Ogonah, Olotu W. and Freedman, Robert B. and Jenkins, Nigel and Patel, Kirit and Rooney, Barrie 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 currently available from this repository. You may be able to access a copy if URLs are provided)
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.
|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:||05 Jun 2014 11:59|
|Resource URI:||https://kar.kent.ac.uk/id/eprint/18707 (The current URI for this page, for reference purposes)|