Performance Comparison of Recombinant Baculovirus and Rabies Virus-like Particles production Using Two Culture Platforms

• Published in: Vaccines (Basel) Vol. 11; no. 1; p. 39
• Main Authors: Guardalini, Luis Giovani Oliveira, Cavalcante, Paulo Eduardo da Silva, Leme, Jaci, Mello, Renata Gois de, Bernardino, Thaissa Consoni, Jared, Simone Gonçalves Silva, Antoniazzi, Marta Maria, Astray, Renato Mancini, Tonso, Aldo, Fernández Núñez, Eutimio Gustavo, Jorge, Soraia Attie Calil
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 24.12.2022; MDPI
• Subjects: Assaying; Baculovirus; Bioreactors; Cell culture; Cell death; Cell density; Cell viability; Design of experiments; Disease prevention; Enzyme-linked immunosorbent assay; Equivalence; Flasks; Hepatitis; Infections; Insect cells; Insects; Multiplicity of infection; Optimization; Proteins; quality by design; Rabies; rabies virus-like particles; recombinant baculovirus; RNA polymerase; Sf9 cells; stirred-tank bioreactor; Transmission electron microscopy; Vaccines; viral infection; Virus-like particles; Viruses; United States > US; New Jersey; stirred-tank bioreactor; quality by design; viral infection; rabies virus-like particles; Sf9 cells; recombinant baculovirus
• DOI: 10.3390/vaccines11010039
• ISSN: 2076-393X

This work aimed to assess, following upstream optimization in Schott flasks, the scalability from this culture platform to a stirred-tank bioreactor in order to yield rabies-recombinant baculovirus, bearing genes of G (BVG) and M (BVM) proteins, and to obtain rabies virus-like particles (VLP) from them, using Sf9 insect cells as a host. Equivalent assays in Schott flasks and a bioreactor were performed to compare both systems and a multivariate statistical approach was also carried out to maximize VLP production as a function of BVG and BVM's multiplicity of infection (MOI) and harvest time (HT). Viable cell density, cell viability, virus titer, BVG and BVM quantification by dot-blot, and BVG quantification by Enzyme-Linked Immunosorbent Assay (ELISA) were monitored throughout the assays. Furthermore, transmission electron microscopy was used to characterize rabies VLP. The optimal combination for maximum VLP expression was BVG and BVM MOI of 2.3 pfu/cell and 5.1 pfu/cell, respectively, and 108 h of harvest time. The current study confirmed that the utilization of Schott flasks and a benchtop bioreactor under the conditions applied herein are equivalent regarding the cell death kinetics corresponding to the recombinant baculovirus infection process in Sf9 cells. According to the results, the hydrodynamic and chemical differences in both systems seem to greatly affect the virus and VLP integrity after release.