title: The Protein/Peptide Direct Virus Inactivation During Chromatographic Process: Developing Approaches.
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.
authors.affiliation: Neutromics Ukraina TOV, 12, Melnikova str, Kyiv, Ukraine. volkov.georgii@neutromics.com.ua.|Shijir International LCC, Raining Valley, Ulaanbaatar, Mongolia. volkov.georgii@neutromics.com.ua.
authors.forename: Georgii L
authors.initials: GL
authors.lastname: Volkov
authors.affiliation: SPC Interpharmbiotech TOV, 150, Zabolotnogo str, Kyiv, 03680, Ukraine.
authors.forename: Sergiy P
authors.initials: SP
authors.lastname: Havryliuk
authors.affiliation: Covance Clinical and Periapproval Services LLC, 4, Hrinchenka str, Kyiv, Ukraine.
authors.forename: Ievgenia M
authors.initials: IM
authors.lastname: Krasnobryzha
authors.affiliation: Neutromics Ukraina TOV, 12, Melnikova str, Kyiv, Ukraine.|Shijir International LCC, Raining Valley, Ulaanbaatar, Mongolia.
authors.forename: Olena S
authors.initials: OS
authors.lastname: Havryliuk
mesh_terms: D000818:Animals; D006801:Humans; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000627:therapeutic use; D012997:Solvents / Q000737:chemistry; D014777:Virus Diseases / Q000188:drug therapy* / Q000821:virology; D038301:Virus Inactivation / Q000187:drug effects*; D014780:Viruses / Q000187:drug effects*
chemical_list: D010455:Peptides; D011994:Recombinant Proteins; D012997:Solvents
keywords: Chromatography; Dynamic capacity; Infectivity; Peptide; Protien; RT-PCR; Virus inactivation; Virus titer