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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| Format: | Article |
| Language: | English |
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MDPI AG
2020-10-01
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| Series: | Vaccines |
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| Online Access: | https://www.mdpi.com/2076-393X/8/4/589 |
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| author | Ricardo Correia Bárbara Fernandes Paula M. Alves Manuel J.T. Carrondo António Roldão |
| author_facet | Ricardo Correia Bárbara Fernandes Paula M. Alves Manuel J.T. Carrondo António Roldão |
| author_sort | Ricardo Correia |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-10T15:48:37Z |
| format | Article |
| id | doaj.art-3f54b88796624ab78b2562867c959eb2 |
| institution | Directory Open Access Journal |
| issn | 2076-393X |
| language | English |
| last_indexed | 2024-03-10T15:48:37Z |
| publishDate | 2020-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj.art-3f54b88796624ab78b2562867c959eb22023-11-20T16:15:59ZengMDPI AGVaccines2076-393X2020-10-018458910.3390/vaccines8040589Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory EvolutionRicardo Correia0Bárbara Fernandes1Paula M. Alves2Manuel J.T. Carrondo3António Roldão4IBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, PortugalIBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, PortugalIBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, PortugalIBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, PortugalIBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, PortugalThe 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.https://www.mdpi.com/2076-393X/8/4/589adaptive laboratory evolutioninsect High Five cellsbaculovirus expression systeminfluenza HA-VLPsimproved production |
| spellingShingle | Ricardo Correia Bárbara Fernandes Paula M. Alves Manuel J.T. Carrondo António Roldão Improving Influenza HA-Vlps Production in Insect High Five Cells via Adaptive Laboratory Evolution Vaccines adaptive laboratory evolution insect High Five cells baculovirus expression system influenza HA-VLPs improved production |
| 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 | adaptive laboratory evolution insect High Five cells baculovirus expression system influenza HA-VLPs improved production |
| url | https://www.mdpi.com/2076-393X/8/4/589 |
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