title: Effect of hydrocortisone on the production and glycosylation of an Fc-fusion protein in CHO cell cultures.
abstract: Glucocorticoids are known to modulate various cellular functions such as cell proliferation, metabolism, glycosylation, and secretion of many proteins. We tested the effect of hydrocortisone (HC) on cell growth, viability, metabolism, protein production, and glycosylation of an Fc-protein expressing Chinese hamster ovary (CHO) cell culture. HC extended cell viability but impaired cell growth. The inhibitory effect on cell growth was dose-dependent and decreased when the glucocorticoid addition was delayed. When HC was added after 2 or 3 days of culture, an increase in glutamate consumption was observed, which was reversed by the glucocorticoid receptor antagonist mifepristone (Mif). Titer and specific productivity increased in the presence of HC. The increase in titer was only slightly reversed by Mif. On the other hand, Mif by itself induced an increase in titer to a level comparable to or higher than HC. Protein glycosylation was altered by the glucocorticoid in a dose- and time-dependent manner, with a shift to more acidic bands, which correlated with an increase in sialic acid moieties. This increase, which was not linked to a decrease in extracellular sialidase activity in HC-treated cultures, was reversed by Mif. Predictive models based on design of experiments enabled the definition of optimal conditions for process performance in terms of viability and titer and for the quality of the Fc-fusion protein in terms of glycosylation. The data obtained suggest a use of glucocorticoids for commercial production of Fc-fusion proteins expressed in CHO cells.
authors.affiliation: Merck Serono SA, Corsier-sur-Vevey, Biotech Process Sciences, ZI B, CH-1809 Fenil-sur-Corsier, Switzerland. yolande.rouiller@merckgroup.com
authors.forename: Yolande
authors.initials: Y
authors.lastname: Rouiller
authors.forename: Arnaud
authors.initials: A
authors.lastname: Périlleux
authors.forename: Milène
authors.initials: M
authors.lastname: Marsaut
authors.forename: Matthieu
authors.initials: M
authors.lastname: Stettler
authors.forename: Marie-Noëlle
authors.initials: MN
authors.lastname: Vesin
authors.forename: Hervé
authors.initials: H
authors.lastname: Broly
mesh_terms: D000818:Animals; D016466:CHO Cells; D002455:Cell Division / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D019075:Electrophoresis, Capillary; D006031:Glycosylation; D006854:Hydrocortisone / Q000494:pharmacology*; D015735:Mifepristone / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism*; D020349:Surface Plasmon Resonance
chemical_list: D011993:Recombinant Fusion Proteins; D015735:Mifepristone; D006854:Hydrocortisone