Sanai Y, Yamasaki M, Nagai Y
Department of Biochemistry, Faculty of Medicine, University of Tokyo, Japan.
Biochim Biophys Acta. 1988 Feb 19;958(3):368-74. doi: 10.1016/0005-2760(88)90222-6.
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 GM2 ganglioside, GM2(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 GM2(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 GM2(NeuGc) and the epitope was a non-reducing terminal GalNAc beta 1-4[NeuGc alpha 2-3]Gal structure. This monoclonal antibody (Y-2-HD-1) bound to native mouse erythrocytes, in which GM2(NeuGc) is a major ganglioside. These results indicate that GM2(NeuGc) is located on the surface of mouse erythrocytes.
用兔胸腺组织膜部分免疫的NZB小鼠的脾细胞与BALB/c 6-硫代鸟嘌呤抗性骨髓瘤细胞P3-X63-Ag8.653融合。一个杂交瘤克隆（Y-2-HD-1）产生与含N-羟乙酰神经氨酸的GM2神经节苷脂GM2(NeuGc)结合的IgM免疫球蛋白，GM2(NeuGc)已知是汉努齐乌-戴歇尔抗原。分别通过酶联免疫吸附测定和薄层色谱/酶免疫染色，使用与GM2(NeuGc)结构相关的真实糖鞘脂来检测Y-2-HD-1单克隆抗体的特异性。发现该单克隆抗体对GM2(NeuGc)具有高度特异性，表位是一个非还原末端GalNAcβ1-4[NeuGcα2-3]Gal结构。这种单克隆抗体（Y-2-HD-1）与天然小鼠红细胞结合，其中GM2(NeuGc)是主要的神经节苷脂。这些结果表明GM2(NeuGc)位于小鼠红细胞表面。
