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Biology Articles » Biochemistry » Carbohydrate Biochemistry » Large-scale approaches for glycobiology » Conventional glycosylation profiling

Conventional glycosylation profiling
- Large-scale approaches for glycobiology

Only recently has methodology advanced sufficiently to obtain complete glycosylation profiles of glycoconjugates such as prions or CD34 (Figure 2). To briefly summarize today's technology, a plethora of mass spectrometry (MS) methods are becoming affordable and user-friendly [17,18], pulsed-amperometric detection methodology is making the separation of carbohydrates by high-pressure liquid chromatography (HPLC) attractive, increasingly sensitive nuclear magnetic resonance (NMR) technology is allowing this powerful technique of structure determination and identification to be applied to glycoconjugates isolated from natural sources, and lectins are finding new uses as detection agents for carbohydrates in chromatography and protein arrays [19-21]. Excellent reviews provide a detailed picture of how different methodologies are coalescing into a powerful set of tools for sophisticated and highly sensitive investigation of glycoconjugates [22,23].

While the isolation and characterization of highly complex glycoproteins are impressive feats, the sobering reality is that only a handful of the thousands of different glycoconjugates in the human body have been analyzed so far, which leaves the enormous carbohydrate diversity of even a single cell unknown in molecular detail. To further complicate matters, glycosylation profiles are not static, but rapidly change as cells differentiate, undergo apoptosis, or become diseased. Today's technologies are inadequate for determining the dynamic glycosylation profile of a cell and fall well short of the ultimate goal of glycomics - the evaluation of an entire organism. To dispel the gloom, however, underlying technologies for innovative, large-scale glycomic techniques are developing rapidly - both by bringing new techniques to carbohydrate analysis and by refining established methods to increase throughput. These two approaches, exemplified by array-based technologies and the automation of mass spectrometry, respectively, are discussed below.


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