Influence of intracellular nucleotide and nucleotide sugar contents on recombinant interferon-[gamma] glycosylation during batch and fed-batch cultures of CHO cells

• Published in: Biotechnology and bioengineering Vol. 100; no. 4; p. 721
• Main Authors: Kochanowski, N, Blanchard, F, Cacan, R, Chirat, F, Guedon, E, Marc, A, Goergen, J-L
• Format: Journal Article
• Language: English
• Published: New York Wiley Subscription Services, Inc 01.07.2008
• Subjects: Cell culture; Glycosylation; Interferon; Sugar
• ISSN: 0006-3592; 1097-0290

Both the macroheterogeneity of recombinant human IFN-... produced by CHO cells and intracellular levels of nucleotides and sugar nucleotides, have been characterized during batch and fed-batch cultures carried out in different media. Whereas PF-BDM medium was capable to maintain a high percentage of the doubly-glycosylated glycoforms all over the process, mono-glycosylated and non-glycosylated forms increased during the batch culture using SF-RPMI medium. Intracellular level of UTP was higher in PF-BDM all over the batch culture compared to the SF-RPMI process. UDP-Gal accumulated only during the culture performed in PF-BDM medium, probably as a consequence of the reduced UDP-Glc synthesis flux in SF-RPMI medium. When the recombinant CHO cells were cultivated in fed-batch mode, the UTP level remained at a relatively high value in serum-containing RPMI and its titer increased during the fed-phase indicating an excess of biosynthesis. Besides, an accumulation of UDP-Gal occurred as well. Those results all together indicate that UTP and UDP-Glc syntheses in CHO cells cultivated in SF-RPMI medium in batch process, could be limiting during the glycosylation processes of the recombinant IFN-... At last, the determination of the energetic status of the cells over the three studied processes suggested that a relationship between the adenylate energy charge and the glycosylation macroheterogeneity of the recombinant IFN-... may exist. (ProQuest: ... denotes formulae/symbols omitted.)