A New Luminescence Assay for Autoantibodies to Mammalian Cell–Prepared Insulinoma-Associated Protein 2

  1. Peter D. Burbelo, PHD1,
  2. Hiroki Hirai, MD, PHD2,
  3. Hannah Leahy, BA1,
  4. Ake Lernmark, MD3,
  5. Sten A. Ivarsson, MD4,
  6. Michael J. Iadarola, PHD1 and
  7. Abner Louis Notkins, MD2
  1. 1Sensory Biology Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland
  2. 2Oral Infection and Immunity Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland
  3. 3Department of Medicine, R.H. Williams Laboratory, University of Washington, Seattle, Washington
  4. 4Department of Clinical Sciences, University Hospital MAS, Lund University, Malmö, Sweden
  1. Corresponding authors: Abner Louis Notkins, anotkins{at}mail.nih.gov and Michael J. Iadarola, miadarol{at}mail.nih.gov

Abstract

OBJECTIVE—Insulinoma-associated protein 2 (IA-2) is a major autoantigen in type 1 diabetes, and IA-2 autoantibodies 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).

CONCLUSIONS—The luciferase system offers a robust, inexpensive, nonradioactive method for the detection of autoantibodies to mammalian cell–prepared IA-2 and could be of practical value at the clinical level.

Footnotes

  • Published ahead of print at http://care.diabetesjournals.org on 5 June 2008.

    P.D.B. and H.H. contributed equally to this work.

    P.D.B. has filed a patent application based on LIPS technology.

    Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for details.

    The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C Section 1734 solely to indicate this fact.

    • Accepted May 23, 2008.
    • Received February 19, 2008.
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  1. Published online before print June 5, 2008, doi: 10.2337/dc08-0286 Diabetes Care September 2008 vol. 31 no. 9 1824-1826
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