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Carina Törn

Coordinator

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Determination of glutamic acid decarboxylase antibodies (GADA) IgG subclasses - comparison of three immunoprecipitation assays (IPAs)

Author

  • Magnus Hillman
  • Carina Törn
  • Mona Landin-Olsson

Summary, in English

IgG subclasses of glutamic acid decarboxylase (GAD(65)) antibodies (GADA) may reflect the immunological state in the pancreas of GADA-positive patients with autoimmune diabetes. The use of biotin-conjugated antibodies and streptavidin Sepharose are used commonly in immunoprecipitation assays (IPA) based on (125)I- or (35)S-labelled antigens to capture IgG subclasses directed against IA-2 or GAD(65). We have compared three different immunoprecipitation assays for the determination of GADA IgG subclasses. Two of the assays were based on the biotin and streptavidin systems provided in a solid (immobilized) or liquid (mobilized) phase binding environment. The third assay was based on N-hydroxysuccinimide (immobilized) interaction with primary amines (i.e. lysine residues) on the antibody. We found the liquid phase binding assay (LPBA) to be the most stable assay, with a comparatively low coefficient of variation and background.

Department/s

  • Medicine, Lund
  • Diabetes and Celiac Unit

Publishing year

2007

Language

English

Pages

68-74

Publication/Series

Clinical and Experimental Immunology

Volume

150

Issue

1

Document type

Journal article

Publisher

British Society for Immunology

Topic

  • Immunology in the medical area

Status

Published

Research group

  • Diabetes and Celiac Unit

ISBN/ISSN/Other

  • ISSN: 0009-9104