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Bernardino, Thaissa Consoni ; Teruya, Milena Miyu ; Cavalcante, Paulo Eduardo da Silva ; Dias, Vinicius Aragao Tejo ; Rabello, Julia Publio ; Barrence, Fernanda Angela Correia ; Leme, Jaci ; Guardalini, Luis Giovani de Oliveira ; Tonso, Aldo ; Jorge, Soraia Attie Calil ; Nunez, Eutimio Gustavo Fernandez

• Tipo de documento: Artigo Científico
• Fonte: Bioprocess and Biosystems Engineering; v. 48, n. 12, p. 16-pg., 2025-09-22.

Resumo

Zika virus (ZIKV) was declared a public health emergency in 2016, yet effective vaccines are still needed. Among the immunization platforms under evaluation, virus-like particles (VLP) are promising candidates. Growth, metabolism, and respiration are among the cell host processes that are essential for optimizing and characterizing VLP upstream production stage. These cell functions can be influenced by factors such as culture medium composition and the multiplicity of infection (MOI) in viral vector-based expression systems. This study investigated the effects of three MOIs (2, 6, and 10) in a baculovirus/Sf9 insect cell system on ZIKV VLP production with and without medium supplemented with 0.028 mM cholesterol and 6 nM albumin. Medium supplementation during the growth phase increased the cell growth rate from 0.357 x 10(4) to 0.565 x 10(4) cells/mL center dot h. In addition, cholesterol and albumin supplementation increased the expression of ZIKV structural proteins during infection. Higher MOIs led to increased substrate uptake and metabolite production, suggesting intensified cellular metabolism. Western blot analysis revealed that under nonsupplemented conditions, the highest MOI resulted in increased ZIKV envelope production, with a maximum protein concentration range of 1.049 mg/L higher when comparing 6 to 2 MOI via SDS-PAGE densitometry. However, a lower MOI, 2 PFU/cell , might be advantageous when a supplemented medium is used, which upper limit for ZIKV envelope protein concentration was 1.834 higher than that from the nonsupplemented assay in semiquantitative analysis, which reached 23.504 mg/L of ZIKV envelope protein. The resulting VLP had an average diameter of similar to 60 nm, making them suitable for vaccine applications. (AU)

• Processo FAPESP: 22/02713-3 - Estabelecimento de bioprocessos escalonáveis para produção de partículas semelhantes a vírus
• Beneficiário: Eutimio Gustavo Fernández Núñez
• Modalidade de apoio: Auxílio à Pesquisa - Projeto Inicial
• Processo FAPESP: 23/14085-0 - Monitoramento on-line do processo de produção em biorreator de VLP de SARS-CoV-2 por espectroscopia Raman
• Beneficiário: Milena Miyu Teruya
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 23/17480-7 - Otimização da produção de VLP de vírus Zika em meio quimicamente definido utilizando os princípios de Quality by Design
• Beneficiário: Vinícius Aragão Tejo Dias
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 23/09463-5 - Comprimento de onda do laser e condicionamento da amostra no monitoramento bioquímico por espectroscopia Raman da produção de VLP de SARS-CoV-2
• Beneficiário: Júlia Públio Rabello
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica