A simple and fast non-radioactive bridging immunoassay for insulin autoantibodies

PLoS One. 2013 Jul 29;8(7):e69021. doi: 10.1371/journal.pone.0069021. Print 2013.

Abstract

Type 1 diabetes (T1D) is an autoimmune disease which results from the destruction of pancreatic beta cells. Autoantibodies directed against islet antigens are valuable diagnostic tools. Insulin autoantibodies (IAAs) are usually the first to appear and also the most difficult to detect amongst the four major islet autoantibodies. A non-radioactive IAA bridging ELISA was developed to this end. In this assay, one site of the IAAs from serum samples is bound to a hapten-labeled insulin (GC300-insulin), which is subsequently captured on anti-GC300 antibody-coated 96-well plates. The other site of the IAAs is bound to biotinylated insulin, allowing the complex to be detected by an enzyme-streptavidin conjugate. In the present study, 50 serum samples from patients with newly diagnosed T1D and 100 control sera from non-diabetic individuals were analyzed with our new assay and the results were correlated with an IAA radioimmunoassay (RIA). Using IAA bridging ELISA, IAAs were detected in 32 out of 50 T1D children, whereas with IAA RIA, 41 out of 50 children with newly diagnosed T1D were scored as positive. In conclusion, the IAA bridging ELISA could serve as an attractive approach for rapid and automated detection of IAAs in T1D patients for diagnostic purposes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Antibodies, Monoclonal / blood
  • Autoantibodies / blood*
  • Child
  • Child, Preschool
  • Electrochemical Techniques
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Female
  • Humans
  • Infant
  • Infant, Newborn
  • Insulin Antibodies / blood*
  • Luminescent Measurements
  • Male
  • ROC Curve
  • Radioimmunoassay / methods*
  • Sensitivity and Specificity

Substances

  • Antibodies, Monoclonal
  • Autoantibodies
  • Insulin Antibodies

Grants and funding

Ingrid Kikkas obtained a grant from the PhD programme of the Commisariat à l'Energie Atomique. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.