Debottlenecking and reformulating feed media for improved CHO cell growth and titer by data-driven and model-guided analyses.

Autor: Park SY; School of Chemical Engineering, Sungkyunkwan University, Suwon, Gyeonggi-do, Republic of Korea., Choi DH; School of Chemical Engineering, Sungkyunkwan University, Suwon, Gyeonggi-do, Republic of Korea., Song J; School of Chemical Engineering, Sungkyunkwan University, Suwon, Gyeonggi-do, Republic of Korea., Park U; Ajinomoto Genexine Co., Ltd., CELLiST Solution Center, Incheon, Republic of Korea., Cho H; Ajinomoto Genexine Co., Ltd., CELLiST Solution Center, Incheon, Republic of Korea., Hong BH; Ajinomoto Genexine Co., Ltd., CELLiST Solution Center, Incheon, Republic of Korea., Silberberg YR; Ajinomoto Genexine Co., Ltd., CELLiST Solution Center, Incheon, Republic of Korea., Lee DY; School of Chemical Engineering, Sungkyunkwan University, Suwon, Gyeonggi-do, Republic of Korea.; Bitwinners Pte. Ltd., Singapore.
Jazyk: angličtina
Zdroj: Biotechnology journal [Biotechnol J] 2023 Dec; Vol. 18 (12), pp. e2300126. Date of Electronic Publication: 2023 Aug 27.
DOI: 10.1002/biot.202300126
Abstrakt: Designing and selecting cell culture media along with their feeding are a key strategy to maximize culture performance in biopharmaceutical processes. However, the sensitivity of mammalian cells to their culture environment necessitates specific nutritional requirements for their growth and the production of high-quality proteins such as antibodies, depending on the cell lines and operational conditions employed. In this regard, previously we developed a data-driven and in-silico model-guided systematic framework to investigate the effect of growth media on Chinese hamster ovary (CHO) cell culture performance, allowing us to design and reformulate basal media. To expand our exploration for media development research, we evaluated two chemically defined feed media, A and B, using a monoclonal antibody-producing CHO-K1 cell line in ambr15 bioreactor runs. We observed a significant impact of the feed media on various aspects of cell culture, including growth, longevity, viability, productivity, and the production of toxic metabolites. Specifically, the concentrated feed A was inadequate in sustaining prolonged cell culture and achieving high titers when compared to feed B. Within our framework, we systematically investigated the major metabolic bottlenecks in the tricarboxylic acid cycle and relevant amino acid transferase reactions. This analysis identified target components that play a crucial role in alleviating bottlenecks and designing highly productive cell cultures, specifically the addition of glutamate to feed A and asparagine to feed B. Based on our findings, we reformulated the feeds by adjusting the amounts of the targeted amino acids and successfully validated the effectiveness of the strategy in promoting cell growth, life span, and/or titer.
(© 2023 Wiley-VCH GmbH.)
Databáze: MEDLINE
Externí odkaz: