Tai T, Kawashima I, Tada N, Dairiki K
Department of Oncology, Tokyo Metropolitan Institute of Medical Science.
J Biochem. 1988 Apr;103(4):682-7. doi: 10.1093/oxfordjournals.jbchem.a122329.
The fine structural specificities of six monoclonal antibodies (MAbs) to ganglioside GD2, GalNAc beta 1----4(NeuAc alpha 2----8NeuAc alpha 2----3)Gal beta 1----4Glc-Cer, were studied. The binding specificities of these MAbs were found to differ from each other by virtue of their binding to structurally related authentic standard glycolipids as revealed by three different assay systems, including enzyme immunostaining on thin-layer chromatography, enzyme-linked immunosorbent assay, and immune adherence inhibition assay. The MAbs examined could be divided into three binding types. MAbs A1-201, A1-410, and A1-425 bound specifically to ganglioside GD2 and none of the other gangliosides tested. Two other MAbs (A1-245 and A1-267) reacted not only with GD2, but also with several other gangliosides having the sequence NeuAc alpha 2----8NeuAc alpha 2----3Gal (GD3, GD1b, GT1a, GT1b, and GQ1b). The reactivities with these gangliosides varied to some degree. In addition, these MAbs were found to react with both GD3(NeuAc-NeuAc) and GD3(NeuGc-NeuAc), but not with GD3(NeuAc-NeuGc) or GD3(NeuGc-NeuGc). The last MAb (A1-287) also reacted with several other gangliosides but with lower avidity than A1-245 and A1-267. These findings suggest that each MAb to ganglioside GD2 may have an individual binding specificity and avidity. These MAbs represent potentially useful reagents for analyzing the function of GD2 on cell surface membranes, and provide a system for precisely studying the interactions between an anti-ganglioside antibody and the binding epitope of the antigenic determinant.
研究了六种针对神经节苷脂GD2（GalNAcβ1----4(NeuAcα2----8NeuAcα2----3)Galβ1----4Glc-Cer）的单克隆抗体（MAb）的精细结构特异性。通过三种不同的检测系统，包括薄层色谱上的酶免疫染色、酶联免疫吸附测定和免疫黏附抑制测定，发现这些单克隆抗体的结合特异性因其与结构相关的真实标准糖脂的结合而彼此不同。所检测的单克隆抗体可分为三种结合类型。单克隆抗体A1-201、A1-410和A1-425特异性结合神经节苷脂GD2，而不与所检测的其他神经节苷脂结合。另外两种单克隆抗体（A1-245和A1-267）不仅与GD2反应，还与其他几种具有NeuAcα2----8NeuAcα2----3Gal序列的神经节苷脂（GD3、GD1b、GT1a、GT1b和GQ1b）反应。与这些神经节苷脂的反应程度有所不同。此外，发现这些单克隆抗体与GD3(NeuAc-NeuAc)和GD3(NeuGc-NeuAc)均反应，但不与GD3(NeuAc-NeuGc)或GD3(NeuGc-NeuGc)反应。最后一种单克隆抗体（A1-287）也与其他几种神经节苷脂反应，但亲和力低于A1-245和A1-267。这些发现表明，每种针对神经节苷脂GD2的单克隆抗体可能具有独特的结合特异性和亲和力。这些单克隆抗体是分析GD2在细胞表面膜上功能的潜在有用试剂，并为精确研究抗神经节苷脂抗体与抗原决定簇结合表位之间的相互作用提供了一个系统。
