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Metabolic engineering of CHO cells to alter lactate metabolism during fed-batch cultures

Cécile Toussaint, Olivier Henry and Yves Durocher

Article (2016)

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Cite this document: Toussaint, C., Henry, O. & Durocher, Y. (2016). Metabolic engineering of CHO cells to alter lactate metabolism during fed-batch cultures. Journal of Biotechnology, 217(C), p. 122-131. doi:10.1016/j.jbiotec.2015.11.010
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Abstract

Recombinant yeast pyruvate carboxylase (PYC2) expression was previously shown to be an effective metabolic engineering strategy for reducing lactate formation in a number of relevant mammalian cell lines, but, in the case of CHO cells, did not consistently lead to significant improvement in terms of cell growth, product titer and energy metabolism efficiency. In the present study, we report on the establishment of a PYC2-expressing CHO cell line producing a monoclonal antibody and displaying a significantly altered lactate metabolism compared to its parental line. All clones exhibiting strong PYC2 expression were shown to experience a significant and systematic metabolic shift toward lactate consumption, as well as a prolonged exponential growth phase leading to an increased maximum cell concentration and volumetric product titer. Of salient interest, PYC2-expressing CHO cells were shown to maintain a highly efficient metabolism in fed-batch cultures, even when exposed to high glucose levels, thereby alleviating the need of controlling nutrient at low levels and the potential negative impact of such strategy on product glycosylation. In bioreactor operated in fed-batch mode, the higher maximum cell density achieved with the PYC2 clone led to a net gain (20%) in final volumetric productivity. (C) 2015 Published by Elsevier B.V.

Uncontrolled Keywords

CHO cells; Fed-batch; Metabolic engineering; Monoclonal antibody; Pyruvate carboxylase; Animals; Antibodies, Monoclonal; Batch Cell Culture Techniques; Bioreactors; CHO Cells; Cricetinae; Cricetulus; Glucose; Glycosylation; Lactic Acid; Metabolic Engineering; Pyruvate Carboxylase; Recombinant Proteins; Antibodies, Monoclonal; Recombinant Proteins; Lactic Acid; Pyruvate Carboxylase; Glucose

Open Access document in PolyPublie
Subjects: 1800 Génie chimique > 1800 Génie chimique
Department: Département de génie chimique
Research Center: Non applicable
Funders: CRSNG / NSERC
Date Deposited: 25 Oct 2018 16:25
Last Modified: 26 Oct 2018 01:20
PolyPublie URL: https://publications.polymtl.ca/3496/
Document issued by the official publisher
Journal Title: Journal of Biotechnology (vol. 217, no. C)
Publisher: Elsevier
Official URL: https://doi.org/10.1016/j.jbiotec.2015.11.010

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