Isolation and characterization of an insect cell line able to perform complex N-linked glycosylation on recombinant proteins

Ogonah, O.W. and Freedman, R.B. and Jenkins, N. and Patel, K. and Rooney, B.C. (1996) Isolation and characterization of an insect cell line able to perform complex N-linked glycosylation on recombinant proteins. Bio-Technology, 14 (2). pp. 197-202. ISSN 0733-222X. (The full text of this publication is not available from this repository)

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Official URL
http://dx.doi.org/10.1038/nbt0296-197

Abstract

Site specific characterization of the N-glycan structures in human interferon gamma (IFN-gamma) derived from baculovirus-infected insect cells was performed using a combination of reverse-phase, high-performance liquid chromatography (rHPLC) and matrix assisted laser desorption time of flight (MALDI-TOF) mass spectrometry, IFN-gamma was produced in two cell lines, an Estigmena acrea-derived subclone (Ea4), and Spodoptera frugiperda cells (Sf9), Both IFN-gamma N-glycosylation sites (Asn(25) and Asn(97)) were characterized, Site-specific differences were observed in both the percentage of sites occupied by N-linked glycans and the types of structure associated with each site, The glycosylation capabilities and glycan processing of Sf9 were limited to the generation of chitobiose [GlcNAc(2)], truncated tri-mannose core [Man(3)GlcNAc(2)], or oligomannose structures, The glycosylation abilities of Ea4 cells were more extensive, producing IFN-gamma molecules incorporating oligosaccharides with GlcNAc and Gal residues on the outer arms (hybrid or complex type N-glycans), as well as oligomannose N-glycans, Incorporation of an alpha 1-6 linked fucose residue (<70% in Sf9 and <88% in Ea4) was confined to the Asn(25) glycosylation site. These findings demonstrate the more extensive N-glycosylation capabilities of the E(1) acrea-derived Ea4, compared to current insect cell lines used for the expression of recombinant proteins.

Item Type: Article
Subjects: Q Science > QR Microbiology
Divisions: Faculties > Science Technology and Medical Studies > School of Biosciences
Depositing User: F.D. Zabet
Date Deposited: 10 Jun 2009 18:52
Last Modified: 10 Jun 2009 18:52
Resource URI: http://kar.kent.ac.uk/id/eprint/18707 (The current URI for this page, for reference purposes)
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