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Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory Evolution

The use of non-standard culture conditions has proven efficient to increase cell performance and recombinant protein production in different cell hosts. However, the establishment of high-producing cell populations through adaptive laboratory evolution (ALE) has been poorly explored, in particular f...

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Autores principales: Correia, Ricardo, Fernandes, Bárbara, Alves, Paula M., Carrondo, Manuel J.T., Roldão, António
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7711658/
https://www.ncbi.nlm.nih.gov/pubmed/33036359
http://dx.doi.org/10.3390/vaccines8040589
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author Correia, Ricardo
Fernandes, Bárbara
Alves, Paula M.
Carrondo, Manuel J.T.
Roldão, António
author_facet Correia, Ricardo
Fernandes, Bárbara
Alves, Paula M.
Carrondo, Manuel J.T.
Roldão, António
author_sort Correia, Ricardo
collection PubMed
description The use of non-standard culture conditions has proven efficient to increase cell performance and recombinant protein production in different cell hosts. However, the establishment of high-producing cell populations through adaptive laboratory evolution (ALE) has been poorly explored, in particular for insect cells. In this study, insect High Five cells were successfully adapted to grow at a neutral culture pH (7.0) through ALE for an improved production of influenza hemagglutinin (HA)-displaying virus-like particles (VLPs). A stepwise approach was used for the adaptation process, in which the culture pH gradually increased from standard 6.2 to 7.0 (ΔPh = 0.2–0.3), and cells were maintained at each pH value for 2–3 weeks until a constant growth rate and a cell viability over 95% were observed. These adapted cells enabled an increase in cell-specific HA productivity up to three-fold and volumetric HA titer of up to four-fold as compared to non-adapted cells. Of note, the adaptation process is the element driving increased specific HA productivity as a pH shift alone was inefficient at improving productivities. The production of HA-VLPs in adapted cells was successfully demonstrated at the bioreactor scale. The produced HA-VLPs show the typical size and morphology of influenza VLPs, thus confirming the null impact of the adaptation process and neutral culture pH on the quality of HA-VLPs produced. This work strengthens the potential of ALE as a bioprocess engineering strategy to improve the production of influenza HA-VLPs in insect High Five cells.
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spelling pubmed-77116582020-12-04 Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory Evolution Correia, Ricardo Fernandes, Bárbara Alves, Paula M. Carrondo, Manuel J.T. Roldão, António Vaccines (Basel) Article The use of non-standard culture conditions has proven efficient to increase cell performance and recombinant protein production in different cell hosts. However, the establishment of high-producing cell populations through adaptive laboratory evolution (ALE) has been poorly explored, in particular for insect cells. In this study, insect High Five cells were successfully adapted to grow at a neutral culture pH (7.0) through ALE for an improved production of influenza hemagglutinin (HA)-displaying virus-like particles (VLPs). A stepwise approach was used for the adaptation process, in which the culture pH gradually increased from standard 6.2 to 7.0 (ΔPh = 0.2–0.3), and cells were maintained at each pH value for 2–3 weeks until a constant growth rate and a cell viability over 95% were observed. These adapted cells enabled an increase in cell-specific HA productivity up to three-fold and volumetric HA titer of up to four-fold as compared to non-adapted cells. Of note, the adaptation process is the element driving increased specific HA productivity as a pH shift alone was inefficient at improving productivities. The production of HA-VLPs in adapted cells was successfully demonstrated at the bioreactor scale. The produced HA-VLPs show the typical size and morphology of influenza VLPs, thus confirming the null impact of the adaptation process and neutral culture pH on the quality of HA-VLPs produced. This work strengthens the potential of ALE as a bioprocess engineering strategy to improve the production of influenza HA-VLPs in insect High Five cells. MDPI 2020-10-07 /pmc/articles/PMC7711658/ /pubmed/33036359 http://dx.doi.org/10.3390/vaccines8040589 Text en © 2020 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Correia, Ricardo
Fernandes, Bárbara
Alves, Paula M.
Carrondo, Manuel J.T.
Roldão, António
Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory Evolution
title Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory Evolution
title_full Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory Evolution
title_fullStr Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory Evolution
title_full_unstemmed Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory Evolution
title_short Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory Evolution
title_sort improving influenza ha-vlps production in insect high five cells via adaptive laboratory evolution
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7711658/
https://www.ncbi.nlm.nih.gov/pubmed/33036359
http://dx.doi.org/10.3390/vaccines8040589
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