title: Large scale production of a mammalian cell derived quadrivalent hepatitis C virus like particle vaccine.
abstract: A method for the large-scale production of a quadrivalent mammalian cell derived hepatitis C virus-like particles (HCV VLPs) is described. The HCV core E1 and E2 coding sequences of genotype 1a, 1b, 2a or 3a were co-expressed in Huh7 cell factories using a recombinant adenoviral expression system. The structural proteins self-assembled into VLPs that were purified from Huh7 cell lysates by iodixanol ultracentrifugation and Stirred cell ultrafiltration. Electron microscopy, revealed VLPs of the different genotypes that are morphologically similar. Our results show that it is possible to produce large quantities of individual HCV genotype VLPs with relative ease thus making this approach an alternative for the manufacture of a quadrivalent mammalian cell derived HCV VLP vaccine.
authors.affiliation: Department of Microbiology and Immunology, The Peter Doherty Institute for Infection and Immunity, University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: L
authors.initials: L
authors.lastname: Earnest-Silveira
authors.affiliation: Department of Microbiology and Immunology, The Peter Doherty Institute for Infection and Immunity, University of Melbourne, Parkville, Victoria 3010, Australia; Department of Surgery, Austin Hospital, University of Melbourne, Australia.
authors.forename: D
authors.initials: D
authors.lastname: Christiansen
authors.affiliation: Department of Biochemistry and Molecular Biology, Bio21 Institute of Molecular Science and Biotechnology, The University of Melbourne, Australia.
authors.forename: S
authors.initials: S
authors.lastname: Herrmann
authors.affiliation: Department of Biochemistry and Molecular Biology, Bio21 Institute of Molecular Science and Biotechnology, The University of Melbourne, Australia.
authors.forename: S A
authors.initials: SA
authors.lastname: Ralph
authors.affiliation: Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore, Karnataka 560012, India.
authors.forename: S
authors.initials: S
authors.lastname: Das
authors.affiliation: The Basil Hetzel Institute and Queen Elizabeth Hospital, University of Adelaide, South Australia, Australia.
authors.forename: E J
authors.initials: EJ
authors.lastname: Gowans
authors.affiliation: Department of Microbiology and Immunology, The Peter Doherty Institute for Infection and Immunity, University of Melbourne, Parkville, Victoria 3010, Australia. Electronic address: josepht@unimelb.edu.au.
authors.forename: J
authors.initials: J
authors.lastname: Torresi
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D002460:Cell Line; D015870:Gene Expression; D005822:Genetic Vectors; D016174:Hepacivirus / Q000235:genetics*; D022781:Hepatocytes / Q000378:metabolism / Q000821:virology; D006801:Humans; D046529:Microscopy, Electron, Transmission; D058425:Vaccines, Virus-Like Particle / Q000302:isolation & purification* / Q000648:ultrastructure; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D058425:Vaccines, Virus-Like Particle; D014764:Viral Proteins
keywords: Hepatitis C; Quadrivalent vaccine; Virus-like particles