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Zika virus-like particle production in a stirred tank bioreactor using a baculovirus/insect cell system

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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
Total Authors: 11
Document type: Journal article
Source: Bioprocess and Biosystems Engineering; v. 48, n. 12, p. 16-pg., 2025-09-22.
Abstract

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)

FAPESP's process: 22/02713-3 - Establishment of scalable bioprocesses for producing virus-like particles
Grantee:Eutimio Gustavo Fernández Núñez
Support Opportunities: Research Grants - Initial Project
FAPESP's process: 23/14085-0 - Online monitoring of the SARS-CoV-2 VLP bioreactor production process by Raman spectroscopy
Grantee:Milena Miyu Teruya
Support Opportunities: Scholarships in Brazil - Scientific Initiation
FAPESP's process: 23/17480-7 - Optimization of Zika virus VLP production in chemically defined medium using Quality by Design principles
Grantee:Vinícius Aragão Tejo Dias
Support Opportunities: Scholarships in Brazil - Master
FAPESP's process: 23/09463-5 - Laser wavelength and sample conditioning effects on biochemical monitoring of SARS-CoV-2 VLP production upstream stage by Raman spectroscopy
Grantee:Júlia Públio Rabello
Support Opportunities: Scholarships in Brazil - Scientific Initiation