Impact of dynamic online fed‐batch strategies on metabolism, productivity and N‐glycosylation quality in CHO cell cultures

Citations Over TimeTop 10% of 2004 papers

Abstract

Abstract As we pursue the means to improve yields to meet growing therapy demands, it is important to examine the impact of process control on glycosylation patterns to ensure product efficacy and consistency. In this study, we describe a dynamic on‐line fed‐batch strategy based on low glutamine/glucose concentrations and its impact on cellular metabolism and, more importantly, the productivity and N‐glycosylation quality of a model recombinant glycoprotein, interferon gamma (IFN‐γ). We found that low glutamine fed‐batch strategy enabled up to 10‐fold improvement in IFN‐γ yields, which can be attributed to reduced specific productivity of ammonia and lactate. Furthermore, the low glutamine concentration (0.3 mM) used in this fed‐batch strategy could maintain both the N‐glycosylation macro‐ and microheterogeneity of IFN‐γ. However, very low glutamine (<0.1 mM) or glucose (<0.70 mM) concentrations can lead to decreased sialylation and increased presence of minor glycan species consisting of hybrid and high‐mannose types. This shows that glycan chain extension and sialylation can be affected by nutrient limitation. In addition to nutrient limitation, we also found that N‐glycosylation quality can be detrimentally affected by low culture viability. IFN‐γ purified at low culture viability had both lower sialylation as well as glycans of lower molecular masses, which can be attributed to extensive degradation by intracellular glycosidases released by cytolysis. Therefore, in order to maintain good N‐glycosylation quality, there is a need to consider both culture viability and nutrient control setpoint in a nutrient‐limiting fed‐batch culture strategy. A greater understanding of these major factors that affect N‐glycosylation quality would surely facilitate future development of effective process controls. © 2004 Wiley Periodicals, Inc.

Related Papers

• → A dynamic mathematical model for monoclonal antibody N‐linked glycosylation and nucleotide sugar donor transport within a maturing Golgi apparatus(2011)133 cited
• → Simultaneous characterization of SNPs and N-glycans from multiple glycosylation sites of intact β-2-glycoprotein-1 (B2GP1) by ESI-qTOF-MS(2019)7 cited
• → Mass spectrometry in the characterization of human genetic N‐glycosylation defects(2008)37 cited
• → Protein Glycosylation: The Basic Science(2009)3 cited
• Protuglikanska protutijela i glikozilacija imunoglobulina g u raku debeloga crijeva(2015)