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Development and Validation of an Enzyme-Linked Immunosorbent Assay for the Detection of Binding Anti-Drug Antibodies against Interferon Beta

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  • Kathleen Ingenhoven
  • Daniel Kramer
  • Poul Erik Hyldgaard Jensen
  • Christina Hermanrud
  • Malin Ryner
  • Florian Deisenhammer
  • Marc Pallardy
  • Til Menge
  • Hans-Peter Hartung
  • Bernd C Kieseier
  • Elisa Bertotti
  • Paul Creeke
  • Anna Fogdell-Hahn
  • Clemens Warnke
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OBJECTIVE: To develop and validate a method for the detection of binding anti-drug antibodies (ADAs) against interferon beta (IFN-β) in human serum as part of a European initiative (ABIRISK) aimed at the prediction and analysis of clinical relevance of anti-biopharmaceutical immunization to minimize the risk.

METHOD: A two-tiered bridging enzyme-linked immunosorbent assay (ELISA) format was selected and validated according to current recommendations. Screening assay: ADA in serum samples form complexes with immobilized IFN-β and biotinylated IFN-β, which are then detected using HRP labeled Streptavidin and TMB substrate. Confirmation assay: Screen "putative positive" samples are tested in the presence of excess drug (preincubation of sera with 0.3 µg/mL of soluble IFN-β) and percentage of inhibition is calculated.

RESULTS: The assay is precise, and the sensitivity of the assay was confirmed to be 26 ng/mL using commercially available polyclonal rabbit antihuman IFN-β in human sera as the positive control.

CONCLUSION: An ultrasensitive ELISA for IFN-β-binding ADA testing has been validated. This will form the basis to assess anti-biopharmaceutical immunization toward IFN-β with regards to its clinical relevance and may allow for the development of predictive tools, key aims within the ABIRISK consortium.

Original languageEnglish
JournalFrontiers in Neurology
Volume8
Pages (from-to)305
Number of pages10
ISSN1664-2295
DOIs
Publication statusPublished - 2017

    Research areas

  • Journal Article

ID: 52217161