Detection of antibodies directed to the N-terminal region of GAD is dependent on assay format and contributes to differences in the specificity of GAD autoantibody assays for type 1 diabetes

Alistair J K Williams, Vito Lampasona, Michael Schlosser, Patricia W. Mueller, David L. Pittman, William E. Winter, Beena Akolkar, Rebecca Wyatt, Cristina Brigatti, Stephanie Krause, Peter Achenbach

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

GAD autoantibodies (GADAs) are sensitive markers of islet autoimmunity and type 1 diabetes. They form the basis of robust prediction models and are widely used for the recruitment of subjects at high risk of type 1 diabetes to prevention trials. However, GADAs are also found in many individuals at low risk of diabetes progression. To identify the sources of diabetes-irrelevant GADA reactivity, we analyzed data from the 2009 and 2010 Diabetes Autoantibody Standardization Program GADA workshop and found that binding of healthy control sera varied according to assay type. The characterization of control sera found positive by radiobinding assay (RBA), but negative by ELISA, showed that many of these sera reacted to epitopes in the N-terminal region of the molecule. This finding prompted development of an N-terminally truncated GAD65 radiolabel, 35S-GAD65(96-585), which improved the performance of most GADA RBAs participating in an Islet Autoantibody Standardization Program GADA substudy. These detailed workshop comparisons have identified a source of disease-irrelevant signals in GADA RBAs and suggest that N-terminally truncated GAD labels will enable more specific measurement of GADAs in type 1 diabetes.

Original languageEnglish
Pages (from-to)3239-3246
Number of pages8
JournalDiabetes
Volume64
Issue number9
DOIs
Publication statusPublished - Sep 1 2015

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Type 1 Diabetes Mellitus
Autoantibodies
Antibodies
Serum
Education
Autoimmunity
Epitopes
Enzyme-Linked Immunosorbent Assay

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism

Cite this

Detection of antibodies directed to the N-terminal region of GAD is dependent on assay format and contributes to differences in the specificity of GAD autoantibody assays for type 1 diabetes. / Williams, Alistair J K; Lampasona, Vito; Schlosser, Michael; Mueller, Patricia W.; Pittman, David L.; Winter, William E.; Akolkar, Beena; Wyatt, Rebecca; Brigatti, Cristina; Krause, Stephanie; Achenbach, Peter.

In: Diabetes, Vol. 64, No. 9, 01.09.2015, p. 3239-3246.

Research output: Contribution to journalArticle

Williams, AJK, Lampasona, V, Schlosser, M, Mueller, PW, Pittman, DL, Winter, WE, Akolkar, B, Wyatt, R, Brigatti, C, Krause, S & Achenbach, P 2015, 'Detection of antibodies directed to the N-terminal region of GAD is dependent on assay format and contributes to differences in the specificity of GAD autoantibody assays for type 1 diabetes', Diabetes, vol. 64, no. 9, pp. 3239-3246. https://doi.org/10.2337/db14-1693
Williams, Alistair J K ; Lampasona, Vito ; Schlosser, Michael ; Mueller, Patricia W. ; Pittman, David L. ; Winter, William E. ; Akolkar, Beena ; Wyatt, Rebecca ; Brigatti, Cristina ; Krause, Stephanie ; Achenbach, Peter. / Detection of antibodies directed to the N-terminal region of GAD is dependent on assay format and contributes to differences in the specificity of GAD autoantibody assays for type 1 diabetes. In: Diabetes. 2015 ; Vol. 64, No. 9. pp. 3239-3246.
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