Abstract
A highly sensitive chemical method for detecting α2→8-linked oligo/polysialic acid (oligo/polySia) chains was developed, including (i) periodate oxidation, reduction with sodium borohydride, and subsequent acid hydrolysis, giving rise to C7analogues and intact C9compounds from nonreducing terminal and internal sialic acid residues, respectively; (ii) fluorescent labeling of these C7and C9compounds with 1,2-diamino-4,5-methylenedioxybenzene (DMB); and (iii) quantitation of theseDMB derivatives on fluorometric high-performance liquid chromatography. As little as 1 ng of internal sialic acid residues of oligo/polySia chains, the existence of which indicates the presence of oligo/polySia structure, was detectable by this method. This fluorometric C7/C9analysis was successfully applied to glycoproteins blotted onto a slit of polyvinylidene fluoride membranes and suggested the presence of some novel oligoSia-containing glycoproteins in pig embryonic brains.
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