays and the original absorption assay. We found that all three assays can distinguish between the type 1 diabetes cohort and the healthy control samples. The biotin/streptavidin assay allowed us to positively exclude the monoclonal GAD65Ab as the source of the detected GAD65Ab. While the original absorption assay showed the highest sensitivity and specificity, the ECL format showed the highest peak signal-to-noise ratio and excellent linear correlation, making this assay our first choice for quantification of anti-Id. (C) 2009 Elsevier B.V. All rights reserved.

Department/s

  • Medicine, Lund
  • Diabetes and Celiac Unit

Publishing year

2009

Language

English

Pages

55-61

Publication/Series

Journal of Immunological Methods

Volume

351

Issue

1-2

Document type

Journal article

Publisher

Elsevier

Topic

  • Immunology in the medical area

Keywords

  • Autoantibodies
  • decarboxylase
  • Glutamate
  • Anti-idiotypic antibodies
  • Radioligand binding assay
  • Type 1 diabetes
  • ECL

Status

Published

Research group

  • Diabetes and Celiac Unit

ISBN/ISSN/Other

  • ISSN: 1872-7905