Analysis of high-mannose-type oligosaccharides by microliquid chromatography-mass spectrometry and capillary electrophoresis

被引：22
作者：
Koller, A [1 ]
Khandurina, J [1 ]
Li, JC [1 ]
Kreps, J [1 ]
Schieltz, D [1 ]
Guttman, A [1 ]
机构：
[1] Diversa Corp, San Diego, CA 92121 USA
来源：
ELECTROPHORESIS | 2004年 / 25卷 / 13期
关键词：
capillary electrophoresis; glycopeptides; microbore liquid chromatography-mass spectrometry; oligosaccharides;
D O I：
10.1002/elps.200305837
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
We report on microbore liquid chromatography (muLC) 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 muLC/electrospray ionization/mass spectrometry (ESI-MS) and muC/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 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 muLC/ESI-MS and MS/MS data with respect to the possible number of positional isomers.
引用
收藏
页码：2003 / 2009
页数：7
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