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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, Mario Jolicoeur

Article (2016)

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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

Subjects: 1800 Chemical engineering > 1800 Chemical engineering
5300 Molecular biology > 5300 Molecular biology
5300 Molecular biology > 5302 Gene expression
5300 Molecular biology > 5305 Protein manipulation and expression
Department: Department of Chemical Engineering
Research Center: Other
Funders: CRSNG / NSERC, Monoclonal Antibody Network (MabNet)
PolyPublie URL: https://publications.polymtl.ca/3602/
Journal Title: AIMS Bioengineering (vol. 3, no. 3)
Publisher: AIMS Press
DOI: 10.3934/bioeng.2016.3.319
Official URL: https://doi.org/10.3934/bioeng.2016.3.319
Date Deposited: 10 Mar 2020 14:40
Last Modified: 11 Nov 2022 14:31
Cite in APA 7: 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), 319-336. https://doi.org/10.3934/bioeng.2016.3.319

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