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Measuring the bioactivity of anti-IL-6/anti-IL-6R therapeutic antibodies: presentation of a robust reporter gene assay

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Abstract

IL-6 has an important role in the pathogenesis of autoimmunity and chronic inflammation. Several mAbs that target IL-6 or the IL-6 receptor (IL-6R) have been established and approved for the treatment of various diseases such as multicentric Castleman's disease and rheumatoid arthritis. Quality control of therapeutic antibodies requires accurate determination of bioactivity. However, current cell-based anti-proliferation assays are tedious, time consuming, and result in high variation. We therefore developed a reporter gene assay (RGA) based on an IL-6-dependent DS-1 cell line that stably expressed the reporter luciferase controlled by the serum-induced element (SIE) response element, which was a key element located downstream of the IL-6 signaling pathway. The RGA method demonstrated good performance characteristics after careful optimization, including high specificity, stability, accuracy, precision, and robustness. It also had superior precision and sensitivity. The assay is simple compared with the traditional anti-proliferation assay. This novel RGA based on the IL-6-IL-6R-STAT3 pathway can be useful, in conjunction with the anti-proliferation bioassay, to determine the bioactivity of anti-IL-6/anti-IL-6R therapeutic mAbs.

The mechanism sketch of the reporter gene assay for the bioactivity determination of anti-IL-6/anti-IL-6Rα mAbs

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Acknowledgments

This work was supported by Research of Evaluation of Innovative Biopharmaceuticals and Standardization Technology (one of the subjects of the National Science and Technology Major Projects for ‘Major New Drugs Innovation and Development’) under grant No. 2018ZX09101001.

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Author notes

  1. Chuanfei Yu, Junxia Cao, and Lan Wang contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products, National Institutes for Food and Drug Control, No. 2, Tiantan Xili, Dongcheng District, Beijing, 100050, China

    Chuanfei Yu, Junxia Cao, Lan Wang, Yalan Yang, Yongbo Ni & Junzhi Wang

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  1. Chuanfei Yu
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  2. Junxia Cao
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  3. Lan Wang
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Correspondence to Junzhi Wang.

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Yu, C., Cao, J., Wang, L. et al. Measuring the bioactivity of anti-IL-6/anti-IL-6R therapeutic antibodies: presentation of a robust reporter gene assay. Anal Bioanal Chem 410, 7067–7075 (2018). https://doi.org/10.1007/s00216-018-1307-0

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  • DOI: https://doi.org/10.1007/s00216-018-1307-0

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