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REVIEW: High-Performance Anion-Exchange Chromatography with Pulsed Amperometric Detection for Carbohydrate Analysis of Glycoproteins

J. S. Rohrer*, L. Basumallick, and D. Hurum

Thermo Fisher Scientific, 1214 Oakmead Parkway, Sunnyvale, CA, USA; fax: 408-730-9403; E-mail: Jeff.Rohrer@thermofisher.com

* To whom correspondence should be addressed.

Received January 28, 2013
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).
KEY WORDS: monosaccharide, sialic acid, oligosaccharide, glycoprotein, carbohydrate, mannose-6-phosphate, HPAE-PAD

DOI: 10.1134/S000629791307002X