HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ANALYSIS OF COMPLEX N-LINKED GLYCANS DERIVATIZED WITH 2-AMINOACRIDONE

2-Aminoacridone (2-AMAC) has been used to derivatize mixtures of N-lin ked oligosaccharides released from alpha(1)-acid glycoprotein and immu noglobulin G. In each case, the HPLC profile obtained for the derivati zed glycans was compared to that obtained after digestion with sialida se and a two-enzyme away system made up of sialidase and fucosidase, p rior to derivatization by 2-AMAC. These studies are rapid and provide a wealth of preliminary information about the degree of sialylation an d core fucosylation in the corresponding parent glycans. Moreover, col lection of glycans from one single injection has provided enough mater ial for molecular weight determination by MALDI-MS analysis. In this s tudy we have also carried out limited MS-MS studies on enriched fracti ons of 2-AMAC-glycans using a nanospray orthogonal quadrupole time-of- night mass spectrometer.