CE-MS Analysis of High Mannose Glycans and Isomers
Date Issued
2011
Date
2011
Author(s)
Lu, En-Tzu
Abstract
Separation of glycans is necessary before analyzed by MS for their structural complexity. A comprehensive study of glycan structure could be improved by analyzed though CE-MS. The glycans of RNase B, categorized as high-mannose type oligosaccharides, were chosen as the model compounds. The glycans released from RNase B were labeled with p-aminobenzoic acid (ABA) and 8-aminopyrene-1,3,6-trisulfonic acid (APTS) either by reductive or non-reductive amination. Two kinds of electrophoresis system, including CZE using borate buffer and CGE using acetate buffer with polyethylene oxide (PEO), were selected to analyze the glycans. In borate buffer system, normal polarity was employed under high pH environment. In the acetate buffer with polyethylene oxide (PEO), the reverse polarity was employed under low pH environment. According to the premier results using CE-UV, in both CZE and CGE system, the better separation can be obtained for glycans labeled with APTS by reductive amination can be obtained with better separation results comparing to glycans labeled with APTS by non-reductive amination and ABA labeling either by reductive or non-reductive amination. To avoid ion suppression by borate ions and PEO as CE was coupled to MS, a double junction interface was applied. By employing the double junction interface, which was composed of a liquid junction, a connecting column and a low-flow interface, borate and PEO were restricted in the liquid junction, while the analytes were detected successfully and the isomers structure of M7G2 were analyzed by MS.
Subjects
CE-MS
high-mannose glycans
APTS
borate buffer
PEO
double junction interface
isomers
Type
thesis
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