Hooker, A.D. (1997) Monitoring of recombinant interferon-gamma glycosylation during batch fermentation of CHO cells. Cytotechnology , 23 (1-3). pp. 103-111. ISSN 0920-9069 .
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A Chinese hamster ovary cell line expressing recombinant human interferon-gamma (IFN-gamma) was grown in a 15-l stirred tank fermenter. N-linked carbohydrate populations associated with both Asn(25) and Asn(97) were isolated by reverse-phase HPLC separation of trypsin-digested IFN-gamma and their structure determined by matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-MD) in combination with exoglycosidase array sequencing. The predominant oligosaccharide at both glycosylation sites throughout the culture was a complex biantennary structure, Gal(2)GlcNAc(2)Man(3)GlcNAc(2), which was fucosylated when attached to Asn(25) but not to Asn(97). A gradual decrease in this biantennary structure was observed, with a concomitant increase in the proportion of truncated and high-mannose glycans. These experiments demonstrate the relative stability of glycosylation during batch culture and the definitive site-specific glycosylation data that ran be obtained using MALDI-MS as a monitoring technique.
|Uncontrolled keywords:||batch culture; Chinese hamster ovary; interferon-gamma; mass spectrometry; proteolytic processing; serum-free cultivation|
|Subjects:||Q Science > Q Science (General)|
|Divisions:||Faculties > Science Technology and Medical Studies > School of Biosciences|
|Depositing User:||P. Ogbuji|
|Date Deposited:||29 May 2009 13:20|
|Last Modified:||25 Jun 2012 13:39|
|Resource URI:||http://kar.kent.ac.uk/id/eprint/19602 (The current URI for this page, for reference purposes)|
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