Quantitative recovery of Man9GlcNAc2Asn derivatives from concanavalin A

Some larger high-mannose-type oligosaccharides bind very tightly to concanavalin A and are difficult to elute. We present conditions that permit the complete elution of compounds containing high-mannose type oligosaccharides from a popular concanavalin A-Sepharose that is commercially available. Europium-labeled Man9GlcNAc2Asn (N. Kawasaki and Y.C. Lee, Anal. Biochem., 250 (1997) 260-262), soybean agglutinin, and Eu(III)-labeled soybean agglutinin bound to concanavalin A-Sepharose were completely eluted with 1 M methyl alpha-D-mannopyranoside by allowing the column to stand in elution buffer, permitting the oligosaccharide or glycoprotein to slowly dissociate from the column.