Cell specific perfusion rates drive growth dynamics and metabolism in CHO N-1 perfusion processes independent of perfusion rate control method

The production of monoclonal antibodies (mAbs) using Chinese Hamster Ovary (CHO) cell host systems often faces challenges in terms of manufacturing costs and efficiency. To address these challenges, process intensification with high seeding density production processes applying N-1 perfusion is util...

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Main Authors: Julia Walther, Tiago Ribeiro da Costa, Lydia Winkler, Jochen Schaub, Tobias Habicher
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Bioengineering and Biotechnology
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Online Access:https://www.frontiersin.org/articles/10.3389/fbioe.2025.1608889/full
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author Julia Walther
Tiago Ribeiro da Costa
Lydia Winkler
Jochen Schaub
Tobias Habicher
author_facet Julia Walther
Tiago Ribeiro da Costa
Lydia Winkler
Jochen Schaub
Tobias Habicher
author_sort Julia Walther
collection DOAJ
description The production of monoclonal antibodies (mAbs) using Chinese Hamster Ovary (CHO) cell host systems often faces challenges in terms of manufacturing costs and efficiency. To address these challenges, process intensification with high seeding density production processes applying N-1 perfusion is utilized. This study delves into the impact of cell specific perfusion rate (CSPR) and the total exchanged medium in relation to the reactor working volume (iVVD) on cell growth dynamics and metabolic stability in N-1 perfusion cultivations. The effect of varying the perfusion rate increase (PRI) while keeping the amount of exchanged medium constant is investigated, revealing a positive correlation between iVVD and overall cell growth. However, this effect plateaus at higher values, indicating diminishing returns on cell growth with increased medium use. We found that CSPR directly influences the specific metabolic rates of several metabolites and amino acids, accelerating overall metabolism without necessarily affecting growth. Interestingly, the specific metabolic rates are driven by the CSPR after a metabolic adaptation until day 2.5. Besides adjusting perfusion rates every 24 h, the potential benefits of real-time CSPR control using a capacitance probe are explored. While real-time control offers more precise regulation of the perfusion rate, growth and metabolic behavior is comparable to predefined rates within the tested range. This study demonstrates that optimization of factors such as CSPR, iVVD, and PRI can lead to improved cell growth and viability with the potential to decrease media expenditure, thereby reducing manufacturing costs for the production of mAbs using CHO cell host systems.
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spelling doaj-art-bc197f9b6e6f4ea4a129c4857088a9102025-08-20T03:28:09ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852025-07-011310.3389/fbioe.2025.16088891608889Cell specific perfusion rates drive growth dynamics and metabolism in CHO N-1 perfusion processes independent of perfusion rate control methodJulia WaltherTiago Ribeiro da CostaLydia WinklerJochen SchaubTobias HabicherThe production of monoclonal antibodies (mAbs) using Chinese Hamster Ovary (CHO) cell host systems often faces challenges in terms of manufacturing costs and efficiency. To address these challenges, process intensification with high seeding density production processes applying N-1 perfusion is utilized. This study delves into the impact of cell specific perfusion rate (CSPR) and the total exchanged medium in relation to the reactor working volume (iVVD) on cell growth dynamics and metabolic stability in N-1 perfusion cultivations. The effect of varying the perfusion rate increase (PRI) while keeping the amount of exchanged medium constant is investigated, revealing a positive correlation between iVVD and overall cell growth. However, this effect plateaus at higher values, indicating diminishing returns on cell growth with increased medium use. We found that CSPR directly influences the specific metabolic rates of several metabolites and amino acids, accelerating overall metabolism without necessarily affecting growth. Interestingly, the specific metabolic rates are driven by the CSPR after a metabolic adaptation until day 2.5. Besides adjusting perfusion rates every 24 h, the potential benefits of real-time CSPR control using a capacitance probe are explored. While real-time control offers more precise regulation of the perfusion rate, growth and metabolic behavior is comparable to predefined rates within the tested range. This study demonstrates that optimization of factors such as CSPR, iVVD, and PRI can lead to improved cell growth and viability with the potential to decrease media expenditure, thereby reducing manufacturing costs for the production of mAbs using CHO cell host systems.https://www.frontiersin.org/articles/10.3389/fbioe.2025.1608889/fullcell specific perfusion rate (CSPR)N-1 perfusion cultivationsspecific metabolic ratescell growth dynamicsmedium exchangeCHO cell cultivation
spellingShingle Julia Walther
Tiago Ribeiro da Costa
Lydia Winkler
Jochen Schaub
Tobias Habicher
Cell specific perfusion rates drive growth dynamics and metabolism in CHO N-1 perfusion processes independent of perfusion rate control method
Frontiers in Bioengineering and Biotechnology
cell specific perfusion rate (CSPR)
N-1 perfusion cultivations
specific metabolic rates
cell growth dynamics
medium exchange
CHO cell cultivation
title Cell specific perfusion rates drive growth dynamics and metabolism in CHO N-1 perfusion processes independent of perfusion rate control method
title_full Cell specific perfusion rates drive growth dynamics and metabolism in CHO N-1 perfusion processes independent of perfusion rate control method
title_fullStr Cell specific perfusion rates drive growth dynamics and metabolism in CHO N-1 perfusion processes independent of perfusion rate control method
title_full_unstemmed Cell specific perfusion rates drive growth dynamics and metabolism in CHO N-1 perfusion processes independent of perfusion rate control method
title_short Cell specific perfusion rates drive growth dynamics and metabolism in CHO N-1 perfusion processes independent of perfusion rate control method
title_sort cell specific perfusion rates drive growth dynamics and metabolism in cho n 1 perfusion processes independent of perfusion rate control method
topic cell specific perfusion rate (CSPR)
N-1 perfusion cultivations
specific metabolic rates
cell growth dynamics
medium exchange
CHO cell cultivation
url https://www.frontiersin.org/articles/10.3389/fbioe.2025.1608889/full
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