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Front Neurol. 2017 Jul 6;8:305. doi: 10.3389/fneur.2017.00305. eCollection 2017.

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

Author information

1
Medical Faculty, Department of Neurology, Heinrich-Heine-University, Duesseldorf, Germany.
2
Sanofi-Aventis, Deutschland GmbH, Frankfurt am Main, Germany.
3
Neuroimmunology Laboratory, DMSC, Department of Neurology, Rigshospitalet, Copenhagen, Denmark.
4
Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden.
5
Department of Neurology, Innsbruck Medical University, Innsbruck, Austria.
6
INSERM 996, University Paris-Sud, Paris, France.
7
Merck NBE Bioanalytics, Torino, Italy.
8
Centre for Neuroscience and Trauma, Blizard Institute, Queen Mary, University of London, London, United Kingdom.
9
Department of Neurology, University Hospital of Cologne, Cologne, Germany.

Abstract

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.

KEYWORDS:

anti-drug antibodies; biotherapy; enzyme-linked immunosorbent assay; interferon beta; multiple sclerosis

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