Premium
Structural determination of N ‐linked glycans by matrix‐assisted laser desorption/ionization and electrospray ionization mass spectrometry
Author(s) -
Harvey David J.
Publication year - 2005
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200401248
Subject(s) - chemistry , mass spectrometry , electrospray ionization , sample preparation in mass spectrometry , chromatography , protein mass spectrometry , electrospray , fragmentation (computing) , capillary electrophoresis–mass spectrometry , desorption electrospray ionization , extractive electrospray ionization , matrix assisted laser desorption/ionization , top down proteomics , glycan , analytical chemistry (journal) , ionization , ion , desorption , chemical ionization , organic chemistry , biochemistry , adsorption , computer science , glycoprotein , operating system
This paper reviews methods for the analysis of N ‐linked glycans by mass spectrometry with emphasis on studies conducted at the Oxford Glycobiology Institute. Topics covered are the release of glycans from sodium dodecyl sulphate‐polyacrylamide gel electrophoresis gels, their purification for analysis by mass spectrometry, methods based on matrix‐assisted laser desorption/ionization (MALDI) and electrospray ionization for producing fragment ions, and details of their fragmentation. MALDI mass spectrometry provided a rapid method for profiling neutral N ‐linked glycans as their [M + Na] + ions which could be fragmented by collision‐induced decomposition to give spectra containing both glycosidic and cross‐ring fragments. Electrospray ionization mass spectrometry was more versatile in that it was relatively easy to change the type of ion that was formed and, furthermore, unlike MALDI, electrospray did not cause extensive loss of sialic acids from sialylated glycans. Negative ions formed by addition of anions such as chloride and, particularly, nitrate, to the electrospray solvent were stable and enabled singly charged ions to be obtained from larger glycans than was possible in positive ion mode. Fragmentation of negative ions followed specific pathways that defined structural details of the glycans that were difficult to obtain by classical methods such as exoglycosidase digestion.