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Comparative analytical profiling of bevacizumab biosimilars marketed in India: a national control laboratory study

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Abstract

In this study, analytical profiling of the bevacizumab (BVZ) biosimilars (N = 3) approved in India were evaluated for charge heterogeneity, isoelectric focusing, aggregation and in vitro potency analysis. The charge variants were characterized using high performance cation-exchange chromatography (CEX-HPLC), capillary zone electrophoresis (CZE) and capillary isoelectric focusing (cIEF). cIEF was also used for estimation of isoelectric point (pI value). In addition, aggregate analysis was done using size exclusion high performance chromatography (SEC-HPLC). The cell-based inhibition of proliferation assay using HUVEC cells, indirect ELISA and Western blot were performed for in vitro biological activity. In addition of cell-based cytotoxicity assay was also performed and found no cytotoxic effect on both HuT78 and WIL2S cells by bevacizumab biosimilars. The significant variations in acidic (p < 0.0001) and basic variants (p < 0.0001), pI value (p = 0.0035), aggregates (p = 0.0306) of biosimilars were found as compared to innovator product; however, cell-based potency analysis (p = 0.6047) and indirect ELISA (p = 0.1611) have shown no significant difference in the biological activity. The banding patterns of all biosimilars in western blot were found similar to the innovator product. The comparatively higher basic variants in the biosimilars were attributing to the high pI value of biosimilars to that of innovator product, although these variations were not affecting the biological activity of the biosimiars. This is a unique study, wherein the independent analysis by a National Control Laboratory (NCL) will not only help the National Regulatory Authority (NRA) to assess the quality and consistency in manufacturing of BVZ biosimilars marketed in India but also facilitate the uptake of BVZ biosimilars, and sustainable access to new medicines against the anti-angiogenic therapy.

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Acknowledgements

Our study team is thankful to CDSCO for providing the bevacizumab samples for quality evaluation. Study was fully funded by National Institute of Biologicals (Ministry of Health and Family Welfare), (Grant no. NIB-TAL) Government of India.

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Author notes

  1. Utpreksha Vaish

    Present address: Miller School of Medicine, University of Miami, Florida, USA

  2. Anu Prakash and Nripendra N. Mishra have equal first author.

Authors and Affiliations

  1. National Institute of Biologicals, (Ministry of Health and Family Welfare, Govt. of India), A-32, Sector 62, Noida, 201309, India

    Anu Prakash, Nripendra N. Mishra, Utpreksha Vaish, Sonia Sharma, Apoorva Anand, Richi V. Mahajan, J. P. Prasad & Subhash Chand

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  1. Anu Prakash
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Correspondence to Subhash Chand.

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Prakash, A., Mishra, N.N., Vaish, U. et al. Comparative analytical profiling of bevacizumab biosimilars marketed in India: a national control laboratory study. 3 Biotech 10, 516 (2020). https://doi.org/10.1007/s13205-020-02506-9

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