N-Glycosylation of Recominant Human Interferon-Gamma Produced in Deferent Animal Expression System

James, D.C. and Freedman, R.B. and Hoare, M. and Ogonah, O.W. and Rooney, B.C. and Larionov, O.A. and Dobrovolsky, V.N. and Lagutin, O.V. and Jenkins, N. (1998) N-Glycosylation of Recominant Human Interferon-Gamma Produced in Deferent Animal Expression System. Biotechnology and Bioengineering , 60 (5). pp. 59-607. ISSN 0006-3592 . (The full text of this publication is not available from this repository)

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Official URL
http://dx.doi.org/10.1002/(SICI)1097-0290(19981205...

Abstract

Recombinant human interferon-gamma (IFN-gamma) was expressed in Chinese hamster ovary cells, baculovirus-infected Sf9 insect cells and the mammary gland of transgenic mice, The N-linked carbohydrate populations associated with both Asn(25), and Asn(97) glycosylation sites were characterized by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) in combination with exoglycosidase array sequencing, a site-specific analysis of dual (2N) and single (1N) site-occupancy variants of IFN-gamma derived from Chinese hamster ovary cells showed that N-glycans were predominantly of the complex bi- and triantennary type, Although Asn(25)-linked glycans were substituted with a core fucose residue, Asn(97) N-glycans were predominantly non-fucosylated, and truncated complex and high-mannose oligosaccharide chains were also evident, Transgenic mouse derived IFN-gamma exhibited considerable site-specific variation in N-glycan structures, Asn(25)-linked carbohydrates were of the complex, core fucosylated type, Asn(97)-linked carbohydrates were mainly of the oligomannose type, with smaller proportions of hybrid and complex N-glycans, Carbohydrates associated with both glycosylation sites of IFN-gamma from Sf9 insect cells were mainly tri-mannosyl core structures, with fucosylation confined to the Asn(25) site, These data demonstrate the profound influence of host cell type and protein structure on the N-glycosylation of recombinant proteins.

Item Type: Article
Uncontrolled keywords: micellar electrokinetic capillary chromatography; capillary isoelectric focusing; Chinese hamster ovary; interferon-gamma; perfusion culture; glycosylation
Subjects: T Technology > T Technology (General)
Divisions: Faculties > Science Technology and Medical Studies > School of Biosciences
Depositing User: P. Ogbuji
Date Deposited: 29 May 2009 09:54
Last Modified: 31 Jul 2012 11:55
Resource URI: http://kar.kent.ac.uk/id/eprint/19617 (The current URI for this page, for reference purposes)
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