Sf9 Cells metabolism and viability when coinfected with two monocistronic baculoviruses to produce rabies virus‑like particles
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This work aimed to describe the dynamics of the Sf9 insect cells death and primary metabolism when this host is infected simultaneously by two recombinant baculoviruses (BV) expressing rabies glycoprotein (BVG) and matrix protein (BVM) genes to produce rabies virus-like particles (VLP) at diferent multiplicities of infection (MOI). Schott fasks essays covering a wide range of MOI for both BV were performed. Viable cell density, cell viability, glucose, glutamine, glutamate, lactate, ammonium, and rabies proteins concentrations were monitored over the infection phase. The expression of both recombinant proteins was not limited by glucose, glutamine, and lutamate in a broad MOI (pfu/cell) range of BVG (0.15–12.5) and BVM (0.1–5.0) using SF900 III serum free culture medium. Death phase initiation and the specifc death rate depend on BV MOI. The wave pattern of nutrient/metabolite profles throughout the viral infection phase is related to the baculovirus lytic cycle. The optimal MOIs ratio between BVG (2.5–4.5) and BVM (1.0–3.0) for maximum protein expression was defned. The produced rabies VLP sizes are close to 78 nm. In general, these work outputs bring a better understanding of the metabolic performance of Sf9 cells when infected by BV for producing VLP, and specifcally, for progressing in a rabies VLP vaccine development.
Leme J, Guardalini LGO, Bernardino TC, Astray RM, Tonso A, Núñez EGF, et al. Sf9 Cells Metabolism and Viability When Coinfected with Two Monocistronic Baculoviruses to Produce Rabies Virus‑like Particles. Mol Biotechnol. 2022 Nov; 65:970–982. doi:10.1007/s12033-022-00586-x.
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