鼠抗早孕因子单克隆抗体的制备及鉴定  

Preparation and iIdentification of murine monoclonal antibody against EPF

作　　者：练玉银[1] 吴晓亮[1] 王家骥[1] 温志锋[1] 聂湘辉[1] 

机构地区：[1]广州医学院公共卫生学院,广东广州510182

出　　处：《中国热带医学》2012年第5期532-534,共3页China Tropical Medicine

基　　金：广东省科技计划重点项目(No.2008A030201022)

摘　　要：目的制备鼠抗早孕因子(EPF)单克隆抗体,纯化并进行鉴定。方法用重组EPF免疫BALB/c小鼠,采用杂交瘤技术获得抗EPF杂交瘤细胞株,注入BALB/c小鼠腹腔制备腹水型单抗,Protein-G亲和层析纯化后,SDS-PAGE电泳鉴定其纯度,用间接ELISA法测抗体效价,ELISA相加试验测定抗原表位,用Western blotting鉴定其特异性,并进行类及亚类、细胞染色体核型分析。结果筛选出2株具有较高活性和良好稳定性的分泌抗EPF单克隆抗体的杂交瘤细胞株,其腹水的抗体效价均高于2×10-5,2株单抗均为IgG类IgG1亚类,轻链类型均为κ,细胞核型分析符合杂交瘤细胞特征,特异性良好,2株单抗有不同的抗原表位。SDS-PAGE电泳显示纯化后抗体有较高的纯度,Western blotting表明其与EPF抗原有特异结合性。结论成功制备2株具有较高活性的鼠抗EPF单克隆抗体,为建立检测EPF方法奠定了良好的基础。Objective To prepare and identify the murine monoclonal antibody（McAb）against early pregnancy factor（EPF）.Methods BALB/c mice were immunized with recombinant EPF,based on which the McAb was prepared by hybridoma technique,then purified and determined for titer by indirect ELISA,for antigenic epitopes by additive ELISA,for specificity by Western blotting,and analyzed for class,subclass and chromosome karyotypes.Results Two cell strains secreting McAb against EPF were screened.The McAbs showed high activity.Both two McAbs were IgG1 with κ chain,and their titers were higher than 2×10^-5 in ascites.The chromosome karyotypes of the 2 cell strains were consistent with those of hybridoma cells.The antigenic epitopes of two McAbs were different.McAbs have high purity and specificity detected by SDS-PAGE and Western blotting respectively.Conclusion The murine McAbs against EPF with high activity were successfully prepared.

关 键 词：早孕因子 单克隆抗体 鉴定 

分 类 号：R714.15[医药卫生—妇产科学]