title: Effect of surfactant pluronic F-68 on CHO cell growth, metabolism, production, and glycosylation of human recombinant IFN-γ in mild operating conditions.
abstract: The control of glycosylation to satisfy regulatory requirements and quality consistency of recombinant proteins produced by different processes has become an important issue. With two N-glycosylation sites, γ-interferon (IFN-γ) can be seen as a prototype of a recombinant therapeutic glycoprotein for this purpose. The effect of the nonionic surfactant Pluronic F-68 (PF-68) on cell growth and death was investigated, as well as production and glycosylation of recombinant IFN-γ produced by a CHO cell line that was maintained in a rich protein-free medium in the absence or presence of low agitation. Under these conditions, a dose-dependent effect of PF-68 (0-0.1%) was shown not only to significantly enhance growth but also to reduce cell lysis. Interestingly, supplementing the culture medium with PF-68 led to increased IFN-γ production as a result of both higher cell densities and a higher specific production rate of IFN-γ. If cells were grown with agitation, lack of PF-68 in the culture medium decreased the fraction of the fully glycosylated IFN-γ glycoform (2N) from 80% to 65-70% during the initial period. This effect appeared to be due to a lag phase in cell growth observed during this period. Finally, a global kinetic study of CHO cell metabolism indicated higher efficiency in the utilization of the two major carbon substrates when cultures were supplemented with PF-68. Therefore, these results highlight the importance of understanding how media surfactant can affect cell growth as well as cell death and the product quality of a recombinant glycoprotein expressed in CHO cell cultures.
authors.affiliation: Laboratoire Réactions et Génie des Procédés, UPR-CNRS 3349, ENSAIA-INPL, Nancy Université, Vandoeuvre-lès-Nancy, France.
authors.forename: Marie-Françoise
authors.initials: MF
authors.lastname: Clincke
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Guedon
authors.forename: Frances T
authors.initials: FT
authors.lastname: Yen
authors.forename: Virginie
authors.initials: V
authors.lastname: Ogier
authors.forename: Olivier
authors.initials: O
authors.lastname: Roitel
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Goergen
mesh_terms: D000818:Animals; D016466:CHO Cells; D002455:Cell Division / Q000187:drug effects*; D006224:Cricetinae; D003412:Cricetulus; D006031:Glycosylation; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism*; D020442:Poloxamer / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D020442:Poloxamer; D007371:Interferon-gamma