Host Organisms: Mammalian Cells

Pfizenmaier, Jennifer

doi:10.1002/9783527807796.ch17

Cited by 2 publications

(4 citation statements)

References 149 publications

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“…Today, mammalian producers are typically derived from tissue cells giving Chinese hamster ovary (CHO) cells an outstanding importance for the production of therapeutic proteins (Pfizenmaier and Takors, 2015 ). It has been estimated that these cells experience p CO2 levels of 41–72 mbar under physiological conditions (Altman and Dittmer, 1971 ).…”

Section: Co 2 Induced Growth Phenotypesmentioning

confidence: 99%

CO2 – Intrinsic Product, Essential Substrate, and Regulatory Trigger of Microbial and Mammalian Production Processes

Blombach

Takors

2015

Front. Bioeng. Biotechnol.

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Carbon dioxide formation mirrors the final carbon oxidation steps of aerobic metabolism in microbial and mammalian cells. As a consequence, CO2/HCO3− dissociation equilibria arise in fermenters by the growing culture. Anaplerotic reactions make use of the abundant CO2/HCO3− levels for refueling citric acid cycle demands and for enabling oxaloacetate-derived products. At the same time, CO2 is released manifold in metabolic reactions via decarboxylation activity. The levels of extracellular CO2/HCO3− depend on cellular activities and physical constraints such as hydrostatic pressures, aeration, and the efficiency of mixing in large-scale bioreactors. Besides, local CO2/HCO3− levels might also act as metabolic inhibitors or transcriptional effectors triggering regulatory events inside the cells. This review gives an overview about fundamental physicochemical properties of CO2/HCO3− in microbial and mammalian cultures effecting cellular physiology, production processes, metabolic activity, and transcriptional regulation.

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Section: Co 2 Induced Growth Phenotypesmentioning

confidence: 99%

CO2 – Intrinsic Product, Essential Substrate, and Regulatory Trigger of Microbial and Mammalian Production Processes

Blombach

Takors

2015

Front. Bioeng. Biotechnol.

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“…On the other hand, in vivo compartment-specific metabolome data should be available to allow data-driven studies. Nicolae et al and Pfizenmaier & Takors provided evidence for the importance of subcellular stoichiometric models for estimating fluxes in CHO cells [11,12]. Regarding the latter, Matuszczyk et al [13] applied compartment-specific metabolomics in CHO outlining that cytosolic ATP pools are considerably larger than their mitochondrial counterparts.…”

Section: Symbolsmentioning

confidence: 99%

“…The pH was measured with a conventional pH probe (Mettler-Toledo Inc., Columbus, OH, USA) and maintained at 7.1 using 1 M Na 2 CO 3 or CO 2 gassing. Carbon labeling experiments were performed in the same setup using [U- 13 C 6 ]-d-glucose as a carbon tracer with an average isotopic ratio of 25% [U- 12 C 6 ]-and 75% [U- 13 C 6 ]-d-glucose. Experiments were performed as biological duplicates.…”

Section: Cell Culture and Experimental Set-upmentioning

confidence: 99%

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Compartment-specific metabolome labeling enables the identification of subcellular fluxes that may serve as promising metabolic engineering targets in CHO cells

Wijaya

Ulmer

Hundsdorfer

et al. 2021

Bioprocess Biosyst Eng

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Abstract13C labeling data are used to calculate quantitative intracellular flux patterns reflecting in vivo conditions. Given that approaches for compartment-specific metabolomics exist, the benefits they offer compared to conventional non-compartmented 13C flux studies remain to be determined. Using compartment-specific labeling information of IgG1-producing Chinese hamster ovary cells, this study investigated differences of flux patterns exploiting and ignoring metabolic labeling data of cytosol and mitochondria. Although cellular analysis provided good estimates for the majority of intracellular fluxes, half of the mitochondrial transporters, and NADH and ATP balances, severe differences were found for some reactions. Accurate flux estimations of almost all iso-enzymes heavily depended on the sub-cellular labeling information. Furthermore, key discrepancies were found for the mitochondrial carriers vAGC1 (Aspartate/Glutamate antiporter), vDIC (Malate/H+ symporter), and vOGC (α-ketoglutarate/malate antiporter). Special emphasis is given to the flux of cytosolic malic enzyme (vME): it could not be estimated without the compartment-specific malate labeling information. Interesting enough, cytosolic malic enzyme is an important metabolic engineering target for improving cell-specific IgG1 productivity. Hence, compartment-specific 13C labeling analysis serves as prerequisite for related metabolic engineering studies.

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Host Organisms: Mammalian Cells

Cited by 2 publications

References 149 publications

CO2 – Intrinsic Product, Essential Substrate, and Regulatory Trigger of Microbial and Mammalian Production Processes

CO2 – Intrinsic Product, Essential Substrate, and Regulatory Trigger of Microbial and Mammalian Production Processes

Compartment-specific metabolome labeling enables the identification of subcellular fluxes that may serve as promising metabolic engineering targets in CHO cells

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