Quantitative characterization of metabolism and metabolic shifts during growth of the new human cell line AGE1.HN using time resolved metabolic flux analysis

Niklas J, Skerhutt EM, Sandig V, Noll T, Heinzle E (2011)
Bioprocess and Biosystems Engineering 34(5): 533-545.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
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Abstract / Bemerkung
For the improved production of vaccines and therapeutic proteins, a detailed understanding of the metabolic dynamics during batch or fed-batch production is requested. To study the new human cell line AGE1.HN, a flexible metabolic flux analysis method was developed that is considering dynamic changes in growth and metabolism during cultivation. This method comprises analysis of formation of cellular components as well as conversion of major substrates and products, spline fitting of dynamic data and flux estimation using metabolite balancing. During batch cultivation of AGE1.HN three distinct phases were observed, an initial one with consumption of pyruvate and high glycolytic activity, a second characterized by a highly efficient metabolism with very little energy spilling waste production and a third with glutamine limitation and decreasing viability. Main events triggering changes in cellular metabolism were depletion of pyruvate and glutamine. Potential targets for the improvement identified from the analysis are (i) reduction of overflow metabolism in the beginning of cultivation, e.g. accomplished by reduction of pyruvate content in the medium and (ii) prolongation of phase 2 with its highly efficient energy metabolism applying e.g. specific feeding strategies. The method presented allows fast and reliable metabolic flux analysis during the development of producer cells and production processes from microtiter plate to large scale reactors with moderate analytical and computational effort. It seems well suited to guide media optimization and genetic engineering of producing cell lines.
Erscheinungsjahr
Zeitschriftentitel
Bioprocess and Biosystems Engineering
Band
34
Zeitschriftennummer
5
Seite
533-545
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eISSN
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Niklas J, Skerhutt EM, Sandig V, Noll T, Heinzle E. Quantitative characterization of metabolism and metabolic shifts during growth of the new human cell line AGE1.HN using time resolved metabolic flux analysis. Bioprocess and Biosystems Engineering. 2011;34(5):533-545.
Niklas, J., Skerhutt, E. M., Sandig, V., Noll, T., & Heinzle, E. (2011). Quantitative characterization of metabolism and metabolic shifts during growth of the new human cell line AGE1.HN using time resolved metabolic flux analysis. Bioprocess and Biosystems Engineering, 34(5), 533-545. doi:10.1007/s00449-010-0502-y
Niklas, J., Skerhutt, E. M., Sandig, V., Noll, T., and Heinzle, E. (2011). Quantitative characterization of metabolism and metabolic shifts during growth of the new human cell line AGE1.HN using time resolved metabolic flux analysis. Bioprocess and Biosystems Engineering 34, 533-545.
Niklas, J., et al., 2011. Quantitative characterization of metabolism and metabolic shifts during growth of the new human cell line AGE1.HN using time resolved metabolic flux analysis. Bioprocess and Biosystems Engineering, 34(5), p 533-545.
J. Niklas, et al., “Quantitative characterization of metabolism and metabolic shifts during growth of the new human cell line AGE1.HN using time resolved metabolic flux analysis”, Bioprocess and Biosystems Engineering, vol. 34, 2011, pp. 533-545.
Niklas, J., Skerhutt, E.M., Sandig, V., Noll, T., Heinzle, E.: Quantitative characterization of metabolism and metabolic shifts during growth of the new human cell line AGE1.HN using time resolved metabolic flux analysis. Bioprocess and Biosystems Engineering. 34, 533-545 (2011).
Niklas, Jens, Skerhutt, Eva Maria, Sandig, Volker, Noll, Thomas, and Heinzle, Elmar. “Quantitative characterization of metabolism and metabolic shifts during growth of the new human cell line AGE1.HN using time resolved metabolic flux analysis”. Bioprocess and Biosystems Engineering 34.5 (2011): 533-545.

35 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Metabolism and metabolic burden by α1-antitrypsin production in human AGE1.HN cells.
Niklas J, Priesnitz C, Rose T, Sandig V, Heinzle E., Metab Eng 16(), 2013
PMID: 23376655
Metabolic signatures of GS-CHO cell clones associated with butyrate treatment and culture phase transition.
Carinhas N, Duarte TM, Barreiro LC, Carrondo MJ, Alves PM, Teixeira AP., Biotechnol Bioeng 110(12), 2013
PMID: 23794452
Towards dynamic metabolic flux analysis in CHO cell cultures.
Ahn WS, Antoniewicz MR., Biotechnol J 7(1), 2012
PMID: 22102428
Isotope labeling experiments in metabolomics and fluxomics.
Klein S, Heinzle E., Wiley Interdiscip Rev Syst Biol Med 4(3), 2012
PMID: 22447740
Compartmentation and channelling of metabolites in the human cell line AGE1.HN(®).
Niklas J, Sandig V, Heinzle E., BMC Proc 5 Suppl 8(), 2011
PMID: 22373369
Producer vs. parental cell - metabolic changes and burden upon α1-antitrypsin production in AGE1.HN(®).
Niklas J, Priesnitz C, Sandig V, Rose T, Heinzle E., BMC Proc 5 Suppl 8(), 2011
PMID: 22373391

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