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

  1. M. Hillman1,2,*,
  2. C. Törn3,
  3. M. Landin-Olsson2

Article first published online: 30 JUL 2007

DOI: 10.1111/j.1365-2249.2007.03473.x

Issue

Clinical & Experimental Immunology

Clinical & Experimental Immunology

Volume 150, Issue 1, pages 68–74, October 2007

Additional Information

How to Cite

Hillman, M., Törn, C. and Landin-Olsson, M. (2007), Determination of glutamic acid decarboxylase antibodies (GADA) IgG subclasses − comparison of three immunoprecipitation assays (IPAs). Clinical & Experimental Immunology, 150: 68–74. doi: 10.1111/j.1365-2249.2007.03473.x

Author Information

  1. 1

    Department of Clinical Sciences, Lund University,

  2. 2

    Diabetes Research Laboratory, Lund University Hospital, Lund, Sweden, and

  3. 3

    Unit for Diabetes and Coeliac Disease, CRC, Malmö University Hospital, Malmö, Sweden

*Magnus Hillman, Diabetes Research Laboratory, B11, BMC, 221 84 Lund, Sweden. E-mail: Magnus.Hillman@med.lu.se

Publication History

  1. Issue published online: 30 JUL 2007
  2. Article first published online: 30 JUL 2007
  3. Accepted for publication 21 June 2007

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Keywords:

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

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.

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