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Article

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

1
IBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal
2
ITQB NOVA, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal
*
Author to whom correspondence should be addressed.
Vaccines 2020, 8(4), 589; https://doi.org/10.3390/vaccines8040589
Received: 7 September 2020 / Revised: 28 September 2020 / Accepted: 5 October 2020 / Published: 7 October 2020
(This article belongs to the Section Influenza Virus Vaccines)
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. View Full-Text
Keywords: adaptive laboratory evolution; insect High Five cells; baculovirus expression system; influenza HA-VLPs; improved production adaptive laboratory evolution; insect High Five cells; baculovirus expression system; influenza HA-VLPs; improved production
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MDPI and ACS Style

Correia, R.; Fernandes, B.; Alves, P.M.; Carrondo, M.J.T.; Roldão, A. Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory Evolution. Vaccines 2020, 8, 589. https://doi.org/10.3390/vaccines8040589

AMA Style

Correia R, Fernandes B, Alves PM, Carrondo MJT, Roldão A. Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory Evolution. Vaccines. 2020; 8(4):589. https://doi.org/10.3390/vaccines8040589

Chicago/Turabian Style

Correia, Ricardo, Bárbara Fernandes, Paula M. Alves, Manuel J.T. Carrondo, and António Roldão. 2020. "Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory Evolution" Vaccines 8, no. 4: 589. https://doi.org/10.3390/vaccines8040589

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