Metabolomic characterization of monoclonal antibody-producing Chinese hamster lung (CHL)-YN cells in glucose-controlled serum-free fed-batch operation.

Autor: Sukwattananipaat P; Graduate School of Engineering, Osaka University, Osaka, Japan., Kuroda H; Graduate School of Engineering, Osaka University, Osaka, Japan.; Shimadzu Corp., Kyoto, Japan.; Shimadzu Analytical Innovation Research Laboratories, Osaka, Japan., Yamano-Adachi N; Graduate School of Engineering, Osaka University, Osaka, Japan.; Institute for Open and Transdisciplinary Research Initiatives, Osaka University, Osaka, Japan.; Manufacturing Technology Association of Biologics (MAB), Hyogo, Japan., Omasa T; Graduate School of Engineering, Osaka University, Osaka, Japan.; Institute for Open and Transdisciplinary Research Initiatives, Osaka University, Osaka, Japan.; Manufacturing Technology Association of Biologics (MAB), Hyogo, Japan.
Jazyk: angličtina
Zdroj: Biotechnology and bioengineering [Biotechnol Bioeng] 2024 Sep; Vol. 121 (9), pp. 2848-2867. Date of Electronic Publication: 2024 Jun 22.
DOI: 10.1002/bit.28777
Abstrakt: The fast-growing Chinese hamster lung (CHL)-YN cell line was recently developed for monoclonal antibody production. In this study, we applied a serum-free fed-batch cultivation process to immunoglobulin (Ig)G1-producing CHL-YN cells, which were then used to design a dynamic glucose supply system to stabilize the extracellular glucose concentration based on glucose consumption. Glucose consumption of the cultures rapidly oscillated following three phases of glutamine metabolism: consumption, production, and re-consumption. Use of the dynamic glucose supply prolonged the viability of the CHL-YN-IgG1 cell cultures and increased IgG1 production. Liquid chromatography with tandem mass spectrometry-based target metabolomics analysis of the extracellular metabolites during the first glutamine shift was conducted to search for depleted compounds. The results suggest that the levels of four amino acids, namely arginine, aspartate, methionine, and serine, were sharply decreased in CHL-YN cells during glutamine production. Supporting evidence from metabolic and gene expression analyses also suggest that CHL-YN cells acquired ornithine- and cystathionine-production abilities that differed from those in Chinese hamster ovary-K1 cells, potentially leading to proline and cysteine biosynthesis.
(© 2024 The Author(s). Biotechnology and Bioengineering published by Wiley Periodicals LLC.)
Databáze: MEDLINE
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