Towards industrial application of quasi real-time metabolic flux analysis for mammalian cell culture

被引:17
作者
Goudar, Chetan
Biener, Richard
Zhang, Chun
Michaels, James
Piret, James
Konstantinov, Konstantin
机构
[1] Bayer HealthCare, Res & Dev, Proc Sci, Biol Prod Div, Albany, CA 94710 USA
[2] Univ British Columbia, Michael Smith Labs, Vancouver, BC V6T 1Z4, Canada
[3] Univ British Columbia, Dept Chem & Biol Engn, Vancouver, BC V6T 1Z4, Canada
[4] Bayer AG, Technol Serv, D-51368 Leverkusen, Germany
[5] Univ Appl Sci Esslingen, Dept Nat Sci, D-73728 Esslingen, Germany
[6] BioMarin Pharmaceut Inc, Novato, CA 94949 USA
来源
CELL CULTURE ENGINEERING | 2006年 / 101卷
关键词
cell culture; metabolic flux; perfusion; quasi real-time analysis; steady-state multiplicity;
D O I
10.1007/10_020
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
Cellular physiology and metabolism were monitored using a quasi real-time combination of on-line and off-line data to estimate metabolic fluxes in an established bioreaction network. The utility of this approach towards optimizing bioreactor operation was demonstrated for CHO cells cultivated in 15 L perfusion reactors at 20 x 10(6) cells/mL. Medium composition and dilution rates were changed to obtain several steady states with varying glucose and glutamine concentrations. When cells were restored to initial culture medium and perfusion rate conditions after being exposed to lower glucose and glutamine concentrations, the pyruvate flux into the TCA cycle was increased 30% while the pyruvate flux through lactate was decreased 30%, suggesting steady-state multiplicity. By appropriately altering cellular metabolism, perfusion bioreactors can operate at lower perfusion rates without significant accumulation of inhibitory metabolites such as lactate. Changes in glucose, lactate and glutamine uptake/production rates had significant effects on the calculation of other fluxes in the network. Sensitivity analysis of these key metabolic fluxes highlighted the need for accurate and reliable real-time sensors. Overall, rapid observation of metabolic fluxes can be a valuable tool for bioprocess development, monitoring and control. The framework presented in this study offers a convenient means for quasi real-time estimation of metabolic fluxes and represents a step towards realizing the potential of metabolic flux analysis for accelerated bioprocess optimization.
引用
收藏
页码:99 / 118
页数:20
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