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

Abstract

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.

Keywords

antibodies binding enzyme-linked immunosorbent assay interferon-β multiple sclerosis neutralizing western blot

Get full access to this article

View all access options for this article.

References

1 The IFNb multiple sclerosis study group, The University of British Columbia MS/MRI analysis group. Interferon beta-1b in the treatment of multiple sclerosis: final outcome of the randomized controlled trial. Neurology 1995; 45: 1277 – 1285 .

2 Jacobs LD Cookfair DL Rudick RA . A phase III trial of intramuscular recombinant interferon beta as treatment for exacerbating-remitting multiple sclerosis: design and conduct of study and baseline characteristics of patients. Multiple Sclerosis Collaborative Research Group (MSCRG). Mult Scler 1995; 1: 118 – 135 .

3 PRISMS . Randomised double-blind placebo-controlled study of interferon β-1a in relapsing/remitting multiple sclerosis. Lancet 1998; 352: 1498 – 1504 .

4 Goodin DS Frohman EM Hurwitz B . Neutralizing antibodies to interferon beta: assessment of their clinical and radiographic impact: an evidence report: report of the Therapeutics and Technology Assessment Subcommittee of the American Academy of Neurology. Neurology 2007; 68: 977 – 984 .

5 Sorensen PS Deisenhammer F Duda P . Guidelines on use of anti-IFNb antibody measurements in multiple sclerosis: report of an EFNS Task Force on IFNb antibodies in multiple sclerosis. Eur J Neurol 2005; 12: 817 – 827 .

6 Deisenhammer F Schellekens H Bertolotto A . Measurement of neutralizing antibodies to interferon beta in patients with multiple sclerosis. J Neurol 2004; 251(Suppl. 2): II31 – II39 .

7 Namaka M Pollitt-Smith M Gupta A . The clinical importance of neutralizing antibodies in relapsing–remitting multiple sclerosis. Curr Med Res Opin 2006; 22: 223 – 239 .

8 Sorensen PS Ross C Clemmesen KM . Clinical importance of neutralising antibodies against interferon beta in patients with relapsing–remitting multiple sclerosis. Lancet 2003; 362: 1184 – 1191 .

9 Sorensen PS Tscherning T Mathiesen HK . Neutralizing antibodies hamper IFNbeta bioactivity and treatment effect on MRI in patients with MS. Neurology 2006; 67: 1681 – 1683 .

10.

10 Francis GS Rice GP Alsop JC . Interferon beta-1a in MS: results following development of neutralizing antibodies in PRISMS. Neurology 2005; 65: 48 – 55 .

11.

11 Deisenhammer F Reindl M Harvey J Gasse T Dilitz E Berger T . Bioavailability of interferon beta 1b in MS patients with and without neutralizing antibodies. Neurology 1999; 52: 1239 – 1243 .

12.

12 Bertolotto A Gilli F Sala A . Persistent neutralizing antibodies abolish the interferon beta bioavailability in MS patients. Neurology 2003; 60: 634 – 639 .

13.

13 Rudick RA Simonian NA Alam JA . Incidence and significance of neutralizing antibodies to interferon beta-1a in multiple sclerosis. Multiple Sclerosis Collaborative Research Group (MSCRG) [see comments]. Neurology 1998; 50: 1266 – 1272 .

14.

14 Giovannoni G Munschauer F Deisenhammer F . Neutralising antibodies to interferon beta during treatment of multiple sclerosis. J Neurol Neurosurg Psychiatry 2002; 73: 465 – 469 .

15.

15 Ross C Clemmesen KM Sorensen PS Koch-Henriksen N Bendtzen K . Measuring and evaluating interferon beta-induced antibodies in patients with multiple sclerosis. Mult Scler 2006; 12: 39 – 46 .

16.

16 Gneiss C Tripp P Reichartseder F . Differing immunogenic potentials of interferon beta preparations in multiple sclerosis patients. Mult Scler 2006; 12: 731 – 737 .

17.

17 Hesse D Sorensen PS . Using measurements of neutralizing antibodies: the challenge of IFNb therapy. Eur J Neurol 2007; 14: 850 – 859 .

18.

18 Kawade Y . An analysis of neutralization reaction of interferon by antibody: a proposal on the expression of neutralization titer. J Interferon Res 1980; 1: 61 – 70 .

19.

19 Gilli F Hoffmann F Sala A . Qualitative and quantitative analysis of antibody response against IFNbeta in patients with multiple sclerosis. Mult Scler 2006; 12: 738 – 746 .

20.

20 Pachner AR . An improved ELISA for screening for neutralizing anti-IFNb antibodies in MS patients. Neurology 2003; 61: 1444 – 1446 .

21.

21 Petkau AJ White RA Ebers GC . Longitudinal analyses of the effects of neutralizing antibodies on interferon beta-1b in relapsing–remitting multiple sclerosis. Mult Scler 2004; 10: 126 – 138 .

22.

22 Gilli F Marnetto F Caldano M . Anti-interferon-beta neutralising activity is not entirely mediated by antibodies. J Neuroimmunol 2007; 192: 198 – 205 .

23.

23 Deisenhammer F Reindl M Berger T . Immunoglobulin subclasses in patients with serum-binding and neutralizing antibodies against interferon-1b. J Interferon Cytokine Res 2001; 21: 167 – 171 .

24.

24 Gneiss C Reindl M Berger T . Epitope specificity of neutralizing antibodies against IFNb. J Interferon Cytokine Res 2004; 24: 283 – 290 .

25.

25 Gneiss C Tripp P Ehling R . Interferon-beta antibodies have a higher affinity in patients with neutralizing antibodies compared to patients with non-neutralizing antibodies. J Neuroimmunol 2006; 174: 174 – 179 .

26.

26 Lawrence N Oger J Aziz T Palace J Vincent A . A sensitive radioimmunoprecipitation assay for assessing the clinical relevance of antibodies to IFN beta. J Neurol Neurosurg Psychiatry 2003; 74: 1236 – 1239 .

27.

27 Schwab C Bosshard HR . Caveats for the use of surface-adsorbed protein antigen to test the specificity of antibodies. J Immunol Methods 1992; 147: 125 – 134 .