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Assessment of biosimilarity under native and heat-stressed conditions: rituximab, bevacizumab, and trastuzumab originators and biosimilars

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Abstract

Biosimilars are highly similar to, but not identical with, their originator products. As a result, structural differences between originators and biosimilars can be difficult to detect and characterize without the appropriate analytical tools. Therefore, we first focus on identifying initial structural differences between rituximab, bevacizumab, and trastuzumab originator and biosimilar pairs and later address how these differences change after applying thermal stress at 40 °C with orbital shaking for 4 weeks. Prior to incubation, we detected comparable secondary and tertiary structures for each pair and identified different levels of soluble aggregates, charge variants, and molecular weight variants due to differences in glycoforms and the number of C-terminal lysine groups. Over the course of incubation, we compared differences in charge variants and unfolding patterns. Taken together, our study provides a comparability exercise, providing information on the minor differences present between originator and biosimilar products and how those differences are impacted by stress.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Michigan, Ann Arbor, MI, USA

    Jukyung Kang, Zeynab Izadi Najafabadi & Anna Schwendeman

  2. Biointerfaces Institute, University of Michigan, 2800 Plymouth Road, Ann Arbor, MI, 48109, USA

    Jukyung Kang, Troy Halseth & Anna Schwendeman

  3. Department of Medicinal Chemistry, University of Michigan, Ann Arbor, MI, USA

    Troy Halseth & Anna Schwendeman

  4. Department of Chemistry, University of Michigan, Ann Arbor, MI, USA

    Daniel Vallejo & Brandon T. Ruotolo

  5. Protein Metrics Inc., San Carlos, CA, USA

    K. Ilker Sen

  6. MS Bioworks, Ann Arbor, MI, USA

    Michael Ford

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Kang, J., Halseth, T., Vallejo, D. et al. Assessment of biosimilarity under native and heat-stressed conditions: rituximab, bevacizumab, and trastuzumab originators and biosimilars. Anal Bioanal Chem 412, 763–775 (2020). https://doi.org/10.1007/s00216-019-02298-9

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