• Home
  • Search Results
  • Intracellular flux analysis applied to the effect of dissolved oxygen on hybridomas.

Intracellular flux analysis applied to the effect of dissolved oxygen on hybridomas.

Applied microbiology and biotechnology (1995-12-01)
C Zupke, A J Sinskey, G Stephanopoulos
ABSTRACT

Quantitative estimates of intracellular fluxes and measurements of intracellular concentrations were used to evaluate the effect of dissolved oxygen (DO) concentration on CRL 1606 hybridoma cells in batch culture. The estimates of intracellular fluxes were generated by combining material balances with measurements of extracellular metabolite rates of change. Experiments were performed at DO levels of 60% and 1% air saturation, as well as under oxygen-limited conditions. Cell extracts were analyzed to evaluate the effect of DO on the intracellular concentrations of the glutamate dehydrogenase reactants, as well as the redox state of the pyridine nucleotides in the cytosol and mitochondria. The relationship between cell density and pyridine nucleotide redox state was also investigated. Dissolved oxygen concentration had a significant effect on nitrogen metabolism and the flux through glutamate dehydrogenase was found to reverse at low DO, favoring glutamate formation. The NAD in the cytosol and mitochondria was more reduced under low DO conditions while the cytosolic NAD was more oxidized at low DO. Cytosolic NAD was reduced at higher cell densities while the redox states of cytosolic NADP and mitochondrial NAD did not exhibit significant variation with cell density. These results point to the fundamental role of the intracellular oxidation/reduction state in cell physiology and the possibility of controlling physiological processes through modulation of the dissolved oxygen level or the oxidation/reduction potential of the culture.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ammonia solution, 7 N in methanol
Sigma-Aldrich
Ammonia, anhydrous, ≥99.98%
Sigma-Aldrich
L-Glutamic acid, ReagentPlus®, ≥99% (HPLC)
Sigma-Aldrich
L-(+)-Lactic acid, ≥98%
Sigma-Aldrich
Pyruvic acid, 98%
Sigma-Aldrich
α-Ketoglutaric acid, 99.0-101.0% (T)
Sigma-Aldrich
L-Glutamic acid, from non-animal source, meets EP testing specifications, suitable for cell culture, 98.5-100.5%
Sigma-Aldrich
3-Hydroxybutyric acid, 95%
Sigma-Aldrich
Oxaloacetic acid, ≥97% (HPLC)
Sigma-Aldrich
Ammonia solution, 2.0 M in ethanol
Sigma-Aldrich
L-(−)-Malic acid, ≥95% (titration)
Sigma-Aldrich
Ammonia solution, 0.4 M in THF
Sigma-Aldrich
L-(+)-Lactic acid, BioXtra, ≥98% (titration)
Sigma-Aldrich
Ammonia solution, 0.4 M in dioxane
Supelco
L-(+)-Lactic acid, analytical standard
Sigma-Aldrich
α-Ketoglutaric acid, BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
L-Glutamic acid, BioUltra, ≥99.5% (NT)
Sigma-Aldrich
L-(−)-Malic acid, ReagentPlus®, ≥99%
Sigma-Aldrich
L-(−)-Malic acid, 97%, optical purity ee: 99% (GLC)
Sigma-Aldrich
Ammonia solution, 4 M in methanol
Supelco
L-Glutamic acid, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
L-(+)-Lactic acid, 80%
Sigma-Aldrich
Ammonia solution, 2.0 M in isopropanol
Sigma-Aldrich
L-(+)-Lactic acid solution, in H2O, ≥85%
Sigma-Aldrich
Ammonia solution, 2.0 M in methanol
Sigma-Aldrich
L-(+)-Lactic acid solution, meets analytical specification of Ph. Eur., BP, 88-92% total acid basis
Sigma-Aldrich
L-(−)-Malic acid, BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Pyruvic acid, ≥97%, FG
Sigma-Aldrich
Oxaloacetic acid, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥97% (HPLC)
Sigma-Aldrich
L-Glutamic acid, FCC

Social Media

LinkedIn icon
Twitter icon
Facebook Icon
Instagram Icon

MilliporeSigma

Research. Development. Production.

We are a leading supplier to the global Life Science industry with solutions and services for research, biotechnology development and production, and pharmaceutical drug therapy development and production.

© 2021 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved.

Reproduction of any materials from the site is strictly forbidden without permission.