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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

Sebastian‐Juan Reyes, Lucas Lemire, Marjolaine Roy, Hélène LʹÉcuyer-Coelho, Yuliya Martynova, Brian Cass, Robert Voyer, Yves Durocher, Olivier Henry, Phuong Lan Pham and Raul-Santiago Molina

Article (2024)

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Abstract

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.

Uncontrolled Keywords

CHO stable pool; cumate induction; fed-batch bioreactor production; MVDA; random forest; SARS-CoV-2 trimeric spike protein

Subjects: 1800 Chemical engineering > 1800 Chemical engineering
1800 Chemical engineering > 1802 Biochemical engineering
1900 Biomedical engineering > 1900 Biomedical engineering
Department: Department of Chemical Engineering
Funders: NSERC / CRSNG, National Research Council Canada, NSERC / CRSNG - CREATE PrEEmiuM Program
Grant number: RGPIN/4048‐2021, PR‐023‐1
PolyPublie URL: https://publications.polymtl.ca/58209/
Journal Title: Biotechnology Progress
Publisher: Wiley & Sons
DOI: 10.1002/btpr.3467
Official URL: https://doi.org/10.1002/btpr.3467
Date Deposited: 21 May 2024 10:54
Last Modified: 22 May 2024 09:47
Cite in APA 7: Reyes, S.‐J., Lemire, L., Roy, M., LʹÉcuyer-Coelho, H., Martynova, Y., Cass, B., Voyer, R., Durocher, Y., Henry, O., Pham, P. L., & Molina, R.-S. (2024). 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. Biotechnology Progress, e3467 (19 pages). https://doi.org/10.1002/btpr.3467

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