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Performance Comparison of Recombinant Baculovirus and Rabies Virus-like Particles production Using Two Culture Platforms
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 t...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9867115/ https://www.ncbi.nlm.nih.gov/pubmed/36679884 http://dx.doi.org/10.3390/vaccines11010039 |
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author | Guardalini, Luis Giovani Oliveira Cavalcante, Paulo Eduardo da Silva Leme, Jaci de Mello, Renata Gois Bernardino, Thaissa Consoni Jared, Simone Gonçalves Silva Antoniazzi, Marta Maria Astray, Renato Mancini Tonso, Aldo Fernández Núñez, Eutimio Gustavo Jorge, Soraia Attie Calil |
author_facet | Guardalini, Luis Giovani Oliveira Cavalcante, Paulo Eduardo da Silva Leme, Jaci de Mello, Renata Gois Bernardino, Thaissa Consoni Jared, Simone Gonçalves Silva Antoniazzi, Marta Maria Astray, Renato Mancini Tonso, Aldo Fernández Núñez, Eutimio Gustavo Jorge, Soraia Attie Calil |
author_sort | Guardalini, Luis Giovani Oliveira |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9867115 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-98671152023-01-22 Performance Comparison of Recombinant Baculovirus and Rabies Virus-like Particles production Using Two Culture Platforms Guardalini, Luis Giovani Oliveira Cavalcante, Paulo Eduardo da Silva Leme, Jaci de Mello, Renata Gois Bernardino, Thaissa Consoni Jared, Simone Gonçalves Silva Antoniazzi, Marta Maria Astray, Renato Mancini Tonso, Aldo Fernández Núñez, Eutimio Gustavo Jorge, Soraia Attie Calil Vaccines (Basel) Article 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. MDPI 2022-12-24 /pmc/articles/PMC9867115/ /pubmed/36679884 http://dx.doi.org/10.3390/vaccines11010039 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Guardalini, Luis Giovani Oliveira Cavalcante, Paulo Eduardo da Silva Leme, Jaci de Mello, Renata Gois Bernardino, Thaissa Consoni Jared, Simone Gonçalves Silva Antoniazzi, Marta Maria Astray, Renato Mancini Tonso, Aldo Fernández Núñez, Eutimio Gustavo Jorge, Soraia Attie Calil Performance Comparison of Recombinant Baculovirus and Rabies Virus-like Particles production Using Two Culture Platforms |
title | Performance Comparison of Recombinant Baculovirus and Rabies Virus-like Particles production Using Two Culture Platforms |
title_full | Performance Comparison of Recombinant Baculovirus and Rabies Virus-like Particles production Using Two Culture Platforms |
title_fullStr | Performance Comparison of Recombinant Baculovirus and Rabies Virus-like Particles production Using Two Culture Platforms |
title_full_unstemmed | Performance Comparison of Recombinant Baculovirus and Rabies Virus-like Particles production Using Two Culture Platforms |
title_short | Performance Comparison of Recombinant Baculovirus and Rabies Virus-like Particles production Using Two Culture Platforms |
title_sort | performance comparison of recombinant baculovirus and rabies virus-like particles production using two culture platforms |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9867115/ https://www.ncbi.nlm.nih.gov/pubmed/36679884 http://dx.doi.org/10.3390/vaccines11010039 |
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