Comparative study of four different assays for the detection of binding antibodies against interferon-beta

Mult Scler. 2008 Jul;14(6):830-6. doi: 10.1177/1352458508089228. Epub 2008 Jun 5.

Abstract

Background: Binding antibodies (BAB) against interferon-beta (IFNbeta) are often determined as screening assays before performing an expensive and elaborate neutralizing antibody (NAB) test.

Methods: In this study, we compared four BAB tests, a western blot (WB), a direct binding enzyme-linked immunosorbent assay (ELISA) (dELISA), a capture ELISA (cELISA), and a commercial enzyme immuno-assay (EIA) in 325 multiple sclerosis patients with and without neutralizing antibodies to evaluate the sensitivity and specificity to detect NAB by receiver operating characteristics analysis.

Results: The area under the curve (AUC) values were 0.907 for the dELISA, 0.925 for the cELISA, and 0.776 for the EIA (P < 0.0001 for all). At a sensitivity of 95%, the specificity was approximately 30% in the dELISA, 55% in the cELISA, and 13% in the EIA. The WB as a qualitative BAB detection method had a given sensitivity of 97% and a specificity of 55%. There was a strong and significant correlation between high NAB titers (>500 neutralizing units [NU]) and titers obtained by all quantitative BAB assays. However, low to medium NAB titers (20-500 NU) did not significantly correlate with BAB titers.

Conclusion: We conclude that the cELISA seems to be most suitable for NAB screening, but BAB titers cannot reliably predict NAB titers.

Publication types

  • Comparative Study
  • Validation Study

MeSH terms

  • Antibodies / isolation & purification*
  • Blotting, Western / standards
  • Cell Line, Tumor
  • Enzyme-Linked Immunosorbent Assay / methods
  • Enzyme-Linked Immunosorbent Assay / standards*
  • Humans
  • Immunoassay / standards
  • In Vitro Techniques
  • Interferon-beta / immunology*
  • Lung Neoplasms
  • Mass Screening
  • Multiple Sclerosis, Relapsing-Remitting / drug therapy*
  • Multiple Sclerosis, Relapsing-Remitting / immunology*
  • Neutralization Tests
  • ROC Curve
  • Reagent Kits, Diagnostic / standards
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • Antibodies
  • Reagent Kits, Diagnostic
  • Interferon-beta