Large scale production and downstream processing of a recombinant porcine parvovirus vaccine

L. Maranga, P. Rueda, A.F.G. Antonis, C. Vela, J.P.M. Langeveld, J.I. Casal, M.J.T. Carrondo

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    48 Citations (Scopus)

    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 1.52106 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.
    Original languageEnglish
    Pages (from-to)45-50
    JournalApplied Microbiology and Biotechnology
    Volume59
    DOIs
    Publication statusPublished - 2002

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