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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Abstract
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.
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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.
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.
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Data provided by Europe PubMed Central.

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Insights into pH-induced metabolic switch by flux balance analysis.
Ivarsson M, Noh H, Morbidelli M, Soos M., Biotechnol. Prog. 31(2), 2015
PMID: 25906421
Methods and advances in metabolic flux analysis: a mini-review.
Antoniewicz MR., J. Ind. Microbiol. Biotechnol. 42(3), 2015
PMID: 25613286
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Zhang Z, Shen T, Rui B, Zhou W, Zhou X, Shang C, Xin C, Liu X, Li G, Jiang J, Li C, Li R, Han M, You S, Yu G, Yi Y, Wen H, Liu Z, Xie X., Nucleic Acids Res. 43(Database issue), 2015
PMID: 25392417
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Rehberg M, Ritter JB, Reichl U., PLoS Comput. Biol. 10(10), 2014
PMID: 25329309
The avian cell line AGE1.CR.pIX characterized by metabolic flux analysis.
Lohr V, Hadicke O, Genzel Y, Jordan I, Buntemeyer H, Klamt S, Reichl U., BMC Biotechnol. 14(), 2014
PMID: 25077436
The influence of cell growth and enzyme activity changes on intracellular metabolite dynamics in AGE1.HN.AAT cells.
Rath AG, Rehberg M, Janke R, Genzel Y, Scholz S, Noll T, Rose T, Sandig V, Reichl U., J. Biotechnol. 178(), 2014
PMID: 24657347
Molecular physiology of the dynamic regulation of carbon catabolite repression in Escherichia coli.
Borirak O, Bekker M, Hellingwerf KJ., Microbiology (Reading, Engl.) 160(Pt 6), 2014
PMID: 24603062
Dynamics of growth and metabolism controlled by glutamine availability in Chinese hamster ovary cells.
Wahrheit J, Nicolae A, Heinzle E., Appl. Microbiol. Biotechnol. 98(4), 2014
PMID: 24362913
Changes in intracellular metabolite pools during growth of adherent MDCK cells in two different media.
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PMID: 24169951
Expression of recombinant antibodies.
Frenzel A, Hust M, Schirrmann T., Front Immunol 4(), 2013
PMID: 23908655
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
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
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Isotope labeling experiments in metabolomics and fluxomics.
Klein S, Heinzle E., Wiley Interdiscip Rev Syst Biol Med 4(3), 2012
PMID: 22447740
Producer vs. parental cell - metabolic changes and burden upon α1-antitrypsin production in AGE1.HN(®).
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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

60 References

Data provided by Europe PubMed Central.

Engineering of a mammalian cell line for reduction of lactate formation and high monoclonal antibody production.
Chen K, Liu Q, Xie L, Sharp PA, Wang DI., Biotechnol. Bioeng. 72(1), 2001
PMID: 11084594
The energy spilling reactions of bacteria and other organisms.
Russell JB., J. Mol. Microbiol. Biotechnol. 13(1-3), 2007
PMID: 17693707
Metabolic flux analysis in eukaryotes.
Niklas J, Schneider K, Heinzle E., Curr. Opin. Biotechnol. 21(1), 2010
PMID: 20163950
Metabolic flux analysis in mammalian cell culture.
Quek LE, Dietmair S, Kromer JO, Nielsen LK., Metab. Eng. 12(2), 2010
PMID: 19833223
Intracellular flux analysis in hybridomas using mass balances and in vitro (13)C nmr.
Zupke C, Stephanopoulos G., Biotechnol. Bioeng. 45(4), 1995
PMID: 18623182
Metabolite-balancing techniques vs. 13C tracer experiments to determine metabolic fluxes in hybridoma cells.
Bonarius HP, Timmerarends B, de Gooijer CD, Tramper J., Biotechnol. Bioeng. 58(2-3), 1998
PMID: 10191398
Mass isotopomer study of the nonoxidative pathways of the pentose cycle with [1,2-13C2]glucose.
Lee WN, Boros LG, Puigjaner J, Bassilian S, Lim S, Cascante M., Am. J. Physiol. 274(5 Pt 1), 1998
PMID: 9612242

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