GENERATION OF ONE SET OF MONOCLONAL-ANTIBODIES SPECIFIC FOR B-PATHWAY GANGLIO-SERIES GANGLIOSIDES
GENERATION OF ONE SET OF MONOCLONAL-ANTIBODIES SPECIFIC FOR B-PATHWAY GANGLIO-SERIES GANGLIOSIDES
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DOI：
10.1016/0005-2760(92)90110-h
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发表时间：
1992-01-24
期刊：
影响因子：
--
通讯作者：
TAI, T
中科院分区：
文献类型：
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作者：
OZAWA, H;KOTANI, M;TAI, T
我们通过用吸附在明尼苏达沙门氏菌突变株R595上的纯化神经节苷脂免疫C3H/HeN小鼠，建立了6种对b -途径神经节系列神经节苷脂具有特异性的鼠单克隆抗体（MAb）。通过酶联免疫吸附测定和薄层色谱免疫染色确定了这些单克隆抗体的结合特异性。这6种单克隆抗体，分别命名为GGB19、GMR2、GMR7、GGR12、GMR5和GGR13，分别与用作免疫原的神经节苷脂GD3、O - Ac - GD3、GD2、GD1b、GT1b和GQ1b强烈反应。除GGB19外，所有这些单克隆抗体都显示出高度受限的结合特异性，仅与免疫所用的神经节苷脂反应。不识别其他各种天然神经节苷脂或中性糖脂。另一方面，单克隆抗体GGB19表现出更广泛的特异性，与O - Ac - GD3、GQ1b和GT1a有微弱的交叉反应，但不与其他神经节苷脂或中性糖脂反应。利用这些单克隆抗体，我们测定了这些神经节苷脂，特别是GD1b、GT1b和GQ1b在小鼠、大鼠和人白血病细胞上的表达。GD1b在大鼠白血病细胞上表达，但在所检测的小鼠和人白血病细胞上不表达。在这些细胞系中未检测到GT1b和GQ1b。
We established six murine monoclonal antibodies (MAbs) specific for b-pathway ganglio-series gangliosides by immunizing C3H/HeN mice with these purified gangliosides adsorbed to Salmonella minnesota mutant R595. The binding specificities of these MAbs were determined by an enzyme-linked immunosorbent assay and immunostaining on thin-layer chromatogram. These six MAbs, designated GGB19, GMR2, GMR7, GGR12, GMR5, and GGR13 reacted strongly with the gangliosides GD3, O-Ac-GD3, GD2, GD1b, GT1b, and GQ1b, respectively, that were used as immunogens. All these MAbs except GGB19 showed highly restricted binding specificities, reacting only with the immunizing ganglioside. None of other various authentic gangliosides or neutral glycolipids were recognized. On the other hand, MAb GGB19 exhibited a broader specificity, cross-reacting weakly with O-Ac-GD3, GQ1b, and GT1a, but not with other gangliosides or neutral glycolipids. Using these MAbs, we determined the expression of these gangliosides, especially GD1b, GT1b, and GQ1b on mouse, rat, and human leukemia cells. GD1b was expressed on rat leukemia cells, but not on mouse and human leukemia cells tested. Neither GT1b nor GQ1b was detected in these cell lines.