Glycoprotein-Lectin-Immunosorbent Assay (GLIA)
Much reported work in recent years points to the essential physiological and pathobiochemical importance of the regulated expression of the variable glycosylation of proteins and lipids. (Schachter 1984; Martinez and Barsigian 1987). The structural analysis of the glycans of single purified glycoproteins requires, however, a range of extraordinarily complicated and costly methods. For this reason, the investigation of changes in protein glycosylation in body fluids or tissue homogenates is often not performed on individually purified glycoproteins, but on glycoprotein mixtures. An important contribution to such studies has been made by lectin-aifinity chromatography (Osawa and Tsuji 1987), but this technique presents certain difficulties when applied to the analysis of very small samples, or large sample numbers. As an alternative approach — usually with limited success — attempts have been made to relate the biochemical or pathobiochemical regulatory role of glycoproteins to the total content of distinct sugar residues, e.g., of Neu5Ac or fucose in glycoprotein mixtures. With the technique described here it is possible to selectively analyze the glycans of defined glycoproteins without prior purification. The method displays high sensivity, and requires only small sample quantities; it is quick to perform, and the operational costs are favorable (Köttgen et al. 1988, 1989). In addition, the necessary apparatus is available in practically all modern biochemical laboratories.
KeywordsSugar DMSO Citrate Fractionation Polypeptide
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