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A chromatographic and mass spectrometric strategy for the analysis of oligosaccharides: determination of the glycan structures in porcine thyroglobulin
Author(s) -
Charlwood Joanne,
Birrell Helen,
Organ Andrew,
Camilleri Patrick
Publication year - 1999
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/(sici)1097-0231(19990430)13:8<716::aid-rcm547>3.0.co;2-c
Subject(s) - chemistry , chromatography , glycan , mass spectrometry , capillary electrophoresis , sialidase , thyroglobulin , high performance liquid chromatography , time of flight mass spectrometry , matrix assisted laser desorption/ionization , desorption , enzyme , biochemistry , glycoprotein , organic chemistry , adsorption , neuraminidase , ionization , ion , antibody , immunology , biology
Oligosaccharides released from porcine thyroglobulin were first derivatised with 2‐aminoacridone (2‐AMAC) and analysed by capillary electrophoresis to determine the complexity of this glycan pool. The same glycan mixture was then subjected to either a sialidase digest or a sialidase and fucosidase digest prior to derivatisation with 2‐AMAC and analysis by normal phase high pressure liquid chromatography (HPLC). Comparison of the three chromatographic profiles with known standards allowed an initial identification of the glycan structures. The 2‐AMAC derivatised glycans were then collected from HPLC for matrix‐assisted laser desorption/ionisation time‐of‐flight (MALDI‐TOF) analysis and the molecular weights of predicted structures were confirmed. This study demonstrates that a two enzyme array and subsequent MALDI‐TOF analysis can be used successfully to assign the major glycans present in a complex mixture. Copyright © 1999 John Wiley & Sons, Ltd.