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Clinically important neutralizing anti-drug antibodies detected with an in-house competitive ELISA

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Abstract

Therapeutic drug monitoring of TNF-alpha inhibitors is crucial for evaluating patients with inflammatory diseases on a personalized level. It has been clinically observed that many patients receiving TNF-alpha inhibitors, with negative drug and anti-drug antibody results from bridging ELISA (bELISA), lose their drug response over time, despite dose optimization. Our aims were to develop innovative in-house competitive ELISAs (cELISAs) for the detection of neutralizing antibodies against infliximab and adalimumab and compare their results to reporter gene assay (RGA) and in-house bELISA. Furthermore, we aimed to evaluate patient anti-drug antibody results in regard to their clinical records and potential benefits of therapeutic drug monitoring with the novel cELISAs. Sera of patients treated with infliximab (n = 46) or adalimumab (n = 31), having undetectable drug levels, were tested with our in-house cELISA. Briefly, samples were incubated with a fixed amount of drug and the neutralizing capacity of the samples was determined. The cELISA results were compared to RGA and bELISA results using Spearman’s correlation coefficient. Additionally, patient clinical data were evaluated in line with the results of cELISA, bELISA, and RGA using the Kaplan-Meier analysis and the Log Rank test. Both anti-infliximab and anti-adalimumab cELISAs showed very good correlation to RGA (r = 0.932, p < 0.0001 and r = 0.947, p < 0.0001, respectively). Furthermore, a positive result in anti-infliximab cELISA can predict treatment failure in 100% of patients with negative bELISA, while a positive result in anti-adalimumab cELISA can predict treatment failure in 80% of patients with negative bELISA. Taken together, we developed innovative cELISAs enabling quantification of functional and neutralizing anti-drug antibodies, comparable to RGA. The association between cELISA results and loss of drug response in patients identified clinically important anti-drug antibodies, as measured by cELISA.

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Acknowledgments

The authors would like to thank all the staff in the Department of Rheumatology and the Immunology Laboratory, who helped collect and process the patient samples.

Funding

The Slovenian Research Agency funded the National Research Program “Systemic Autoimmune Diseases” #P3-0314.

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Authors and Affiliations

  1. Department of Rheumatology, University Medical Centre Ljubljana, SI-1000, Ljubljana, Slovenia

    Manca Ogrič, Polona Žigon, Katja Lakota, Sonja Praprotnik, Snezna Sodin-Semrl & Saša Čučnik

  2. Faculty of Mathematics, Natural Sciences and Information Technologies, University of Primorska, SI-6000, Koper, Slovenia

    Katja Lakota & Snezna Sodin-Semrl

  3. Department of Gastroenterology, University Medical Centre Ljubljana, SI-1000, Ljubljana, Slovenia

    David Drobne & Borut Štabuc

  4. Faculty of Medicine, University of Ljubljana, SI-1000, Ljubljana, Slovenia

    David Drobne & Borut Štabuc

  5. Faculty of Pharmacy, Chair of Clinical Biochemistry, University of Ljubljana, SI-1000, Ljubljana, Slovenia

    Saša Čučnik

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  1. Manca Ogrič
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  7. Snezna Sodin-Semrl
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  8. Saša Čučnik
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All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

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Correspondence to Saša Čučnik.

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Ethical approval

This study was conducted as part of the National Research Program (#P3-0314) and has been approved by the National Medical Ethics Committee (#99/04/15 and #38/01/16). It has therefore been performed in accordance with the ethical standards laid down in 1964 Declaration of Helsinki and its later amendments. Informed consent was obtained from all individuals included in this study.

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No funding commercial organizations played a role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

Additional information

Rheumatology in Slovenia: Clinical practice and translational research

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Ogrič, M., Žigon, P., Lakota, K. et al. Clinically important neutralizing anti-drug antibodies detected with an in-house competitive ELISA. Clin Rheumatol 38, 361–370 (2019). https://doi.org/10.1007/s10067-018-4213-0

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  • DOI: https://doi.org/10.1007/s10067-018-4213-0

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