抗人IgM重链单克隆抗体的特性分析

本文报道取正常人混合血清纯化 IgM,经断链后获得(Fo)_5μ片段作为免疫原免疫动物,应用杂交瘤技术获得16株分泌抗人IgM McAb的细胞株。用琼脂双扩散、免疫电泳、ELISA等方法证实所分泌的 McAb对人 IgM 呈高特异性。杂交瘤细胞诱生腹水的抗体效价在双扩散试验中可达1:256,在ELISA 中可达10~(-7)。用被动血凝抑制试验及ELISA试验比较了其中4种McAb(M_5、M_(12)、M_(23)、M_(25))的抗原结合力大小,表明M_(12)有较高亲和力。用ELISA固相竞争试验分析,证明该4种McAb针对同一抗原的不同决定簇。该 16种McAb除 M_(22)为 IgA外,余均属 IgG1亚类。

THE CHARACTERIZATION OF MONOCLONAL ANTIBODIES AGAINST HUMAN IgM HEAVY CHAIN

We have succeeded in establishment of sixteen hybridoma cell lines secreting anti-human IgM McAb, and undertook a detailed characterization of these monoclonal antibodies.The specificity of the McAbs against IgM was confirmed by immunoelectrophoresis double diffusion test,PHA and ELISA. No reactivity was detected with human IgA, IgG, x and λ light chains. Mouse ascitic fluids produced by these hybridomas showed different reactivity in different assays. All of these fluids had antibody tilers against human IgM of 10-4-10-5 by PHA,though only six of them had antibody titers of 10-7-10-7 by ELISA. All of sixteen monoclonal antibodies belonged to the mouse IgGl subclass but M22 which had IgA property.