title: Monoclonal antibody R24 distinguishes between different N-acetyl- and N-glycolylneuraminic acid derivatives of ganglioside GD3.
abstract: Monoclonal antibody (MAb) R24 was previously shown to be directed toward ganglioside GD3 [Pukel, C. S., Lloyd, K. O., Travassos, L. R., Dippold, W. G., Oettgen, H. F., and Old, L. J. (1982) J. Exp. Med. 155, 1133-1147]. The structural specificity of the MAb has now been further characterized based on binding to structurally related glycolipids, including four GD3 derivatives with different N-acetylneuraminic acid (NeuAc) and N-glycolylneuraminic acid (NeuGc) substituents. Three assay systems (enzyme immunostaining on thin-layer chromatography, enzyme-linked immunosorbent assay, and immune adherence inhibition assay) were used. MAb R24 was found to react with (NeuAc-NeuAc-)GD3 and (NeuAc-NeuGc-)GD3 but not with (NeuGc-NeuAc-)GD3 or (NeuGc-NeuGc-)GD3. These results clearly indicate that the outer sialic acid (Sia) moiety of GD3 is crucial and must be a NeuAc residue, while the inner sialic acid is less involved in binding to the MAb and can be either NeuAc or NeuGc. The MAb was also found to cross-react weakly with two gangliosides, GT1a and GQ1b, but none of other gangliosides nor neutral glycolipids tested reacted. These findings suggest that the epitope detected by MAb R24 is the trisaccharide structure NeuAc alpha 2----8Sia alpha 2----3Gal-, which must be in a terminal position.
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D000937:Antigen-Antibody Reactions; D002417:Cattle; D002855:Chromatography, Thin Layer; D000939:Epitopes / Q000276:immunology; D005732:Gangliosides / Q000276:immunology*; D006017:Glycolipids / Q000276:immunology; D006801:Humans; D007957:Leukocyte Adherence Inhibition Test; D051379:Mice; D019158:N-Acetylneuraminic Acid; D012794:Sialic Acids / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D005732:Gangliosides; D006017:Glycolipids; D012794:Sialic Acids; C026226:ganglioside, GD3; D019158:N-Acetylneuraminic Acid