Glycosphingolipids Covalently Linked to Agarose Gel or Glass Beads

Oxidative ozonolysis of the olefinic bond of the sphingosine moiety of either globoside (βGalNAc1→3αGal1→4βGal1→ 4βGlc→ceramide), or of the methyl ester of hematoside (αN-glycolylneuraminyl2→3βGal1→4βGlc→ceramide), and coupling of the carboxyl-bearing product to aminoethylagarose or to amino group-bearing glass beads in the presence of a carbodiimide resulted in globoside or hematoside covalently linked to agarose or glass beads. These compounds were used for purification of anti-glycosphingolipid antibodies from serum of immunized rabbits. The antibodies bound to the substrate were released by 1 m sodium iodide and their immunological properties were studied. Anti-globoside is directed toward the terminal β-(N-acetyl)galactosaminosyl-(1→3)α-galactopyranosyl structure, while anti-hematoside is directed predominantly toward the sialosyl residue of hematoside.