title: High-performance anion-exchange chromatography with pulsed amperometric detection for carbohydrate analysis of glycoproteins.
abstract: High-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD) is an established technique for the carbohydrate analysis of glycoproteins. HPAE-PAD is routinely used for determinations of monosaccharide, sialic acid, mannose-6-phosphate (M-6-P), and oligosaccharide contents of a glycoprotein. This is true for both the initial investigation of a glycoprotein and routine assays of recombinant therapeutic glycoproteins. This contribution reviews the fundamentals of HPAE-PAD, recent technological improvements, and advances in the last ten years in its application to carbohydrate analysis of glycoproteins. The application areas reviewed include monosaccharide determinations, sialic acid determinations, M-6-P determinations, sugar alcohol determinations, analysis of polysialic acids, neutral and charged oligosaccharide analysis, following glycosidase and glycosyltransferase reactions, and coupling HPAE-PAD to mass spectrometry (MS).
authors.affiliation: Thermo Fisher Scientific, CA, USA. Jeff.Rohrer@thermofisher.com
authors.forename: J S
authors.initials: JS
authors.lastname: Rohrer
authors.forename: L
authors.initials: L
authors.lastname: Basumallick
authors.forename: D
authors.initials: D
authors.lastname: Hurum
mesh_terms: D002241:Carbohydrates / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D055664:Electrochemical Techniques / Q000379:methods*; D006023:Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D008360:Mannosephosphates / Q000032:analysis; D013058:Mass Spectrometry; D009005:Monosaccharides / Q000032:analysis; D009844:Oligosaccharides / Q000032:analysis; D012794:Sialic Acids / Q000032:analysis
chemical_list: D002241:Carbohydrates; D006023:Glycoproteins; D008360:Mannosephosphates; D009005:Monosaccharides; D009844:Oligosaccharides; D012794:Sialic Acids; C027693:mannose-6-phosphate