Sf9 Cell Metabolism Throughout the Recombinant Baculovirus and Rabies Virus-Like Particles Production in Two Culture Systems

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Guardalini, Luis Giovani Oliveira ; Leme, Jaci ; Cavalcante, Paulo Eduardo da Silva ; de Mello, Renata Gois ; Bernardino, Thaissa Consoni ; Jared, Simone Goncalves Silva ; Antoniazzi, Marta Maria ; Astray, Renato Mancini ; Tonso, Aldo ; Nunez, Eutimio Gustavo Fernandez ; Jorge, Soraia Attie Calil

• Tipo de documento: Artigo Científico
• Fonte: MOLECULAR BIOTECHNOLOGY; v. N/A, p. 11-pg., 2023-05-10.

Resumo

This work aimed to assess the Sf9 cell metabolism during growth, and infection steps with recombinant baculovirus bearing rabies virus proteins, to finally obtain rabies VLP in two culture systems: Schott flask (SF) and stirred tank reactor (STR). Eight assays were performed in SF and STR (four assays in each system) using serum-free SF900 III culture medium. Two non-infection growth kinetics assays and six recombinant baculovirus infection assays. The infection runs were carried out at 0.1 pfu/cell multiplicity of infection (MOI) for single baculovirus bearing rabies glycoprotein (BVG) and matrix protein (BVM) and a coinfection with both baculoviruses at MOI of 3 and 2 pfu/cell for BVG and BVM, respectively. The SF assays were done in triplicate. The glucose, glutamine, glutamate, lactate, and ammonium uptake or release specific rates were quantified over the exponential growth phase and infection stage. The highest uptake specific rate was observed for glucose (42.5 x 10(-12) mmol cell/h) in SF and for glutamine (30.8 x 10(-12) mmol/cell/h) in STR, in the exponential growth phases. A wave pattern was observed for assessed analytes throughout the infection phase and the glucose had the highest wave amplitude within the 10(-10) mmol cell/h order. This alternative uptake and release behavior is in harmony with the lytic cycle of baculovirus in insect cells. The virus propagation and VLP generation were not limited by glucose, glutamine, and glutamate, neither by the toxicity of lactate nor ammonium under the conditions appraised in this work. The findings from this work can be useful to set baculovirus infection processes at high cell density to improve rabies VLP yield, purity, and productivity. (AU)

• 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
• 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