Sf9 Cells Metabolism and Viability When Coinfected with Two Monocistronic Baculoviruses to Produce Rabies Virus-like Particles

Autor(es)

Leme, Jaci ; Oliveira Guardalini, Luis Giovani ; Bernardino, Thaissa Consoni ; Astray, Renato Mancini ; Tonso, Aldo ; Fernandez Nunez, Eutimio Gustavo ; Calil Jorge, Soraia Attie

• Tipo de documento: Artigo Científico
• Fonte: MOLECULAR BIOTECHNOLOGY; v. N/A, p. 13-pg., 2022-11-18.

Resumo

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 different multiplicities of infection (MOI). Schott flasks 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 glutamate 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 specific death rate depend on BV MOI. The wave pattern of nutrient/metabolite profiles throughout the viral infection phase is related to the baculo-virus lytic cycle. The optimal MOIs ratio between BVG (2.5-4.5) and BVM (1.0-3.0) for maximum protein expression was defined. 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 specifically, for progressing in a rabies VLP vaccine development. [GRAPHICS] . (AU)

• Processo FAPESP: 16/22780-6 - Produção e caracterização de VLPs de Arbovírus produzidos em sistemas de células de inseto.
• Beneficiário: Soraia Attie Calil Jorge
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 18/10538-1 - Caracterização de antígenos recombinantes e da resposta imune para um novo candidato vacinal contra a raiva
• Beneficiário: Renato Mancini Astray
• Modalidade de apoio: Auxílio à Pesquisa - Regular