Abstract
To better understand intracellular responses rCHO cells expressing an antibody to hyperosmotic pressure, we have taken a proteomics approach. Using two-dimensional electrophoresis and mass spectrometry, a proteome profile of rCHO cells comprising 23 identified proteins was established. Based on this proteome profile, we found 3 proteins of which expression levels were significantly changed at 450 mOsm/kg. Compared to the results at 300 mOsm/kg, two glycolytic enzymes, glyceraldehyde-3-phosphate dehydrogenase and pyruvate kinase, were found to be up-regulated, probably leading to an increased metabolic energy for antibody synthesis. The elevation of specific glucose consumption rate at 450 mOsm/kg agreed with the up-regulation of these glycolytic enzymes. On the other hand, tubulin expression was down-regulated, reflecting a depressed cell growth rate at 450 mOsm/kg. Taken together, this study shows the potential of the proteomics approach in understanding intracellular and physiological changes in cells and seeking a better insight into possible environmental or genetic manipulation approaches for increasing foreign protein production in rCHO cells.
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4. References
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Lee, M.S., Kim, K.W., Kim, Y.H., Lee, G.M. (2005). Proteome Analysis of Recombinant CHO Cells Under Hyperosmotic Stress. In: Gòdia, F., Fussenegger, M. (eds) Animal Cell Technology Meets Genomics. ESACT Proceedings, vol 2. Springer, Dordrecht. https://doi.org/10.1007/1-4020-3103-3_10
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DOI: https://doi.org/10.1007/1-4020-3103-3_10
Publisher Name: Springer, Dordrecht
Print ISBN: 978-1-4020-2791-8
Online ISBN: 978-1-4020-3103-8
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