Analysis of carbohydrates in glycoproteins by high-performance liquid chromatography and high-performance capillary electrophoresis.
We describe two methods for the analysis of oligosaccharide chains in glycoproteins by high-performance liquid chromatography (HPLC) and high-performance capillary electrophoresis (HPCE). O- and N-glycosidically linked oligosaccharides released from glycoproteins can be identified as their borohydride-reduced forms by anion-exchange HPLC with pulsed amperometric detection. N-Glycosidically linked oligosaccharides can also be analyzed as 2-aminopyridine derivatives by HPCE in direct zone electrophoresis mode in an acidic phosphate buffer and zone electrophoresis mode as borate complexes in an alkaline buffer. We also present a convenient procedure for the analysis of the constituent monosaccharides of these oligosaccharides chains by HPLC based on reversed-phase partition mode as 1-phenyl-3-methyl-5-pyrazolone derivatives.
Kakehi
K
K
Faculty of Pharmaceutical Sciences, Kinki University, Higashi-Osaka, Japan.
Honda
S
S
D000631:Aminopyridines; D000983:Antipyrine / Q000031:analogs & derivatives; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D000077553:Edaravone; D004586:Electrophoresis; D006023:Glycoproteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000032:analysis* / Q000737:chemistry; D009844:Oligosaccharides / Q000032:analysis*
D000631:Aminopyridines; D006023:Glycoproteins; D009005:Monosaccharides; D009844:Oligosaccharides; D000077553:Edaravone; D000983:Antipyrine; C032439:alpha-aminopyridine