Multivariate data analysis of process parameters affecting the growth and productivity of stable Chinese hamster ovary cell pools expressing SARS-CoV-2 spike protein as vaccine antigen in early process development.
CHO stable pool
Cumate induction
MVDA
SARS‐CoV‐2 trimeric spike protein
fed‐batch bioreactor production
random forest
Journal
Biotechnology progress
ISSN: 1520-6033
Titre abrégé: Biotechnol Prog
Pays: United States
ID NLM: 8506292
Informations de publication
Date de publication:
25 Apr 2024
25 Apr 2024
Historique:
revised:
25
03
2024
received:
05
01
2024
accepted:
28
03
2024
medline:
25
4
2024
pubmed:
25
4
2024
entrez:
25
4
2024
Statut:
aheadofprint
Résumé
The recent COVID-19 pandemic revealed an urgent need to develop robust cell culture platforms which can react rapidly to respond to this kind of global health issue. Chinese hamster ovary (CHO) stable pools can be a vital alternative to quickly provide gram amounts of recombinant proteins required for early-phase clinical assays. In this study, we analyze early process development data of recombinant trimeric spike protein Cumate-inducible manufacturing platform utilizing CHO stable pool as a preferred production host across three different stirred-tank bioreactor scales (0.75, 1, and 10 L). The impact of cell passage number as an indicator of cell age, methionine sulfoximine (MSX) concentration as a selection pressure, and cell seeding density was investigated using stable pools expressing three variants of concern. Multivariate data analysis with principal component analysis and batch-wise unfolding technique was applied to evaluate the effect of critical process parameters on production variability and a random forest (RF) model was developed to forecast protein production. In order to further improve process understanding, the RF model was analyzed with Shapley value dependency plots so as to determine what ranges of variables were most associated with increased protein production. Increasing longevity, controlling lactate build-up, and altering pH deadband are considered promising approaches to improve overall culture outcomes. The results also demonstrated that these pools are in general stable expressing similar level of spike proteins up to cell passage 11 (~31 cell generations). This enables to expand enough cells required to seed large volume of 200-2000 L bioreactor.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e3467Subventions
Organisme : Natural Sciences and Engineering Research Council of Canada, NSERC-CREATE PrEEmiuM
ID : RGPIN/4048-2021
Organisme : National Research Council Canada
ID : PR-023-1
Informations de copyright
© 2024 National Research Council Canada. Biotechnology Progress published by Wiley‐VCH GmbH on behalf of American Institute of Chemical Engineers. Reproduced with the permission of the Minister of Innovation, Science, and Economic Development.
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