James, D.C. and Freedman, R.B. and Hoare, M. and Jenkins, N. (1994) High-Resolution Separation of Recombinant Human Interferon-Gamma Glycoforms by Micellar Electrokinetic Capillary Chromatography. Analytical Biochemistry, 222 (2). pp. 315-322. ISSN 0003-2697.
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| Official URL http://dx.doi.org/10.1006/abio.1994.1498 |
Abstract
Recombinant human interferon-gamma (IFN-gamma) glycoform populations produced by Chinese hamster ovary cells have been resolved by micellar electrokinetic capillary chromatography (MECC). Separations were performed in uncoated fused silica capillaries at alkaline pH in the presence of micellar concentrations of the anionic detergent sodium dodecyl sulfate (SDS). Maximum resolution was obtained reproducibly with high-ionic-strength borate/SDS electrophoresis buffer. Under the conditions described, glycoform migration time was inversely related to the amount of carbohydrate associated with the protein. Digestion of IFN-gamma with peptide-N-glycosidase F allowed virtual real-time monitoring of glycosidase digests by capillary electrophoresis. Analysis of other digestions with either neuraminidase or endoglycosidase H (endo H) showed most IFN-gamma glycoforms to be sialylated and a minor proportion of glycoforms to be associated with oligomannose structures. While both bovine pancreas ribonuclease B and horseradish peroxidase glycoforms were separated by this technique, proteins glycosylated at multiple sites such as bovine serum fetuin and human alpha(1)-acid glycoprotein were not well resolved by MECC. (C) 1993. Academic Press, Inc.
| Item Type: | Article |
|---|---|
| Subjects: | Q Science > QP Physiology (Living systems) > QP517 Biochemistry |
| Divisions: | Faculties > Science Technology and Medical Studies > School of Biosciences |
| Depositing User: | P. Ogbuji |
| Date Deposited: | 25 Jun 2009 12:03 |
| Last Modified: | 30 May 2012 10:56 |
| Resource URI: | http://kar.kent.ac.uk/id/eprint/20269 (The current URI for this page, for reference purposes) |
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