High-performance anion-exchange chromatography of asparagine-linked oligosaccharides.
Oligosaccharides released enzymatically by N-glycanase from fetuin, alpha-acid glycoprotein, human chorionic gonadotropin, platelet-derived growth factor, and kallikrein were chromatographed on a polymeric pellicular anion-exchange column at pH values of 5 and 13. Separations occurred into groups of peaks containing the same number of sialic acids with an additional separation dependent upon the nature of the antennary structure present. High pH conditions were required for the optimum separation of fetuin oligosaccharides, while low pH conditions significantly improved resolution of oligosaccharides obtained from the other glycoproteins. The analytical separation of oligosaccharides under conditions of low pH has important implications in the development of chromatographic mapping and identification techniques for N-linked oligosaccharides present on recombinant proteins.
D000818:Animals; D001216:Asparagine / Q000737:chemistry*; D016466:CHO Cells; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006224:Cricetinae; D006023:Glycoproteins / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007610:Kallikreins / Q000737:chemistry; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry; D009961:Orosomucoid / Q000737:chemistry; D010982:Platelet-Derived Growth Factor / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D000509:alpha-Fetoproteins / Q000737:chemistry
D006023:Glycoproteins; D009844:Oligosaccharides; D009961:Orosomucoid; D010982:Platelet-Derived Growth Factor; D011994:Recombinant Proteins; D000509:alpha-Fetoproteins; D001216:Asparagine; D007610:Kallikreins