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Perfusion process combining low temperature and valeric acid for enhanced recombinant factor VIII production
Juliana Coronel1, Christoph Heinrich2, Sandra Klausing2
1Federal University of Rio de Janeiro (UFRJ), COPPE, Cell Culture Engineering Laboratory, Rio de Janeiro, RJ, Brazil.
Biotechnology Progress
|October 7, 2019
Summary
This study optimized perfusion cultivation for human factor VIII (FVIII) production by combining mild hypothermia (31°C) with valeric acid (VA). This strategy significantly boosted FVIII production and biological activity in CHO cells, offering a cost-effective method for biopharmaceutical manufacturing.
Area of Science:
- Biopharmaceutical Manufacturing
- Cell Culture Technology
- Bioprocess Engineering
Background:
- Perfusion cultivation is a key method for producing labile and stable biopharmaceuticals.
- Process development in perfusion aims to maximize production capacity for biologics like monoclonal antibodies and blood factors.
- Optimizing conditions for human factor VIII (FVIII) production is crucial for therapeutic applications.
Purpose of the Study:
- To investigate the impact of perfusion cultivation conditions on process productivity for human factor VIII (FVIII) production.
- To evaluate the effects of reduced temperature (31°C) and valeric acid (VA) supplementation on FVIII production in recombinant CHO cells.
- To assess the potential for increasing FVIII specific productivity (qₚ) and biological activity.
Main Methods:
- Recombinant CHO cells were cultivated in bioreactors with inclined settlers under perfusion mode.
- The effects of mild hypothermia (31°C) and valeric acid supplementation were systematically evaluated.
- Cell specific productivity (qₚ) and FVIII biological activity were measured and compared to control conditions (37°C).
Main Results:
- A 2.4-fold increase in FVIII concentration and a 3.0-fold increase in FVIII biological activity were achieved at 31°C with VA supplementation.
- Mild hypothermia combined with VA positively impacted FVIII biological activity, a critical quality attribute.
- Enhanced product transcript levels at low temperatures suggest increased mRNA levels contribute to higher qₚ.
- A high-producer subclone showed 6-fold and 15-fold increases in qₚ under optimized conditions.
Conclusions:
- Combining mild hypothermia (31°C) with valeric acid supplementation is an effective strategy to enhance FVIII production in perfusion culture.
- This perfusion strategy increases product formation without raising production costs.
- The findings are potentially applicable to the perfusion production of other CHO-derived biopharmaceuticals.
- This study presents the first report on the benefits of combining mild hypothermia and VA supplementation in perfusion culture for biopharmaceutical production.
