title: Reduction of spiked porcine circovirus during the manufacture of a Vero cell-derived vaccine.
abstract: Porcine circovirus-1 (PCV1) was recently identified as a contaminant in live Rotavirus vaccines, which was likely caused by contaminated porcine trypsin. The event triggered the development of new regulatory guidance on the use of porcine trypsin which shall ensure that cell lines and porcine trypsin in use are free from PCV1. In addition, manufacturing processes of biologicals other than live vaccines include virus clearance steps that may prevent and mitigate any potential virus contamination of product. In this work, artificial spiking of down-scaled models for the manufacturing process of an inactivated pandemic influenza virus vaccine were used to investigate inactivation of PCV1 and the physico-chemically related porcine parvovirus (PPV) by formalin and ultraviolet-C (UV-C) treatment as well as removal by the purification step sucrose gradient ultracentrifugation. A PCV1 infectivity assay, using a real-time PCR infectivity readout was established. The formalin treatment (0.05% for 48h) showed substantial inactivation for both PCV1 and PPV with reduction factors of 3.0log10 and 6.8log10, respectively, whereas UV-C treatment resulted in complete PPV (≥5.9log10) inactivation already at a dose of 13mJ/cm but merely 1.7log10 at 24mJ/cm(2) for PCV1. The UV-C inactivation results with PPV were confirmed using minute virus of mice (MVM), indicating that parvoviruses are far more sensitive to UV-C than PCV1. The sucrose density gradient ultracentrifugation also contributed to PCV1 clearance with a reduction factor of 2log10. The low pH treatment during the production of procine trypsin was investigated and showed effective inactivation for both PCV1 (4.5log10) and PPV (6.4log10). In conclusion, PCV1 in general appears to be more resistant to virus inactivation than PPV. Still, the inactivated pandemic influenza vaccine manufacturing process provides for robust virus reduction, in addition to the already implemented testing for PCV1 to avoid any contaminations.
authors.affiliation: Global Pathogen Safety, Baxter BioScience, Vienna, Austria. Electronic address: cornelia_lackner@baxter.com.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Lackner
authors.affiliation: Global Pathogen Safety, Baxter BioScience, Vienna, Austria. Electronic address: sandra_leydold@baxter.com.
authors.forename: Sandra M
authors.initials: SM
authors.lastname: Leydold
authors.affiliation: Global Pathogen Safety, Baxter BioScience, Vienna, Austria. Electronic address: jens_modrof@baxter.com.
authors.forename: Jens
authors.initials: J
authors.lastname: Modrof
authors.affiliation: Global Pathogen Safety, Baxter BioScience, Vienna, Austria. Electronic address: maria_farcet@baxter.com.
authors.forename: Maria R
authors.initials: MR
authors.lastname: Farcet
authors.affiliation: Cell Culture Fermentation, Baxter BioScience, Orth/Donau, Austria. Electronic address: leopold_grillberger@baxter.com.
authors.forename: Leopold
authors.initials: L
authors.lastname: Grillberger
authors.affiliation: Vaccine R&D, Baxter BioScience, Orth/Donau, Austria. Electronic address: birgit_schaefer@baxter.com.
authors.forename: Birgit
authors.initials: B
authors.lastname: Schäfer
authors.affiliation: Biologicals R&D, Baxter BioScience, Vienna, Austria. Electronic address: heinz_anderle@baxter.com.
authors.forename: Heinz
authors.initials: H
authors.lastname: Anderle
authors.affiliation: Global Pathogen Safety, Baxter BioScience, Vienna, Austria. Electronic address: thomas_kreil@baxter.com.
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Kreil
mesh_terms: D000143:Acids; D000818:Animals; D002522:Chlorocebus aethiops; D017925:Circovirus; D004340:Drug Contamination / Q000517:prevention & control*; D005557:Formaldehyde; D006863:Hydrogen-Ion Concentration; D028323:Parvovirus, Porcine; D013552:Swine; D014461:Ultracentrifugation; D014466:Ultraviolet Rays; D014709:Vero Cells; D014765:Viral Vaccines / Q000494:pharmacology*; D038301:Virus Inactivation
chemical_list: D000143:Acids; D014765:Viral Vaccines; D005557:Formaldehyde
keywords: Adventitious virus; Influenza; Porcine circovirus; Porcine parvovirus; Trypsin; Vaccine; Vero-cell platform