Monitoring of recombinant interferon-gamma glycosylation during batch fermentation of CHO cells

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 . (The full text of this publication is not available from this repository)

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Official URL
http://dx.doi.org/10.1023/A:1007947130709

Abstract

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.

Item Type: Article
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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