Intracellular flux analysis in hybridomas using mass balances and in vitro (13)C nmr

Biotechnology and Bioengineering
C Zupke, G Stephanopoulos

Abstract

Intracellular fluxes are important in defining cellular physiology and its changes in response to environmental variations. Stoichiometric balances combined with extra cellular metabolite measurements were applied to the estimation of intracellular fluxes and the study of energy metabolism in the hybridoma cell line ATCC CRL 1606. Redundant measurements allowed the evaluation of the consistency of the stoichiometry, measurements, and pseudo-steady-state assumption leading to refinement of the assumed biochemistry and identification of measurement errors. To validate the flux estimates, two batch experiments were performed with glucose labeled in the 1 position with (13)C. The distribution of (13)C in secreted lactate was measured via nuclear magnetic resonance spectroscopy (NMR) and compared to that predicted from the estimated intracellular fluxes. There was good agreement between the measured and estimated isotope distributions, demonstrating the validity of the flux estimates obtained from stoichiometric balances. (c) 1995 John Wiley & Sons, Inc.

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Citations

May 5, 1996·Biotechnology and Bioengineering·H P BonariusJ Tramper
Jul 19, 2008·Biotechnology and Bioengineering·K SchmidtJ Villadsen
Jul 19, 2008·Biotechnology and Bioengineering·S M ParkG Stephanopoulos
Jul 9, 1999·Biotechnology and Bioengineering·B D FollstadD I Wang
Nov 27, 2002·Biotechnology and Bioengineering·Julian T KeanePeter P Gray
Oct 20, 1996·Biotechnology and Bioengineering·K B Konstantinov
Dec 1, 1995·Applied Microbiology and Biotechnology·C ZupkeG Stephanopoulos
Sep 18, 2010·Bioprocess and Biosystems Engineering·Olivier HenryAmine Kamen
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Feb 13, 2001·Mathematical Biosciences·W Wiechert, M Wurzel
Apr 25, 2003·Mathematical Biosciences·Nicole Isermann, Wolfgang Wiechert
Sep 14, 1999·Journal of Biotechnology·T W SimpsonG Stephanopoulos
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Apr 21, 1999·Current Opinion in Biotechnology·J Varner, D Ramkrishna
May 30, 2001·Current Opinion in Microbiology·D E Stafford, G Stephanopoulos
Jul 10, 2002·Biomolecular Engineering·Arne BuchholzRalf Takors
Sep 18, 2001·European Journal of Biochemistry·K S YanagimachiG Stephanopoulos
Sep 9, 1999·Tissue Engineering·K LeeM L Yarmush
Oct 20, 2005·Journal of Bioscience and Bioengineering·Chen YangKazuyuki Shimizu
Oct 20, 2005·Journal of Bioscience and Bioengineering·Anshu GambhirWei-Shou Hu
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Sep 1, 2015·Current Opinion in Biotechnology·Maciek R Antoniewicz
Dec 19, 2016·Analytical Biochemistry·Firas S MidaniSantiago Schnell

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