Effect of lipid supplements on the production and glycosylation of recombinant interferon-γ expressed in CHO cells

Jenkins, Nigel; Castro, Paula M. L.; Menon, Sunitha; Ison, A. P.; Bull, Alan T.

doi:10.1007/978-94-011-0257-5_23

Cited by 9 publications

(11 citation statements)

References 27 publications

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“…From a biopharmaceutical point of view, it appears essential to produce high quality, consistent, and efficient recombinant therapeutics throughout the processes. Parameters affecting the protein glycosylation have been widely studied during the last two decades 11–17…”

Section: Introductionmentioning

confidence: 99%

Effect of surfactant pluronic F‐68 on CHO cell growth, metabolism, production, and glycosylation of human recombinant IFN‐γ in mild operating conditions

Clincke

Guédon

Yen

et al. 2010

Biotechnology Progress

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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.

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Section: Introductionmentioning

confidence: 99%

Effect of surfactant pluronic F‐68 on CHO cell growth, metabolism, production, and glycosylation of human recombinant IFN‐γ in mild operating conditions

Clincke

Guédon

Yen

et al. 2010

Biotechnology Progress

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“…Regarding the culture process, a number of parameters have been shown to affect glycosylation, including hostcell line (10)(11)(12)(13)(14), process time and cultivation mode (15), ammonia accumulation (15)(16)(17)(18)(19)(20)(21)(22), glucose depletion (23)(24)(25), glucosamine supplementation (26), growth rate (27), lipid availability (28), and dissolved oxygen (29)(30)(31). Moreover, some factors are known to act in an opposite way on glycoprotein structure depending on the production system used; for example, presence of serum may induce a decrease or an increase in the level of galactosylation of antibodies produced by a mouse hybridoma or by CHO cells (32).…”

Section: Introductionmentioning

confidence: 99%

HPCE Monitoring of the N-Glycosylation Pattern and Sialylation of Murine Erythropoietin Produced by CHO Cells in Batch Processes

et al. 2004

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Using capillary electrophoresis coupled to laser-induced fluorescence (HPCE-LIF), it was possible to profile N-linked oligosaccharides from EPO, including species containing sialic acid, during the course of batch cultures performed either in serum-free or serum-containing medium. Although an unusual high heterogeneity of the N-linked oligosaccharides was observed by both SDS-PAGE and HPCE analysis, the patterns of mEPO glycans after desialylation by mild acid hydrolysis were found to be quite constant over the course of the cultures either with or without serum supplementation. In contrast, when the protein was analyzed by HPCE without acidic desialylation, fingerprints of N-linked oligosaccharides changed with time in serum-containing conditions. This phenomenon appeared to be mainly due to the desialylation of mEPO as a result of a sialidase activity released upon cell lysis. These results demonstrate that though a higher EPO titer was obtained in serum supplemented conditions, sialylation of EPO was severely affected by the presence of serum in the culture medium.

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“…The action of phospholipids and fatty acids on enhancement of growth was found to be membrane associated, while antibody enhancement is independent of Brought to you by | New York University Bobst Library Technical Services Authenticated Download Date | 7/6/15 11:58 AM changes in membrane dynamics [32]. Lipid supplements also improved growth and productivity of recombinant proteins in CHO and NS0 cell lines [33,34].…”

Section: Lipidsmentioning

confidence: 90%

5.1 Physiology and Metabolism of Animal Cells for Production

Tayi¹,

Butler²

2014

Animal Cell Biotechnology

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Effect of lipid supplements on the production and glycosylation of recombinant interferon-γ expressed in CHO cells

Cited by 9 publications

References 27 publications

Effect of surfactant pluronic F‐68 on CHO cell growth, metabolism, production, and glycosylation of human recombinant IFN‐γ in mild operating conditions

Effect of surfactant pluronic F‐68 on CHO cell growth, metabolism, production, and glycosylation of human recombinant IFN‐γ in mild operating conditions

HPCE Monitoring of the N-Glycosylation Pattern and Sialylation of Murine Erythropoietin Produced by CHO Cells in Batch Processes

5.1 Physiology and Metabolism of Animal Cells for Production

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