Amino acid consumption in naive and recombinant CHO cell cultures: producers of a monoclonal antibody

被引:45
作者
Carrillo-Cocom, L. M. [1 ]
Genel-Rey, T. [1 ]
Araiz-Hernandez, D. [1 ]
Lopez-Pacheco, F. [1 ]
Lopez-Meza, J. [1 ]
Rocha-Pizana, M. R. [1 ]
Ramirez-Medrano, A. [1 ]
Alvarez, M. M. [1 ]
机构
[1] FEMSA, Tecnol Monterrey Monterrey, Ctr Biotecnol, Monterrey 64849, Mexico, Mexico
关键词
CHO cells; Amino acid consumption; Monoclonal antibodies; Biopharmaceutical; Feed strategies; Mammalian cell culture; SERUM-FREE MEDIUM; FED-BATCH PROCESS; STATISTICAL DESIGN; HYBRIDOMA CELLS; MEDIA; GLUCOSE; METABOLISM; LACTATE; OPTIMIZATION; IMPROVEMENT;
D O I
10.1007/s10616-014-9720-5
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Most commercial media for mammalian cell culture are designed to satisfy the amino acid requirements for cell growth, but not necessarily those for recombinant protein production. In this study, we analyze the amino acid consumption pattern in na < ve and recombinant Chinese hamster ovary (CHO) cell cultures. The recombinant model we chose was a CHO-S cell line engineered to produce a monoclonal antibody. We report the cell concentration, product concentration, and amino acid concentration profiles in na < ve and recombinant cell cultures growing in CD OptiCHO (TM) medium with or without amino acid supplementation with a commercial supplement (CHO CD EfficientFeed (TM) B). We quantify and discuss the amino acid demands due to cell growth and recombinant protein production during long term fed batch cultivation protocols. We confirmed that a group of five amino acids, constituting the highest mass fraction of the product, shows the highest depletion rates and could become limiting for product expression. In our experiments, alanine, a non-important mass constituent of the product, is in high demand during recombinant protein production. Evaluation of specific amino acid demands could be of great help in the design of feeding/supplementation strategies for recombinant mammalian cell cultures.
引用
收藏
页码:809 / 820
页数:12
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