Multi-dimensional mapping of pyridylamine-labeled N-linked oligosaccharides by capillary electrophoresis.
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.
Zieske
L R
LR
Applied Biosystems Division, Perkin-Elmer Corporation, Foster City, CA 94404, USA.
Fu
D
D
Khan
S H
SH
O'Neill
R A
RA
D000631:Aminopyridines / Q000737:chemistry*; D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002417:Cattle; D019075:Electrophoresis, Capillary; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry; D012794:Sialic Acids / Q000032:analysis / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013431:Sulfates / Q000032:analysis; D013552:Swine
D000631:Aminopyridines; D005456:Fluorescent Dyes; D009844:Oligosaccharides; D012794:Sialic Acids; D013431:Sulfates; C032439:alpha-aminopyridine