Fine specificity of anti-Le(X) monoclonal antibody TEC-01

TEC-01 monoclonal antibody recognizes the oligosaccharide structure Galbeta1-->4[Fucalpha1-->3] GlcNAc (Le(X) hapten). To determine its fine specificity, reactivity of TEC-01 antibody with Le(X) glycosphingolipids, isolated from human liver metastasis of adenocarcinoma, and Le(X) neoglycolipid were analysed. Immunostaining of Le(X) glycosphingolipids, fractionated by thin-layer chromatography, showed that TEC-01 reacted with difucosyl and trifucosyl Le(X) glycosphingolipids but not with Le(X) pentasaccharide ceramide. Interestingly, TEC-01 also reacted with Le(X) neoglycolipid prepared by reductive amination from Le(X) pentasaccharide, lacto-N-fucopentaose III, and L-1,2-dipalmitoyl-sn-glycerophosphoethanolamine. The combined data suggest that TEC-01 recognizes Le(X) hapten in glycolipids with longer oligosaccharide chain moiety.