MONITORING HYBRIDOMA METABOLISM IN CONTINUOUS SUSPENSION-CULTURE AT THE INTRACELLULAR LEVEL .1. STEADY-STATE RESPONSES TO DIFFERENT GLUTAMINE FEED CONCENTRATIONS

被引:27
作者
SCHMID, G
KELLER, T
机构
[1] Department PRTM, F. Hoffmann-La Roche Ltd., Pharmaceutical Research-New Technologies, Basel
关键词
HYBRIDOMA METABOLISM; ANTIBODY PRODUCTION; CONTINUOUS CULTURE; INTRACELLULAR METABOLITE CONCENTRATIONS; KREBS CYCLE INTERMEDIATES; AMINO ACID POOLS;
D O I
10.1007/BF02521749
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
A model mouse hybridoma cell line was grown in continuous culture experiments in a serum-free low-protein lipid-free medium. The steady-state responses of cell numbers, extra- and intracellular metabolite concentrations, substrate and (by) product consumption/production rates, and yield coefficients were investigated as a function of step changes in the glutamine concentration of the feed medium. In addition to the commonly performed analysis of metabolites in culture supernatants, we prepared perchloric acid extracts of cells and determined the amount and the composition of intracellular amino acids and organic acids. Significant differences were found with respect to intracellular metabolite pools for cells growing at nearly identical specific growth rates. To our knowledge this is the first time that data on the intracellular concentrations (pools) of amino acids and Krebs cycle intermediates are reported in the literature that were obtained under carefully defined culture conditions such as those attained in continuous culture experiments.
引用
收藏
页码:217 / 229
页数:13
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