Sebastian‐Juan Reyes, Phuong Lan Pham, Yves Durocher and Olivier Henry
Article (2024)
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Abstract
Technology scale-up and transfer are a fundamental and critical part of process development in biomanufacturing. Important bioreactor hydrodynamic characteristics such as working volume, overhead gas flow rate, volumetric power input (P/V), impeller type, agitation regimen, sparging aeration strategy, sparger type, and kLa must be selected based on key performance indicators (KPI) to ensure a smooth and seamless process scale-up and transfer. Finding suitable operational setpoints and developing an efficient feeding regimen to ensure process efficacy and consistency are instrumental. In this investigation, process development of a cumate inducible Chinese hamster ovary (CHO) stable pool expressing trimeric SARS-CoV-2 spike protein in 1.8 L benchtop stirred-tank bioreactors is detailed. Various dissolved oxygen levels and aeration air caps were studied to determine their impact on cell growth and metabolism, culture longevity, and endpoint product titers. Once hydrodynamic conditions were tuned to an optimal zone, various feeding strategies were explored to increase culture performance. Dynamic feedings such as feeding based on current culture volume, viable cell density (VCD), oxygen uptake rate (OUR), and bio-capacitance signals were tested and compared to standard bolus addition. Increases in integral of viable cell concentration (IVCC) (1.25-fold) and protein yield (2.52-fold), as well as greater culture longevity (extension of 5 days) were observed in dynamic feeding strategies when compared to periodic bolus feeding. Our study emphasizes the benefits of designing feeding strategies around metabolically relevant signals such as OUR and bio-capacitance signals.
Uncontrolled Keywords
bio-capacitance; CHO stable pool; fed-batch process development; feeding strategy; oxygen uptake rate; SARS-CoV-2 vaccine antigen
Subjects: | 1800 Chemical engineering > 1800 Chemical engineering |
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Department: | Department of Chemical Engineering |
Funders: | NSERC / CRSNG, National Research Council Canada |
Grant number: | RGPIN‐40482021, PR‐023‐1 |
PolyPublie URL: | https://publications.polymtl.ca/59448/ |
Journal Title: | Biotechnology Progress |
Publisher: | Wiley |
DOI: | 10.1002/btpr.3507 |
Official URL: | https://doi.org/10.1002/btpr.3507 |
Date Deposited: | 13 Nov 2024 14:47 |
Last Modified: | 14 Nov 2024 21:32 |
Cite in APA 7: | Reyes, S.‐J., Pham, P. L., Durocher, Y., & Henry, O. (2024). CHO stable pool fed-batch process development of SARS-CoV-2 spike protein production: Impact of aeration conditionsand feeding strategies. Biotechnology Progress, 3507 (20 pages). https://doi.org/10.1002/btpr.3507 |
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