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.

Synthesis, characterization and evaluation of in vitro toxicity in hepatocytes of linear polyesters with varied aromatic and aliphatic co-monomers

“…Hamster Ovary cells, [56] albeit at much higher concentrations than in our assays, through a mechanism that likely involved partial or temporary membrane modification.…”

Rational development of a serum-free medium and fed-batch process for a GS-CHO cell line expressing recombinant antibody

“…Insulin can stimulate cell growth and cell cycle progression, regulate metabolism of glucose and lipid, as well as increase biosynthesis of fatty acids and nucleic acids (Abdeen et al 2011). Pluronic F68, an important medium additive for protecting suspension cells against shear damage, was elaborated in detail for its beneficial effects on CHO cell growth, metabolism, production and glycosylation of human recombinant IFN-c (Clincke et al 2011). Dextran sulfate is an effective dispersing agent for blocking aggregation and inducing single cell suspension under serum-free condition.…”

Myogenic and neurogenic differentiation of human tooth germ stem cells (hTGSCs) are regulated by pluronic block copolymers

“…Each type of poloxamers varies depending on structural characteristics (number of subunits) (Kabanov et al 2002). Pluronic F68 (PEO 76 PPO 29 PEO 76 ) interacts with cellular membranes and prevents mechanical stress caused by agitation (Clincke et al 2011). F68 was shown to exert neuroprotective role against neurotoxic molecules by protecting membrane integrity (Mina et al 2009).…”