Title |
Development and Validation of an Enzyme-Linked Immunosorbent Assay for the Detection of Binding Anti-Drug Antibodies against Interferon Beta
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Published in |
Frontiers in Neurology, July 2017
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DOI | 10.3389/fneur.2017.00305 |
Pubmed ID | |
Authors |
Kathleen Ingenhoven, Daniel Kramer, Poul Erik 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 |
Abstract |
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. 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. 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. 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. |
X Demographics
Geographical breakdown
Country | Count | As % |
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Switzerland | 1 | 50% |
Unknown | 1 | 50% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 2 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 21 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 4 | 19% |
Student > Bachelor | 2 | 10% |
Other | 2 | 10% |
Student > Postgraduate | 2 | 10% |
Student > Master | 2 | 10% |
Other | 4 | 19% |
Unknown | 5 | 24% |
Readers by discipline | Count | As % |
---|---|---|
Immunology and Microbiology | 6 | 29% |
Biochemistry, Genetics and Molecular Biology | 4 | 19% |
Agricultural and Biological Sciences | 2 | 10% |
Medicine and Dentistry | 2 | 10% |
Chemistry | 1 | 5% |
Other | 1 | 5% |
Unknown | 5 | 24% |