Multi-dimensional mapping of pyridylamine-labeled N-linked oligosaccharides by capillary electrophoresis |
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Authors: | Lynn R. Zieske Daotian Fu Shaheer H. Khan Roger A. O'Neill |
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Affiliation: | Applied Biosystems Division, Perkin-Elmer Corporation, 850 Lincoln Centre Drive, Foster City, CA 94404, USA |
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Abstract: | A simple, sensitive and reproducible multi-dimensional capillary electrophoresis (CE) oligosaccharide mapping method is reported. The structures of 20 identified N-linked oligosaccharides have been assigned mapping positions from which co-migrating unknown oligosaccharides can be characterized. The separation protocols developed have been demonstrated to separate both charged and neutral oligosaccharides. One dimension involves electroendosmotic flow-assisted CE in a sodium acetate buffer, pH 4.0. A second dimension involves separation based on borate complexation electrophoresis in a polyethylene glycol-containing buffer. A third dimension developed specifically for neutral oligosaccharides, using a sodium phosphate buffer, pH 2.5, has been shown to resolve neutral species not able to be separated by the other two dimensions. Thus, a three-dimensional map was generated to facilitate structural characterization of these oligosaccharides. |
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