Variation in the Ratios and Concentrations of Nucleotide Triphosphates and Udp-Sugars During a Perfused Batch Cultivation of Hybridoma Cells
A murine hybridoma cell line, which produces a monoclonal murine IgG2a antibody, was cultivated in a stirred reactor equipped with bubble-free aeration and a continuous perfusion system. During growth of up to 3.1•107 viable cells per ml, intracellular amounts of ATP, ADP, AMP, NAD, GTP, UTP, UDP-glucose (UDP-Glc), UDP-N-acetylglucosamine (UDP-G1cNac), UDP-N-acetyl-galactosamine (UDP-GalNac) and CTP were measured by the ion-pair HPLC technique after perchloric acid extraction of sedimented cells. Very stable values of the adenylate energy charge were found whereas the ratio of trinucleotides of the purine pool to these of the pyrimidine pool and the ratio between UTP and UDP-N-acetyl-sugars changed rapidly during different growth phases.
KeywordsStationary Growth Phase Nucleotide Pool Adenylate Energy Charge Roller Bottle Perchloric Acid Extraction
Unable to display preview. Download preview PDF.
- 1.Atkinson, D.A. Cellular energy metabolism and its regulation (1977). Academic Press New YorkGoogle Scholar
- 6.Lehmann, J.; Vorlop, J.; Büntemeyer, H. (1988) Bubble-free reactors and their development for continuous culture with cell recycle. In: R.E. Spier and J.B. Griffiths (eds.) Animal Cell Biotechnology Vol. 3, pp. 221–237Google Scholar
- 8.Ryll, T.; Jäger, V.; Wagner, R. (1990) Intracellular concentration of ATP and other nucleotides during continuous cultivation of hybridoma cells. In: Production of Biologicals from Animal Cells in Culture Research, Development and Achievements (Spier, R.E.; Griffiths, J.B.;eds.) Butterworths, Kent, UK, in pressGoogle Scholar
- 9.Ryll, T., Wagner, R. (1991) An improved ion-pair HPLC method for the quantification of intracellular bases, nucleosides, nucleotides and sugar-nucleotides in animal cells. J. Chromatogr., submittedGoogle Scholar
- 14.Chapman, A.G.; Fall, L.; Atkinson, D.E. (1971) Adenylate energy charge in Escherichia coli during growth and starvation. Journal Bacteriology 108, 1072–1086Google Scholar
- 15.Marx, U.; Jäger, V.; Kiessig, S.T.; Grunow, R.; von Baehr, R. (1990) Appearance of nonspecific amounts of monoclonal antibodys during fermentation caused by decreased cell viability. In: Production of Biologicals from Animal Cells in Culture Research, Development and Achievements (Spier, R.E.; Griffiths, J.B.; eds.) Butterworths, Kent, UK, in pressGoogle Scholar