Multi-dimensional mapping of pyridylamine-labeled N-linked oligosaccharides by capillary electrophoresis

• Published in: Journal of Chromatography A Vol. 720; no. 1; pp. 395 - 407
• Main Authors: Zieske, Lynn R, Fu, Daotian, Khan, Shaheer H, O'Neill, Roger A
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 12-01-1996
• Subjects: Aminopyridines - chemistry; Animals; Carbohydrate Conformation; Carbohydrate Sequence; Cattle; Electrophoresis, Capillary; Fluorescent Dyes - chemistry; Humans; Magnetic Resonance Spectroscopy; Molecular Sequence Data; Oligosaccharides - analysis; Oligosaccharides - chemistry; Sialic Acids - analysis; Sialic Acids - chemistry; Spectrometry, Fluorescence; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Sulfates - analysis; Swine
• ISSN: 0021-9673
• DOI: 10.1016/0021-9673(95)00346-0

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.