Abstract
Spleen cells from NZB mouse immunized with a membrane fraction of rabbit thymus tissue were fused with BALB/c 6-thioguanine-resistant myeloma cells, P3-X63-Ag8.653. One hybridoma clone (Y-2-HD-1) produced IgM immunoglobulin that bound to an N-glycolylneuraminic acid-containing G M2 ganglioside, G M2(NeuGc), which is known to be a Hanganutziu-Deicher antigen. The specificity of the Y-2-HD-1 monoclonal antibody was examined, using authentic glycosphingolipids structurally related to G M2(NeuGc), by means of an enzyme-linked immunosorbent assay and thin-layer chromatography/enzyme immunostaining, respectively. The monoclonal antibody was found to be highly specific to G M2(NeuGc) and the epitope was a non-reducing terminal GalNAcβ1–4[NeuGcα2–3]Gal structure. This monoclonal antibody (Y-2-HD-1) bound to native mouse erythrocytes, in which G M2(NeuGc) is a major ganglioside. These results indicate that G M2(NeuGc) is located on the surface of mouse erythrocytes.
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