N-Linked Oligosaccharide Profiling by HPAEC-PAD
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High-pH anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) is a very powerful tool for the profiling of N-linked oligosaccharides (1,2), only being limited by the concentrated sodium hydroxide and sodium acetate required to achieve the separation and sensitive detection. Oligosaccharides are separated on the basis of charge (i.e., the number of sialic acid residues) and the linkage isomers present. Neutral oligosaccharide alditols are most weakly retained, retention increasing with increasing sialylation, and NeuGc- and sulfate-bearing oligosaccharides being most strongly retained. Fucosylation results in a shorter retention. If the oligosaccharides can be effectively desalted after HPAEC-PAD, this remains the method of choice for oligosaccharide purification, and is a very powerful analytical tool.
KeywordsSodium Acetate Sialic Acid Residue Pulse Amperometric Detection Powerful Analytical Tool Dynamic Mixer