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Quartz crystal microbalance as an assay to detect anti-drug antibodies for the immunogenicity assessment of therapeutic biologics

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Abstract

Because of their biological origins, therapeutic biologics can trigger an unwanted deleterious immune response with some patients. The immunogenicity of therapeutic biologics can affect drug efficacy and patient safety by the production of circulating anti-drug antibodies (ADA). In this study, quartz crystal microbalance (QCM) was developed as an assay to detect ADA. Etanercept (Enbrel®) was covalently grafted to dextran-modified QCM surfaces. Rabbits were immunized with etanercept to generate ADA. Results showed the QCM assay could detect purified ADA from rabbits at concentrations as low as 50 ng/mL, within the sensitivity range of ELISA. The QCM assay could also assess the ADA isotype. It was shown that the ADA were composed of the IgG isotype, but not IgM, as expected. Furthermore, it was shown that QCM surfaces that had been used to detect ADA could be regenerated in glycine-HCl solution and reused. The QCM assay was also demonstrated to detect ADA in crude serum samples. Serum was collected from the rabbits and analyzed before and after etanercept immunization. ADA were clearly detected in serum from rabbits after immunization, but not in serum before immunization. Serum from patients administered with etanercept for rheumatoid arthritis (RA) treatment was also analyzed and compared to serum from healthy donors. Sera from 10 RA patients were analyzed. Results showed one of the RA patient serum samples may have ADA present. In conclusion, QCM appears to be a viable assay to detect ADA for the immunogenicity assessment of therapeutic biologics.

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Acknowledgements

We would like to acknowledge Dr. Brian Granda for his advice.

Funding

The authors are grateful to the Québec Consortium for Drug Discovery (CQDM, Consortium québécois sur la découverte du médicament) for their financial support of this work.

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Authors and Affiliations

  1. Laboratoire de bio-ingénierie et de biophysique de l’Université de Sherbrooke, Department of Chemical and Biotechnological Engineering, Université de Sherbrooke, 2500 boulevard de l’Université, Sherbrooke, Québec, J1K 2R1, Canada

    Evan A. Dubiel, Sylvain Vigier & Patrick Vermette

  2. Research Centre on Aging, Institut universitaire de gériatrie de Sherbrooke, 1036 rue Belvédère Sud, Sherbrooke, Québec, J1H 4C4, Canada

    Evan A. Dubiel, Tamás Fülöp, Sylvain Vigier & Patrick Vermette

  3. Faculté de médecine et des sciences de la santé, Institut de pharmacologie de Sherbrooke, 3001 12ième Avenue Nord, Sherbrooke, Québec, J1H 5N4, Canada

    Evan A. Dubiel & Patrick Vermette

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  1. Evan A. Dubiel
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  2. Tamás Fülöp
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Correspondence to Patrick Vermette.

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The collection of all serum samples was done in accordance with approval of the Ethical Committee of the Université de Sherbrooke (Dossier # 2015-490). Work with rabbits was performed in accordance with internal ethical guidelines of Biomatik.

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Dubiel, E.A., Fülöp, T., Vigier, S. et al. Quartz crystal microbalance as an assay to detect anti-drug antibodies for the immunogenicity assessment of therapeutic biologics. Anal Bioanal Chem 409, 7153–7167 (2017). https://doi.org/10.1007/s00216-017-0674-2

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