摘要
本文应用葡聚糖凝胶G-200柱层析和聚丙烯酰胺凝胶板状电泳,将细胞培养上清液及小鼠腹水中的单克隆抗体分离提纯,并对它的理化性质进行了研究.结果表明:①ZMC_9单克隆抗体(小鼠IgM)分子量为871000道尔顿.②纯化单克隆抗体在56℃加热5 min,即失去抗体活性,而未经纯化的单克隆抗体即使加热30min,抗体活性也毫无损失.③纯化单克隆抗体在pH 6～8环境中活性不变,pH超过8.0时活性有所下降,而未经纯化的单克隆抗体在pH 6～10范围内仍表现有活性.④将试剂冻干或加叠氮钠4℃保存,效价至少可维持一年以上.
This study reports the isolation and purification of monoclonal antibody (mouse IgM) from the supernatant of cell-cultured medium and mouse ascites using sephadexG-200 chromatography and polyacrylamide gel electrophoresis. Some of its physico-chemical properties were studied. The results indicated that, (1) The molecules of ZMC_9 monoclonal antibody (mouse IgM) weighed 871 KD approximately. (2) The purified monoclonal antibody completely lost its activity when heated at 56℃ for 5 minutes, but the activity of crude monoclonal antibody was stable even heated for 30 minutes at 56℃. (3) The purified monoclonal antibody kept stable only at a narrow pH range of 6～8. If pH was over 8.0, its activity decreased. But no change was observed in the activity of the crude monoclonal antibody at pH 6～10. (4) The activity of lyophilized or sodium azide preserved antibody stored at 4℃ remained unchange for at least one years.
关键词
ABO血型
单克隆抗体
红细胞
ABO blood-group system
Antibodies monoclonal/diag use
