Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 110420
Title Metabolic-flux analysis of continuously cultured hybridoma cells using 13CO2 mass spectrometry in combination with 13C-lactate nuclear magnetic resonance spectroscopy and metabolic balancing
Author(s) Bonarius, H.P.J.; Ozemre, A.; Timmerarends, B.; Skrabal, P.; Tramper, J.; Schmid, G.; Heinzle, E.
Source Biotechnology and Bioengineering 74 (2001)6. - ISSN 0006-3592 - p. 528 - 538.
DOI https://doi.org/10.1002/bit.1145
Department(s) Sub-department of Food and Bioprocess Engineering
Publication type Refereed Article in a scientific journal
Publication year 2001
Abstract Protein production of mammalian-cell culture is limited due to accumulation of waste products such as lactate, CO2, and ammonia. In this study, the intracellular fluxes of hybridoma cells are measured to determine the amount by which various metabolic pathways contribute to the secretion of waste products derived from glucose. Continuously cultured hybridoma cells are grown in medium containing either 1-13C-, 2-13C-, or 6-13C-glucose. The uptake and production rates of amino acids, glucose, ammonia, O2, and CO2 as well as the cellular composition are measured. In addition, the 13C distribution of the lactate produced and alanine produced by the hybridomas is determined by 1H-NMR spectroscopy, and the 13CO2/12CO2 ratio is measured by on-line mass spectrometry. These data are used to calculate the intracellular fluxes of the glycolysis, the pentose phosphate pathway, the TCA cycle, and fluxes involved in amino acid metabolism. It is shown that: (i) approximately 20␘f the glucose consumed is channeled through the pentose shunt; (ii) the glycolysis pathway contributes the most to lactate production, and most of the CO2 is produced by the TCA cycle; (iii) the pyruvate-carboxylase flux is negligibly small; and (iv) the malic-enzyme flux is estimated to be 10␘f the glucose uptake rate. Based on these flux data suggestions are made to engineer a more efficient glucose metabolism in mammalian cells
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