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

Cécile Toussaint, Olivier Henry et Yves Durocher

Article de revue (2016)

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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.

Mots clés

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

Sujet(s): 1800 Génie chimique > 1800 Génie chimique
Département: Département de génie chimique
Organismes subventionnaires: CRSNG / NSERC
URL de PolyPublie: https://publications.polymtl.ca/3496/
Titre de la revue: Journal of Biotechnology (vol. 217, no C)
Maison d'édition: Elsevier
DOI: 10.1016/j.jbiotec.2015.11.010
URL officielle: https://doi.org/10.1016/j.jbiotec.2015.11.010
Date du dépôt: 25 oct. 2018 16:25
Dernière modification: 26 sept. 2024 21:46
Citer en APA 7: 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), 122-131. https://doi.org/10.1016/j.jbiotec.2015.11.010

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