1. Isotope labeling to determine the dynamics of metabolic response in CHO cell perfusion bioreactors using MALDI-TOF-MS.
- Author
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Karst, Daniel J., Steinhoff, Robert F., Kopp, Marie R. G., Soos, Miroslav, Zenobi, Renato, and Morbidelli, Massimo
- Subjects
RADIOLABELING ,BIOREACTORS ,CHO cell ,CELL metabolism ,HYDRODYNAMICS - Abstract
The steady-state operation of Chinese hamster ovary (CHO) cells in perfusion bioreactors requires the equilibration of reactor dynamics and cell metabolism. Accordingly, in this work we investigate the transient cellular response to changes in its environment and their interactions with the bioreactor hydrodynamics. This is done in a benchtop perfusion bioreactor using MALDI-TOF MS through isotope labeling of complex intracellular nucleotides (ATP, UTP) and nucleotide sugars (UDP-Hex, UDP-HexNAc). By switching to a
13 C6 glucose containing feed media during constant operation at 20 × 106 cells and a perfusion rate of 1 reactor volume per day, isotopic steady state was studied. A step change to the13 C6 glucose medium in spin tubes allowed the determination of characteristic times for the intracellular turnover of unlabeled metabolites pools [ABSTRACT FROM AUTHOR]- Published
- 2017
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