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Sf9 Cells Metabolism and Viability When Coinfected with Two Monocistronic Baculoviruses to Produce Rabies Virus-like Particles

ABSTRACT: This work aimed to describe the dynamics of the Sf9 insect cells death and primary metabolism when this host is infected simultaneously by two recombinant baculoviruses (BV) expressing rabies glycoprotein (BVG) and matrix protein (BVM) genes to produce rabies virus-like particles (VLP) at...

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Autores principales: Leme, Jaci, Guardalini, Luis Giovani Oliveira, Bernardino, Thaissa Consoni, Astray, Renato Mancini, Tonso, Aldo, Núñez, Eutimio Gustavo Fernández, Jorge, Soraia Attie Calil
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer US 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9672645/
https://www.ncbi.nlm.nih.gov/pubmed/36396754
http://dx.doi.org/10.1007/s12033-022-00586-x
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author Leme, Jaci
Guardalini, Luis Giovani Oliveira
Bernardino, Thaissa Consoni
Astray, Renato Mancini
Tonso, Aldo
Núñez, Eutimio Gustavo Fernández
Jorge, Soraia Attie Calil
author_facet Leme, Jaci
Guardalini, Luis Giovani Oliveira
Bernardino, Thaissa Consoni
Astray, Renato Mancini
Tonso, Aldo
Núñez, Eutimio Gustavo Fernández
Jorge, Soraia Attie Calil
author_sort Leme, Jaci
collection PubMed
description ABSTRACT: This work aimed to describe the dynamics of the Sf9 insect cells death and primary metabolism when this host is infected simultaneously by two recombinant baculoviruses (BV) expressing rabies glycoprotein (BVG) and matrix protein (BVM) genes to produce rabies virus-like particles (VLP) at different multiplicities of infection (MOI). Schott flasks essays covering a wide range of MOI for both BV were performed. Viable cell density, cell viability, glucose, glutamine, glutamate, lactate, ammonium, and rabies proteins concentrations were monitored over the infection phase. The expression of both recombinant proteins was not limited by glucose, glutamine, and glutamate in a broad MOI (pfu/cell) range of BVG (0.15–12.5) and BVM (0.1–5.0) using SF900 III serum free culture medium. Death phase initiation and the specific death rate depend on BV MOI. The wave pattern of nutrient/metabolite profiles throughout the viral infection phase is related to the baculovirus lytic cycle. The optimal MOIs ratio between BVG (2.5–4.5) and BVM (1.0–3.0) for maximum protein expression was defined. The produced rabies VLP sizes are close to 78 nm. In general, these work outputs bring a better understanding of the metabolic performance of Sf9 cells when infected by BV for producing VLP, and specifically, for progressing in a rabies VLP vaccine development. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12033-022-00586-x.
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spelling pubmed-96726452022-11-18 Sf9 Cells Metabolism and Viability When Coinfected with Two Monocistronic Baculoviruses to Produce Rabies Virus-like Particles Leme, Jaci Guardalini, Luis Giovani Oliveira Bernardino, Thaissa Consoni Astray, Renato Mancini Tonso, Aldo Núñez, Eutimio Gustavo Fernández Jorge, Soraia Attie Calil Mol Biotechnol Original Paper ABSTRACT: This work aimed to describe the dynamics of the Sf9 insect cells death and primary metabolism when this host is infected simultaneously by two recombinant baculoviruses (BV) expressing rabies glycoprotein (BVG) and matrix protein (BVM) genes to produce rabies virus-like particles (VLP) at different multiplicities of infection (MOI). Schott flasks essays covering a wide range of MOI for both BV were performed. Viable cell density, cell viability, glucose, glutamine, glutamate, lactate, ammonium, and rabies proteins concentrations were monitored over the infection phase. The expression of both recombinant proteins was not limited by glucose, glutamine, and glutamate in a broad MOI (pfu/cell) range of BVG (0.15–12.5) and BVM (0.1–5.0) using SF900 III serum free culture medium. Death phase initiation and the specific death rate depend on BV MOI. The wave pattern of nutrient/metabolite profiles throughout the viral infection phase is related to the baculovirus lytic cycle. The optimal MOIs ratio between BVG (2.5–4.5) and BVM (1.0–3.0) for maximum protein expression was defined. The produced rabies VLP sizes are close to 78 nm. In general, these work outputs bring a better understanding of the metabolic performance of Sf9 cells when infected by BV for producing VLP, and specifically, for progressing in a rabies VLP vaccine development. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12033-022-00586-x. Springer US 2022-11-18 2023 /pmc/articles/PMC9672645/ /pubmed/36396754 http://dx.doi.org/10.1007/s12033-022-00586-x Text en © The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2022, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Original Paper
Leme, Jaci
Guardalini, Luis Giovani Oliveira
Bernardino, Thaissa Consoni
Astray, Renato Mancini
Tonso, Aldo
Núñez, Eutimio Gustavo Fernández
Jorge, Soraia Attie Calil
Sf9 Cells Metabolism and Viability When Coinfected with Two Monocistronic Baculoviruses to Produce Rabies Virus-like Particles
title Sf9 Cells Metabolism and Viability When Coinfected with Two Monocistronic Baculoviruses to Produce Rabies Virus-like Particles
title_full Sf9 Cells Metabolism and Viability When Coinfected with Two Monocistronic Baculoviruses to Produce Rabies Virus-like Particles
title_fullStr Sf9 Cells Metabolism and Viability When Coinfected with Two Monocistronic Baculoviruses to Produce Rabies Virus-like Particles
title_full_unstemmed Sf9 Cells Metabolism and Viability When Coinfected with Two Monocistronic Baculoviruses to Produce Rabies Virus-like Particles
title_short Sf9 Cells Metabolism and Viability When Coinfected with Two Monocistronic Baculoviruses to Produce Rabies Virus-like Particles
title_sort sf9 cells metabolism and viability when coinfected with two monocistronic baculoviruses to produce rabies virus-like particles
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9672645/
https://www.ncbi.nlm.nih.gov/pubmed/36396754
http://dx.doi.org/10.1007/s12033-022-00586-x
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