title: Large scale production and downstream processing of a recombinant porcine parvovirus vaccine.
abstract: Porcine parvovirus (PPV) virus-like particles (VLPs) constitute a potential vaccine for prevention of parvovirus-induced reproductive failure in gilts. Here we report the development of a large scale (25 l) production process for PPV-VLPs with baculovirus-infected insect cells. A low multiplicity of infection (MOI) strategy was efficiently applied avoiding the use of an extra baculovirus expansion step. The optimal harvest time was defined at 120 h post-infection at the MOI used, with the cell concentration at infection being 1.5x10(6) cells/ml. An efficient purification scheme using centrifugation, precipitation and ultrafiltration/diafiltration as stepwise unit operations was developed. The global yield of the downstream process was 68%. Baculovirus inactivation with Triton X-100 was successfully integrated into the purification scheme without an increase in the number of process stages. Immunogenicity of the PPV-VLPs tested in guinea pigs was similar to highly purified reference material produced from cells cultured in the presence of serum-containing medium. These results indicate the feasibility of industrial scale production of PPV-VLPs in the baculovirus system, safety of the product, and the potency of the product for vaccine application.
authors.affiliation: Instituto de Biologia Experimental e Tecnológica/Instituto de Tecnologia Química e Biológica IBET/ITQB, Apartado 12, 2781-901 Oeiras, Portugal.
authors.forename: L
authors.initials: L
authors.lastname: Maranga
authors.forename: P
authors.initials: P
authors.lastname: Rueda
authors.forename: A F G
authors.initials: AF
authors.lastname: Antonis
authors.forename: C
authors.initials: C
authors.lastname: Vela
authors.forename: J P M
authors.initials: JP
authors.lastname: Langeveld
authors.forename: J I
authors.initials: JI
authors.lastname: Casal
authors.forename: M J T
authors.initials: MJ
authors.lastname: Carrondo
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis; D016367:Baculoviridae / Q000235:genetics; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D005240:Feasibility Studies; D006168:Guinea Pigs; D007115:Immunization Schedule; D028323:Parvovirus, Porcine / Q000276:immunology*; D018411:Spodoptera / Q000166:cytology; D013678:Technology, Pharmaceutical; D014611:Vaccination / Q000662:veterinary; D014614:Vaccines, Synthetic; D014765:Viral Vaccines / Q000276:immunology; D014771:Virion / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D016895:Culture Media, Serum-Free; D014614:Vaccines, Synthetic; D014765:Viral Vaccines