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|Title:||Proteomic investigation of metabolic shift in mammalian cell culture|
|Authors:||Seow, T.K. |
|Citation:||Seow, T.K., Korke, R., Liang, R.C.M.Y., Ong, S.-E., Ou, K., Wong, K., Hu, W.-S., Chung, M.C.M. (2001). Proteomic investigation of metabolic shift in mammalian cell culture. Biotechnology Progress 17 (6) : 1137-1144. ScholarBank@NUS Repository. https://doi.org/10.1021/bp010101g|
|Abstract:||Mammalian cells, under typical cultivation conditions, produce large quantities of lactate and ammonia that affect cell growth adversely and result in low cell concentration. Controlled nutrient feeding to maintain low concentrations of glucose and glutamine reduces metabolite production drastically, altering the metabolism of the cells. This metabolic shift results in higher cell concentration in continuous cultures and does not affect the specific productivity of the cells. We have taken a proteomics approach to investigate the differential protein expression with metabolic shift. Using two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS), we have found at least eight differentially expressed spots; two proteins were down-regulated, and the others were up-regulated with metabolic shift. These included metabolic enzymes, the brain form of phosphoglycerate mutase, which was down-regulated, and the precursor of the 23 kDa subunit of NADH-ubiquinone oxidoreductase, which was up-regulated. Another enzyme, the L1 isozyme of ubiquitin carboxyl-terminal hydrolase, which is involved in protein turnover and degradation, was also up-regulated in the metabolically altered cells. The remaining down-regulated spot had been identified as two isoforms of cytoplasmic actins, while three of the up-regulated spots were viral GAG polyproteins from various murine viruses. An unidentified protein was also up-regulated in the cells with altered metabolic state. This study shows the potential of using a proteomics approach in deciphering the intracellular changes in cells with physiological changes such as metabolism shift. The new insight into cell metabolism afforded by this analysis will greatly facilitate process optimization of continuous cell cultures.|
|Source Title:||Biotechnology Progress|
|Appears in Collections:||Staff Publications|
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