Optimising the separation of derivatised oligosaccharides and N-glycopeptides of human serum immunoglobulin G using hydrophilic interaction chromatography
The separation of derivatised oligosaccharides (glycans) and N-glycopeptides of human serum immunoglobulin G (IgG) can be readily effected via hydrophilic-interaction chromatography using Zic-Hilic, an extremely versatile zwitterionic stationary phase which includes a sulfobetaine group, according to professor Hiroaki Nakagawa of the division of biological sciences, graduate school of science, Hokkaido University, Sapporo, Japan.
This separation is one of the more challenging issues in liquid chromatography when reverse phase (RP) or normal phase (NP) chromatography is used.
There are several reasons for this, including the structural diversity of the compounds (composition, sequence, anomeric character, linkage position, and branching pattern) and the existence of many isomers.
The Zic-Hilic column that was used has a very high capability for the structural recognition of isomeric N-glycans as well as high selectivity for glycopeptides.
IgG glycopeptides consisting of isomeric N-glycans and the same peptide sequences can be easily separated on a Zic-Hilic column and the column is highly promising for a simple analysis of N-glycans and N-glycopeptide samples.
This method is capable to separate N-glycopeptide isomers and is particularly useful for a LC/MS based direct analysis of peptide amino-acid sequence and isomeric N-glycan structures of N glycopeptides.
Zic-Hilic hydrophilic interaction sorbents provide separations that are rapid, sensitive, provide superb resolution and are readily scalable for preparative work.
Typical applications include the separations of polar drugs, glycosylated peptides, purines and pyrimidines, nucleotides, peptides, tryptic digests, acrylamides, quaternary amines, organic acids, morphine and flavanoids from complex samples.
