HIGH-RESOLUTION AND RAPID ANALYSIS OF BRANCHED OLIGOSACCHARIDES BY CAPILLARY ELECTROPHORESIS

The fluorophore 2-aminoacridone has been used to label a number of bra nched oligosaccharides previously released from various glycoproteins. Complex glycans were derivatized via their reducing end with this flu orophore by a Schiff's base mechanism followed by reduction to a secon dary amine using sodium cyanoborohydride. This process of derivatizati on was carried out efficiently and in a nonselective manner over a per iod of 30 min at 90 degrees C. The resulting derivatives were separate d with high resolution by capillary electrophoresis using berate buffe r, containing taurodeoxycholate, as the separation buffer. The method described does not require the removal of sialic acid residues prior t o derivatization, so that treatment of glycans with N-acetyl neuramini dase provided useful and additional structural information. Attempts h ave also been made to relate the electrophoretic mobility of branched oligosaccharides with their molecular volume. (C) 1995 Academic Press, Inc.