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Analysis of high‐mannose‐type oligosaccharides by microliquid chromatography‐mass spectrometry and capillary electrophoresis
Author(s) -
Koller Antonius,
Khandurina Julia,
Li Jincai,
Kreps Joel,
Schieltz David,
Guttman András
Publication year - 2004
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200305837
Subject(s) - chromatography , capillary electrophoresis , mass spectrometry , mannose , chemistry , capillary electrophoresis–mass spectrometry , electrospray ionization , biochemistry
We report on microbore liquid chromatography (μLC) and capillary electrophoresis (CE) separation of glycopeptides and high‐mannose‐type oligosaccharides, digested from recombinant phospholipase C, expressed in Pichia pastoris. The glycopeptides were subject to μLC/electrospray ionization/mass spectrometry (ESI‐MS) and μLC/ESI‐tandem MS (MS/MS) analysis that revealed high‐mannose structure size variation between Man 7 GlcNAc 2 and Man 14 GlcNAc 2 . Then, high‐performance CE was applied to identify possible positional isomers of the high‐mannose structures. For the CE experiments, the oligosaccharides were released from the glycoproteins by peptide‐ N ‐glycosidase F and labeled with 1‐aminopyrene‐3,6,8‐trisulfonic acid (APTS). Excellent separation of the possible positional isomers was attained, suggesting one for Man 9 GlcNAc 2 , two for Man 10 GlcNAc 2 , three for Man 11 GlcNAc 2 , Man 12 GlcNAc 2 , and Man 13 GlcNAc 2 , and two for Man 14 GlcNAc 2 . The CE results provided complementary information to the μLC/ESI‐MS and MS/MS data with respect to the possible number of positional isomers.