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Overexpression of G6PDH does not affect the behavior of HEK-293 clones stably expressing interferon-α2b

Edwige Arnold, Iness Hammami, Jingkui Chen, Sachin Gupte, Yves Durocher and Mario Jolicoeur

Article (2016)

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Cite this document: Arnold, E., Hammami, I., Chen, J., Gupte, S., Durocher, Y. & Jolicoeur, M. (2016). Overexpression of G6PDH does not affect the behavior of HEK-293 clones stably expressing interferon-α2b. AIMS Bioengineering, 3(3), p. 319-336. doi:10.3934/bioeng.2016.3.319
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Abstract

HEK293 cells are gaining in interest for the production of recombinant proteins. However, further understanding and engineering of cell metabolism are still needed to improve protein titers. The importance of G6PDH has already been studied regarding redox balance, and a correlation has recently been established between the oxidative pentose phosphate pathway (PPP) and antibody peak production. In this work, HEK293 cells stably expressing interferon-α2b, a parental clone, and a further engineered clone expressing the cytosolic yeast pyruvate-carboxylase (PYC) were transiently transfected to overexpress glucose-6-phosphate dehydrogenase (G6PDH) and increase fluxes through the PPP. The aim of the study was thus to evaluate the effect of overexpressing G6PDH on the “pull” effect brought by the PYC phenotype. Results indicate that the cell metabolism is, however, highly robust, showing a highly regulated PPP damping the potential effects of overexpressing G6PDH. A metabolomic study also clearly demonstrates, by metabolites profiling, that the PYC clone has a highly robust and more efficient metabolic efficiency, compared to its parental clone.

Uncontrolled Keywords

G6PDH overexpression; Pentose Phosphate Pathway; HEK-293; metabolomics; cell energetics; Pyruvate Carboxylase

Open Access document in PolyPublie
Subjects: 1800 Génie chimique > 1800 Génie chimique
5300 Biologie moléculaire > 5300 Biologie moléculaire
5300 Biologie moléculaire > 5302 Expression génétique
5300 Biologie moléculaire > 5305 Expression et recombinaison protéiques
Department: Département de génie chimique
Research Center: Autre
Funders: CRSNG / NSERC, Monoclonal Antibody Network (MabNet)
Date Deposited: 10 Mar 2020 14:40
Last Modified: 25 Mar 2020 14:40
PolyPublie URL: https://publications.polymtl.ca/3602/
Document issued by the official publisher
Journal Title: AIMS Bioengineering (vol. 3, no. 3)
Publisher: AIMS Press
Official URL: https://doi.org/10.3934/bioeng.2016.3.319

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