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The Protein/Peptide Direct Virus Inactivation During Chromatographic Process: Developing Approaches

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Abstract

Virus clearance is required for pharmaceutical preparations derived from animal or human sources such as blood products, vaccines, recombinant proteins produced in mammalian cell lines, etc. High cost and substantial protein losses during virus inactivation are significant problems for protein/peptide manufacturing. The goal of this project was to develop a method to perform virus inactivation in a course of protein chromatographic purification. Another goal was to show that the chromatographic adsorbent can serve as reliable “sieva” for mechanical washing away of infecting viruses. Using chromatographic, photometric, IFA, and RT-PCR approaches, it was discovered that high temperature-depending dynamic capacity of adsorbent allowed to perform a virus inactivation directly in a chromatographic column by solvent/detergent treatment. The peptide/protein biological activity was completely preserved. Using this new approach enveloped and nonenveloped viruses were effectively removed protein preparation. In addition, it was shown that RT-PCR method demonstrates more precise and reproducible results and robust properties for assessment of virus reduction than virus titer followed by infectivity studies. Presented method allowed to obtain the factor of virus concentration decrease (FVD) values that were higher than those provided by known technologies and was sufficient for a full inactivation of viruses. The method is recommended to use in pharmaceutical industry.

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Acknowledgments

The authors express profound gratitude to the staff of pilot plant of scientific and manufacturing firm Shijir International Co. Ltd., Sukhbaatar sq., Bodi Tower building, Ulaanbaatar, Mongolia for the opportunity to intervene in the real manufacturing processes and to test the represented method on the pilot plant in the Raining (Boroo) Valley of Mongolia.

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Authors and Affiliations

  1. Neutromics Ukraina TOV, 12, Melnikova str, Kyiv, Ukraine

    Georgii L. Volkov & Olena S. Havryliuk

  2. Shijir International LCC, Raining Valley, Ulaanbaatar, Mongolia

    Georgii L. Volkov & Olena S. Havryliuk

  3. SPC Interpharmbiotech TOV, 150, Zabolotnogo str, Kyiv, 03680, Ukraine

    Sergiy P. Havryliuk

  4. Covance Clinical and Periapproval Services LLC, 4, Hrinchenka str, Kyiv, Ukraine

    Ievgenia M. Krasnobryzha

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  1. Georgii L. Volkov
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  2. Sergiy P. Havryliuk
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  3. Ievgenia M. Krasnobryzha
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  4. Olena S. Havryliuk
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Correspondence to Georgii L. Volkov.

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Volkov, G.L., Havryliuk, S.P., Krasnobryzha, I.M. et al. The Protein/Peptide Direct Virus Inactivation During Chromatographic Process: Developing Approaches. Appl Biochem Biotechnol 181, 233–249 (2017). https://doi.org/10.1007/s12010-016-2209-2

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