IngentaConnect Determination of glutamic acid decarboxylase antibodies (GADA) Ig...

Authors: Hillman, M.; Törn, C.¹; Landin-Olsson, M.²

Source: Clinical & Experimental Immunology, Volume 150, Number 1, October 2007 , pp. 68-74(7)

Abstract:

Summary

IgG subclasses of glutamic acid decarboxylase (GAD₆₅) 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 ¹²⁵I- or ³⁵S-labelled antigens to capture IgG subclasses directed against IA-2 or GAD₆₅. 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.

Keywords: anti-glutamate decarboxylase antibodies; autoantibodies; autoimmunity; diabetes; isotype switching; isotypes

Document Type: Research article

DOI: 10.1111/j.1365-2249.2007.03473.x

Affiliations: 1: Unit for Diabetes and Coeliac Disease, CRC, Malmö University Hospital, Malmö, Sweden 2: Diabetes Research Laboratory, Lund University Hospital, Lund, Sweden, and

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