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Intracellular flux analysis in hybridomas using mass balances and in vitro 13 C nmr
Author(s) -
Zupke Craig,
Stephanopoulos Gregory
Publication year - 1995
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260450403
Subject(s) - intracellular , metabolic flux analysis , flux (metallurgy) , stoichiometry , chemistry , isotope , metabolite , nuclear magnetic resonance spectroscopy , analytical chemistry (journal) , metabolism , biophysics , biochemistry , chromatography , biology , stereochemistry , physics , nuclear physics , organic chemistry
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. © 1995 John Wiley & Sons, Inc.