원문정보
초록
영어
Mass spectrometry (MS) is the single most powerful enabling tool for its sensitivity and precision in glycomic mapping. It is well suited for systematic exploratory and discovery investigations since it alone is not pre-defined by availability of biological probes raised against already known molecular entities. In practice though, it is limited by the need to resolve a great depth of isomeric variation at a wide range of abundance, particularly those of larger size extended by polylactosaminoglycans, and those carrying multiple sialic acids and sulfates. Glycomic mapping, as most often employed to define glycosylation differences between genetically manipulated cell types or onco-developmental stages, therefore entails multi-staged MS analyses at increasing levels of sophistications and technical demands, in conjunction with chemo-enzymatic derivatization and chromatographic separation. A most effective analysis that is of true glycobiology significance is one that aims to address specifically if a particular glycotope of interest is expressed at what level on which glycan and protein carriers. Thus following a first-screen MALDI MS mapping and selected MALDI-MS/MS glycan sequencing to define the overall profile, further rounds of nanoESI-based analysis aiming at both comprehensiveness and selectivity in glycomic coverage are implemented. Along this line, our concerted workflows in glycomics and proteomics, as employed in recent case studies, will be presented to drive home this conceptual framework.