Genome‐Scale Modeling of CHO Cells Unravel the Critical Role of Asparagine in Cell Culture Feed Media

Genome‐Scale Modeling of CHO Cells Unravel the Critical Role of Asparagine in Cell Culture Feed Media

Biotechnology Journal

10.1002/biot.202400072

https://doi.org/10.1002/biot.202400072

Kuin Tian Pang, Yi Fan Hong, Fumi Shozui, Shunpei Furomitsu, Matthew Myint, Ying Swan Ho, Yaron R. Silberberg, Ian Walsh, Meiyappan Lakshmanan

08 November 2024

Pang, K. T., Hong, Y. F., Shozui, F., Furomitsu, S., Myint, M., Ho, Y. S., Silberberg, Y. R., Walsh, I., & Lakshmanan, M. (2024). Genome‐Scale Modeling of CHO Cells Unravel the Critical Role of Asparagine in Cell Culture Feed Media. Biotechnology Journal, 19(11). Portico. https://doi.org/10.1002/biot.202400072

Amino acids, including asparagine, aspartate, glutamine, and glutamate, play important roles in purine and pyrimidine biosynthesis as well as serve as anaplerotic sources fueling the tricarboxylic acid (TCA) cycle for mitochondrial energy generation. Despite extensive studies on glutamine and glutamate in CHO cell cultures, the roles of asparagine and aspartate, especially in feed media, remain underexplored. In this study, we utilized a CHO genome scale model to first deeply characterize the intracellular metabolic states of CHO cells cultured in different combinations of basal and feed media to understand the traits of asparagine/aspartate‐dependent and glutamate‐dependent feeds. Subsequently, we identified the critical role of asparagine and aspartate in the feed media as anaplerotic sources and conducted in silico simulations to ascertain their optimal ratios to improve cell culture performance. Finally, based on the model simulations, we reformulated the feed media by tailoring the concentrations of asparagine and aspartate. Our experimental data reveal a CHO cell preference for asparagine compared with aspartate, and thus maintaining an optimal ratio of these amino acids is a key factor for achieving optimal CHO cell culture performance in biopharmaceutical production.

Publisher Copyright

This research is supported by core funding from: Biomedical Research Council
Grant Reference no. : SC17/21-304412

This research / project is supported by the Agency for Science, Technology, and Research - A*STAR Career Development Fund
Grant Reference no. : C210112057

This is the peer reviewed version of the following article: Pang, K. T., Hong, Y. F., Shozui, F., Furomitsu, S., Myint, M., Ho, Y. S., Silberberg, Y. R., Walsh, I., & Lakshmanan, M. (2024). Genome‐Scale Modeling of CHO Cells Unravel the Critical Role of Asparagine in Cell Culture Feed Media. Biotechnology Journal, 19(11). Portico. https://doi.org/10.1002/biot.202400072, which has been published in final form at https://doi.org/10.1002/biot.202400072. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages thereof by third parties from platforms, services and websites other than Wiley Online Library must be prohibited.

https://oar.a-star.edu.sg/communities-collections/articles/21588

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Bioprocessing Technology Institute