Accelerating HIV-1 VLP production using stable High Five insect cell pools.
High Five cells
bioreactor
fluorescence-activated cell sorting
metabolism
stable cell pool
virus-like particle
Journal
Biotechnology journal
ISSN: 1860-7314
Titre abrégé: Biotechnol J
Pays: Germany
ID NLM: 101265833
Informations de publication
Date de publication:
Apr 2021
Apr 2021
Historique:
revised:
21
10
2020
received:
18
08
2020
pubmed:
29
11
2020
medline:
28
4
2021
entrez:
28
11
2020
Statut:
ppublish
Résumé
Stable cell pools are receiving a renewed interest as a potential alternative system to clonal cell lines. The shorter development timelines and the capacity to achieve high product yields make them an interesting approach for recombinant protein production. In this study, stable High Five cell pools are assessed for the production of a simple protein, mCherry, and the more complex HIV-1 Gag-eGFP virus-like particles (VLPs). Random integration coupled to fluorescence-activated cell sorting (FACS) in suspension conditions is applied to accelerate the stable cell pool generation process and enrich it with high producer cells. This methodology is successfully transferred to a bioreactor for VLP production, resulting in a 2-fold increase in VLP yields with respect to shake flask cultures. In these conditions, maximum viable cell concentration improves by 1.5-fold, and by-product formation is significantly reduced. Remarkably, a global increase in the uptake of amino acids in the Gag-eGFP stable cell pool is observed when compared with parental High Five cells, reflecting the additional metabolic burden associated with VLP production. These results suggest that stable High Five cell pools are a robust and powerful approach to produce VLPs and other recombinant proteins, and put the basis for future studies aiming to scale up this system.
Identifiants
pubmed: 33247883
doi: 10.1002/biot.202000391
doi:
Substances chimiques
Recombinant Proteins
0
Suspensions
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2000391Informations de copyright
© 2020 Wiley-VCH GmbH.
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