The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody
| dc.contributor.author | Liu, Bo | |
| dc.contributor.examiningcommittee | Brassinga,Ann Karen(Microbiology) Schweizer,Frank(Microbiology) | en_US |
| dc.contributor.supervisor | Butler, Michael (Microbiology) | en_US |
| dc.date.accessioned | 2014-10-28T12:28:57Z | |
| dc.date.available | 2014-10-28T12:28:57Z | |
| dc.date.issued | 2014 | en_US |
| dc.degree.discipline | Microbiology | en_US |
| dc.degree.level | Master of Science (M.Sc.) | en_US |
| dc.description.abstract | The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25 mM) of cultures established at high density (>106 cells/ml) over 24 h. The resulting proportion of non-glycosylated Mab was directly correlated to the exposure time of cells to media depleted of glucose. Deprivation of glucose for the full 24 h resulted in a 52% non-glycosylated Mab fraction. Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI), which showed a direct correlation to the cell specific glucose uptake. These findings are important in relation to the low substrate that may occur in fed-batch cultures for Mab production. | en_US |
| dc.description.note | February 2015 | en_US |
| dc.identifier.citation | Liu, B., Spearman, M., Doering, J., Lattová, E., Perreault, H., Butler, M., 2014. The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. J biotechnology 170, 17–27. doi:10.1016/j.jbiotec.2013.11.007 | en_US |
| dc.identifier.uri | http://hdl.handle.net/1993/24445 | |
| dc.language.iso | eng | en_US |
| dc.publisher | Journal of Biotechnology | en_US |
| dc.rights | open access | en_US |
| dc.subject | Glycosylation | en_US |
| dc.subject | Monoclonal antibody | en_US |
| dc.subject | CHO cells | en_US |
| dc.subject | Lipid-linked oligosaccharide | en_US |
| dc.title | The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody | en_US |
| dc.type | master thesis | en_US |