A multi-pronged investigation into the effect of glucose starvation and culture duration on fed-batch CHO cell culture

• Published in: Biotechnology and bioengineering 2015-10, Vol.112 (10), p.2172-2184
• Main Authors: Fan, Yuzhou, Jimenez Del Val, Ioscani, Müller, Christian, Lund, Anne Mathilde, Sen, Jette Wagtberg, Rasmussen, Søren Kofoed, Kontoravdi, Cleo, Baycin-Hizal, Deniz, Betenbaugh, Michael J., Weilguny, Dietmar, Andersen, Mikael Rørdam
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - biosynthesis; Antibodies, Monoclonal - genetics; Biotechnology; Cell culture; Cell Culture Techniques - methods; chinese hamster ovary cells; CHO Cells; Correlation; Cricetulus; Culture; culture duration; Culture Media - chemistry; fed-batch; Glucose; Glucose - metabolism; glucose starvation; Glycosylation; Monoclonal antibodies; monoclonal antibody; Productivity; Proteins; Proteomics; Recombinant Proteins - genetics; Recombinant Proteins - metabolism
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.25620

ABSTRACT In this study, omics‐based analysis tools were used to explore the effect of glucose starvation and culture duration on monoclonal antibody (mAb) production in fed‐batch CHO cell culture to gain better insight into how these parameters can be controlled to ensure optimal mAb productivity and quality. Titer and N‐glycosylation of mAbs, as well as proteomic signature and metabolic status of the production cells in the culture were assessed. We found that the impact of glucose starvation on the titer and N‐glycosylation of mAbs was dependent on the degree of starvation during early stationary phase of the fed‐batch culture. Higher degree of glucose starvation reduced intracellular concentrations of UDP‐GlcNAc and UDP‐GalNAc, but increased the levels of UDP‐Glc and UDP‐Gal. Increased GlcNAc and Gal occupancy correlated well with increased degree of glucose starvation, which can be attributed to the interplay between the dilution effect associated with change in specific productivity of mAbs and the changed nucleotide sugar metabolism. Herein, we also show and discuss that increased cell culture duration negatively affect the maturation of glycans. In addition, comparative proteomics analysis of cells was conducted to observe differences in protein abundance between early growth and early stationary phases. Generally higher expression of proteins involved in regulating cellular metabolism, extracellular matrix, apoptosis, protein secretion and glycosylation was found in early stationary phase. These analyses offered a systematic view of the intrinsic properties of these cells and allowed us to explore the root causes correlating culture duration with variations in the productivity and glycosylation quality of monoclonal antibodies produced with CHO cells. Biotechnol. Bioeng. 2015;112: 2172–2184. © 2015 Wiley Periodicals, Inc. In this study, a multi‐pronged approach has been used to investigate the effect of glucose starvation and culture duration on mAb production in fedbatch CHO cell culture. Mechanistic insights of the effect are provided by systematic analysis of culture behaviour, mAb titer, N‐glycosylation and proteomic and metabolic status of the cells.