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  • Title: Profiling of N- and O-glycopeptides of erythropoietin by capillary zwitterionic type of hydrophilic interaction chromatography/electrospray ionization mass spectrometry.
    Author: Takegawa Y, Ito H, Keira T, Deguchi K, Nakagawa H, Nishimura S.
    Journal: J Sep Sci; 2008 May; 31(9):1585-93. PubMed ID: 18435510.
    Abstract:
    Capillary zwitterionic-type hydrophilic interaction chromatography (ZIC-HILIC)/ESI-MS has been applied to the Glu-C digest of recombinant human erythropoietin (rhEPO) expressed in Chinese hamster ovary (CHO) cells. N-Glycopeptides (105) and O-glycopeptides (8) were detected in a single run of the capillary ZIC-HILIC/ESI-MS analysis. Among them, N-acetyl-neuraminic acids (Neu5Ac) of N- and O-glycans were partially acetylated and some were replaced with N-glycoyl-neuraminic acid (Neu5Gc). Their retentions in the ZIC-HILIC separation can be explained to some extent with the degree of acetylation and N-glycoylation, both of which influence the hydrophilicity/hydrophobicity of the N- and O-glycan moieties of glycopeptides.
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