Hooker, A.D. and Goldman, M.H. and Markham, N.H. and James, D.C. and Ison, A.P. and Bull, A.T. and Strange, P.G. and Salmon, I. and Baines, A.J. and Jenkins, N. (2007) N-Glycans of Recombinant Human Interferon-Gamma Change During Batch Culture of Chinese - Hamster Ovary Cells. Biotechnology and Bioengineering, 97 (2). 410-424 . ISSN 0006-3592.
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A recombinant Chinese hamster ovary (CHO) cell line making human interferon-gamma (IFN-gamma) was grown in 12-L stirred tank fermenters in three batch fermentations under conditions of constant temperature, pH, and dissolved oxygen tension. In addition to cell growth, metabolite, and productivity data, a detailed analysis of the carbohydrate structures attached to each glycosylation site of IFN-gamma was achieved using matrix-assisted laser desorption mass spectrometry (MALDI-MS) in combination with exoglycosidase array sequencing. Complex biantennary oligosaccharides (particularly Gal(2)GlcNAc(4)Man(3) which was core alpha 1-6 fucosylated at Asn(25) but not at Asn(97)) were most prevalent at both glycosylation sites. However, considerable microheterogeneity arising from the presence of triantennary and truncated glycan structures was also observed. The proportion of the dominant core glycan structure (Gal(2)GlcNAc(4)Man(3) +/- Fuc(1)) decreased by 15-26% during batch culture, with increases in the proportion of oligomannose and truncated glycans over the same time period. Prolonged culture resulting from an extended lag phase led to further accumulation of oligomannose and truncated structures, reaching up to 52% of total glycans attached to Asn(97) by 240 h of culture. The implications of these glycosylation changes for optimizing the time for harvesting cell cultures, and for the clearance of recombinant therapeutic products in vivo are discussed. (C) 1995 John Wiley & Sons, Inc.
|Uncontrolled keywords:||2D PAGE; proteomics; fed-batch culture; partial least squares regression; NS0 murine myeloma; monoclonal antibody|
|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 13:08|
|Last Modified:||11 Jun 2012 14:04|
|Resource URI:||http://kar.kent.ac.uk/id/eprint/19603 (The current URI for this page, for reference purposes)|
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