Modeling rotavirus-like particles production in a baculovirus expression vector system : Infection kinetics, baculovirus DNA replication, mRNA synthesis and protein production - CNRS - Centre national de la recherche scientifique Accéder directement au contenu
Article Dans Une Revue Journal of Biotechnology Année : 2007

Modeling rotavirus-like particles production in a baculovirus expression vector system : Infection kinetics, baculovirus DNA replication, mRNA synthesis and protein production

Résumé

Rotavirus is the most common cause of severe diarrhoea in children worldwide, responsible for more than half a million deaths in children per year. Rotavirus-like particles (Rota VLPs) are excellent vaccine candidates against rotavirus infection, since they are non-infectious, highly immunogenic, amenable to large-scale production and safer to produce than those based on attenuated viruses. This work focuses on the analysis and modeling of the major events taking place inside Spodoptera frugiperda (Sf-9) cells infected by recombinant baculovirus that may be critical for the expression of rotavirus viral proteins (VPs). For model validation, experiments were performed adopting either a co-infection strategy, using three monocistronic recombinant baculovirus each one coding for viral proteins VP2, VP6 and VP7, or single-infection strategies using a multigene baculovirus coding for the three proteins of interest. A characteristic viral DNA (vDNA) replication rate of 0.19 +/- 0.01 h(-1) was obtained irrespective of the monocistronic or multigene vector employed, and synthesis of progeny virus was found to be negligible in comparison to intracellular vDNA concentrations. The timeframe for vDNA, mRNA and VP synthesis tends to decrease with increasing multiplicity of infection (MOI) due to the metabolic burden effect. The protein synthesis rates could be ranked according to the gene size in the multigene experiments but not in the co-infection experiments. The model exhibits acceptable prediction power of the dynamics of intracellular vDNA replication, mRNA synthesis and VP production for the three proteins involved. This model is intended to be the basis for future Rota VLPs process optimisation and also a means to evaluating different baculovirus constructs for Rota VLPs production.

Dates et versions

hal-02663708 , version 1 (31-05-2020)

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Citer

Antonio Roldão, Helena L.A. Vieira, Annie Charpilienne, Didier Poncet, Polly Roy, et al.. Modeling rotavirus-like particles production in a baculovirus expression vector system : Infection kinetics, baculovirus DNA replication, mRNA synthesis and protein production. Journal of Biotechnology, 2007, 128 (4), pp.875-894. ⟨10.1016/j.jbiotec.2007.01.003⟩. ⟨hal-02663708⟩
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