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Evaluation of Structural, Biological, and Functional Similarity of Biosimilar Granulocyte Colony Stimulating Factor to its Reference Product

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Abstract

Purpose

Granulocyte colony stimulating factor (GCSF; also known as filgrastim) is a growth factor used to induce production of granulocytes. As the first locally developed and approved biosimilar medicine of Turkey, Fraven® being a biosimilar of filgrastim has been ab initio manufactured from cell to finished product at two different production facilities. Comprehensive structural, biological and functional characterization studies were performed to compare Fraven® from two different production sites and its reference product Neupogen® sourced from Turkey.

Methods

Primary and higher-order protein structures were analyzed by high performance liquid chromatography electrospray ionization-time of flight mass spectrometry, circular dichroism, and two-dimensional nuclear magnetic resonance spectroscopy. Isoelectric focusing, SDS-Page, size exclusion chromatography, and related proteins analyses were used to compare impurities. In order to assess functional similarity, surface plasmon resonance (SPR) was used. In vitro cell proliferation assay was also performed to show dose related drug response in NFS-60 cell line.

Results

Primary, secondary and tertiary structures of biosimilar Fraven® manufactured at both sites were found to be highly similar to the reference Neupogen®. Product related substances and impurities were also highly similar to the reference. Comparability of GCSF receptor binding affinities of each product was shown using the KD values of SPR analysis. In vitro cell proliferation similarity was also evaluated and proven by PLA.

Conclusion

Based on the similarity assessment, despite being manufactured at two different production sites, biosimilar Fraven® is highly similar to the reference product Turkey originated Neupogen®.

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Acknowledgments

All analyses were funded by Arven and partially by Scientific and Technological Research Council of Turkey (TUBITAK; 3100431). Study design, management, analysis, and interpretation of analytical data were done by the authors and other Arven personnel. We gratefully acknowledge our former team members who also contributed to the development of the biosimilar filgrastim, Fraven®.

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Author notes

  1. Zeynep Bor Tekdemir and Ali Ihsan Seckin contributed equally to this work.

Authors and Affiliations

  1. Arven Pharmaceutical Research and Development Center, Selimpasa Merkez Mahallesi, 34590, Silivri/Istanbul, Turkey

    Zeynep Bor Tekdemir, Ali Ihsan Seckin, Turgay Kacar & Emine Yilmaz

  2. TUBITAK Marmara Research Center, Institute of Chemical Technology, P.O. Box 21, 41470, Gebze/Kocaeli, Turkey

    Somer Bekiroglu

Authors
  1. Zeynep Bor Tekdemir
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  2. Ali Ihsan Seckin
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  3. Turgay Kacar
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  4. Emine Yilmaz
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  5. Somer Bekiroglu
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Correspondence to Zeynep Bor Tekdemir.

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Bor Tekdemir, Z., Seckin, A.I., Kacar, T. et al. Evaluation of Structural, Biological, and Functional Similarity of Biosimilar Granulocyte Colony Stimulating Factor to its Reference Product. Pharm Res 37, 215 (2020). https://doi.org/10.1007/s11095-020-02932-7

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  • DOI: https://doi.org/10.1007/s11095-020-02932-7

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