Dynamic Metabolic Flux Analysis Using B-splines to Study the Effects of Temperature Shift on CHO Cell Metabolism

Overview

Journal Metab Eng Commun

Specialty Biochemistry

Date 2021 Jun 21

PMID 34150508

Citations 26

Authors

Veronica S Martinez

Maria Buchsteiner

Peter Gray

Lars K Nielsen

Lake-Ee Quek

Affiliations

Soon will be listed here.

Abstract

Metabolic flux analysis (MFA) is widely used to estimate intracellular fluxes. Conventional MFA, however, is limited to continuous cultures and the mid-exponential growth phase of batch cultures. Dynamic MFA (DMFA) has emerged to characterize time-resolved metabolic fluxes for the entire culture period. Here, the linear DMFA approach was extended using B-spline fitting (B-DMFA) to estimate mass balanced fluxes. Smoother fits were achieved using reduced number of knots and parameters. Additionally, computation time was greatly reduced using a new heuristic algorithm for knot placement. B-DMFA revealed that Chinese hamster ovary cells shifted from 37 °C to 32 °C maintained a constant IgG volume-specific productivity, whereas the productivity for the controls peaked during mid-exponential growth phase and declined afterward. The observed 42% increase in product titer at 32 °C was explained by a prolonged cell growth with high cell viability, a larger cell volume and a more stable volume-specific productivity.

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References

Matasci M, Hacker D, Baldi L, Wurm F . Recombinant therapeutic protein production in cultivated mammalian cells: current status and future prospects. Drug Discov Today Technol. 2014; 5(2-3):e37-42. DOI: 10.1016/j.ddtec.2008.12.003. View

Altamirano C, Illanes A, Becerra S, Cairo J, Godia F . Considerations on the lactate consumption by CHO cells in the presence of galactose. J Biotechnol. 2006; 125(4):547-56. DOI: 10.1016/j.jbiotec.2006.03.023. View

Young J . Metabolic flux rewiring in mammalian cell cultures. Curr Opin Biotechnol. 2013; 24(6):1108-15. PMC: 3775942. DOI: 10.1016/j.copbio.2013.04.016. View

Mahadevan R, Edwards J, Doyle 3rd F . Dynamic flux balance analysis of diauxic growth in Escherichia coli. Biophys J. 2002; 83(3):1331-40. PMC: 1302231. DOI: 10.1016/S0006-3495(02)73903-9. View

Dietmair S, Timmins N, Gray P, Nielsen L, Kromer J . Towards quantitative metabolomics of mammalian cells: development of a metabolite extraction protocol. Anal Biochem. 2010; 404(2):155-64. DOI: 10.1016/j.ab.2010.04.031. View

Nielsen L, Reid S, Greenfield P . Cell cycle model to describe animal cell size variation and lag between cell number and biomass dynamics. Biotechnol Bioeng. 2008; 56(4):372-9. DOI: 10.1002/(SICI)1097-0290(19971120)56:4<372::AID-BIT3>3.0.CO;2-L. View

Vercammen D, Logist F, Van Impe J . Dynamic estimation of specific fluxes in metabolic networks using non-linear dynamic optimization. BMC Syst Biol. 2014; 8:132. PMC: 4280005. DOI: 10.1186/s12918-014-0132-0. View

Martinez V, Gerdtzen Z, Andrews B, Asenjo J . Viral vectors for the treatment of alcoholism: use of metabolic flux analysis for cell cultivation and vector production. Metab Eng. 2009; 12(2):129-37. DOI: 10.1016/j.ymben.2009.09.003. View

Bonarius H, Hatzimanikatis V, Meesters K, De Gooijer C, Schmid G, Tramper J . Metabolic flux analysis of hybridoma cells in different culture media using mass balances. Biotechnol Bioeng. 1996; 50(3):299-318. DOI: 10.1002/(SICI)1097-0290(19960505)50:3<299::AID-BIT9>3.0.CO;2-B. View

10.

Carvalhal A, Marcelino I, Carrondo M . Metabolic changes during cell growth inhibition by p27 overexpression. Appl Microbiol Biotechnol. 2003; 63(2):164-73. DOI: 10.1007/s00253-003-1385-5. View

11.

Vallino J, Stephanopoulos G . Metabolic flux distributions in Corynebacterium glutamicum during growth and lysine overproduction. Biotechnol Bioeng. 1993; 41(6):633-46. DOI: 10.1002/bit.260410606. View

12.

Fox S, Patel U, Yap M, Wang D . Maximizing interferon-gamma production by Chinese hamster ovary cells through temperature shift optimization: experimental and modeling. Biotechnol Bioeng. 2004; 85(2):177-84. DOI: 10.1002/bit.10861. View

13.

Ahn W, Antoniewicz M . Metabolic flux analysis of CHO cells at growth and non-growth phases using isotopic tracers and mass spectrometry. Metab Eng. 2011; 13(5):598-609. DOI: 10.1016/j.ymben.2011.07.002. View

14.

Quek L, Dietmair S, Kromer J, Nielsen L . Metabolic flux analysis in mammalian cell culture. Metab Eng. 2009; 12(2):161-71. DOI: 10.1016/j.ymben.2009.09.002. View

15.

Boghigian B, Seth G, Kiss R, Pfeifer B . Metabolic flux analysis and pharmaceutical production. Metab Eng. 2009; 12(2):81-95. DOI: 10.1016/j.ymben.2009.10.004. View

16.

Niklas J, Schrader E, 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 Biosyst Eng. 2010; 34(5):533-45. PMC: 3092918. DOI: 10.1007/s00449-010-0502-y. View

17.

Quek L, Nielsen L . On the reconstruction of the Mus musculus genome-scale metabolic network model. Genome Inform. 2009; 21:89-100. View

18.

Ahn W, Antoniewicz M . Parallel labeling experiments with [1,2-(13)C]glucose and [U-(13)C]glutamine provide new insights into CHO cell metabolism. Metab Eng. 2012; 15:34-47. DOI: 10.1016/j.ymben.2012.10.001. View

19.

Becker J, Klopprogge C, Zelder O, Heinzle E, Wittmann C . Amplified expression of fructose 1,6-bisphosphatase in Corynebacterium glutamicum increases in vivo flux through the pentose phosphate pathway and lysine production on different carbon sources. Appl Environ Microbiol. 2005; 71(12):8587-96. PMC: 1317465. DOI: 10.1128/AEM.71.12.8587-8596.2005. View

20.

Meadows A, Karnik R, Lam H, Forestell S, Snedecor B . Application of dynamic flux balance analysis to an industrial Escherichia coli fermentation. Metab Eng. 2009; 12(2):150-60. DOI: 10.1016/j.ymben.2009.07.006. View