A New Luminescence Assay for Autoantibodies to Mammalian Cell-Prepared IA-2
- Peter D. Burbelo, Ph.D.1,
- Hiroki Hirai, M.D., Ph.D.2,
- Hannah Leahy1,
- Ake Lernmark, M.D.3,
- Sten-A. Ivarsson, M.D.4,
- Michael J. Iadarola, Ph.D. (miadarola{at}mail.nih.gov)1 and
- Abner L. Notkins, M.D. (anotkins{at}mail.nih.gov)2
- 1Sensory Biology Branch, and
- 2Oral Infection and Immunity Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD, 20892, USA
- 3Department of Medicine, R. H. Williams Laboratory, University of Washington, Seattle, WA, USA
- 4Department of Clinical Sciences, University Hospital MAS, Lund University, Malmö, Sweden
Abstract
Objective: IA-2 is a major autoantigen in type 1 diabetes and autoantibodies to it are routinely detected by a liquid phase radioimmunoprecipitation assay. The present experiments were initiated to develop a new assay that does not require the use of radioisotopes or autoantigens prepared in bacteria or by in vitro transcription/translation.
Research Design and Methods: IA-2 luciferase fusion protein was expressed in mammalian cells and assayed for autoantibodies by liquid phase luciferase immunoprecipitation.
Results: Our study showed that there was no significant difference between the luciferase immunoprecipitation and the radioimmunoprecipitation assays in sensitivity and specificity, and comparison of the two assays revealed a high correlation coefficient (R2 = 0.805).
Conclusion: The luciferase system offers a robust, inexpensive, non-radioactive method for the detection of autoantibodies to mammalian cell-prepared IA-2 and should be of practical value at the clinical level.
Footnotes
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- Received February 19, 2008.
- Accepted May 23, 2008.
- Copyright © American Diabetes Association
