Large scale production and downstream processing of a recombinant porcine parvovirus vaccine
| Autor: | Adriaan F.G. Antonis, Paloma Rueda, J.I. Casal, Manuel J.T. Carrondo, Luis Maranga, C. Vela, Jan P. M. Langeveld |
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| Jazyk: | angličtina |
| Rok vydání: | 2002 |
| Předmět: |
Porcine parvovirus
viruses Guinea Pigs Spodoptera Antibodies Viral Applied Microbiology and Biotechnology Culture Media Serum-Free Microbiology Multiplicity of infection Virus-like particle Animals Technology Pharmaceutical Life Science Instituut voor Dierhouderij en Diergezondheid Cells Cultured Immunization Schedule Vaccines Synthetic Downstream processing biology ID-Lelystad Parvovirus Immunogenicity Viral Vaccine Vaccination Virion Viral Vaccines General Medicine Parvovirus Porcine biology.organism_classification Virology ID Lelystad Diafiltration ID-Lelystad Instituut voor Dierhouderij en Diergezondheid ID Lelystad Institute for Animal Science and Health Feasibility Studies Baculoviridae Institute for Animal Science and Health Biotechnology |
| Zdroj: | Applied Microbiology and Biotechnology 59 (2002) Digital.CSIC. Repositorio Institucional del CSIC instname Applied Microbiology and Biotechnology, 59, 45-50 |
| ISSN: | 0175-7598 |
| Popis: | 6 páginas, 6 figuras, 1 tabla -- PAGS nros. 45-50 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 1.5×106 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 The authors acknowledge and appreciate the financial support received for the project from the European Commission (BIO4-CT98-0215) and a grant to L.M. from Fundação para a Ciência e Tecnologia – Portugal (PRAXIS XXI/BD/ 16136/98) |
| Databáze: | OpenAIRE |
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