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

Clinical Department of Neurology, Innsbruck Medical University, Anichstrasse, Innsbruck, Austria

M Brugger

Clinical Department of Neurology, Innsbruck Medical University, Anichstrasse, Innsbruck, Austria

A Millonig

Clinical Department of Neurology, Innsbruck Medical University, Anichstrasse, Innsbruck, Austria

A Fogdell-Hahn

Division of Neurology, Karolinska Institutet, Clinical Neuroscience, Karolinska University Hospital, Huddinge, Stockholm, Sweden

D Rudzki

Clinical Department of Neurology, Innsbruck Medical University, Anichstrasse, Innsbruck, Austria

J Hillert

Division of Neurology, Karolinska Institutet, Clinical Neuroscience, Karolinska University Hospital, Huddinge, Stockholm, Sweden

T Berger

Clinical Department of Neurology, Innsbruck Medical University, Anichstrasse, Innsbruck, Austria

M Reindl

Clinical Department of Neurology, Innsbruck Medical University, Anichstrasse, Innsbruck, Austria

F Deisenhammer

Clinical Department of Neurology, Innsbruck Medical University, Anichstrasse, Innsbruck, Austria, florian.deisenhammer{at}i-med.ac.at

Background

Binding antibodies (BAB) against interferon-β (IFNβ) 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.

Key Words: antibodies • binding • enzyme-linked immunosorbent assay • interferon-β • multiple sclerosis • neutralizing • western blot

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