两株鼠抗人PD-1单克隆抗体的制备及其生物学特性鉴定  

Preparation of two mouse anti-human PD-1 monoclonal antibodies and identification of their biological characteristics

作　　者：明志君[1,2] 吴海竞[2] 张光波[1] 胡玉敏[1] 邱玉华[1] 张学光[1] 

机构地区：[1]苏州大学医学生物技术研究所江苏省临床免疫研究室,江苏苏州215123 [2]苏州大学医学部药学院,江苏苏州215123

出　　处：《细胞与分子免疫学杂志》2009年第6期501-503,506,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30330540);苏州大学医学发展基金(EE132030);苏州大学在职博士学位基金资助(13132743)

摘　　要：目的:制备鼠抗人PD-1单克隆抗体(mAb)及鉴定其生物学特性。方法:以基因转染细胞PD-1/L929为免疫原,免疫BALB/c小鼠;采用淋巴细胞杂交瘤技术进行细胞融合,经选择性克隆化培养及流式细胞术(FCM)分析,筛选分泌鼠抗人PD-1分子mAb的杂交瘤细胞株;采用Ig亚型快速定性试纸法、染色体核型分析、Western blot、竞争结合抑制试验及肿瘤细胞株测定等方法对mAb进行生物学特性鉴定。结果:获得了2株持续稳定分泌鼠抗人PD-1mAb的杂交瘤细胞株,命名为1F2和5F10。对其生物学特性的鉴定结果表明,均为条带相对分子质量在(Mr)55000左右的免疫球蛋白,具有不同的PD-1结合位点;1F2mAb可识别SKHep-1和7721细胞株表面的PD-1分子,而5F10可识别Raji细胞株的PD-1分子。结论:成功获得了2株鼠抗人PD-1杂交瘤及其分泌的mAb。AIM：To prepare mouse anti-human PD-1 monoclonal antibodies （mAbs） and identify their biological characteristics. METHODS：The BALB/c mice were immunized with the transfected cell line PD-1/L929. The cells were fused with Sp2/0 using monoclonal antibody techniques and the positive clones were screened by FCS with PD-1/L929. The secreted anti-PD-1 mAbs were identified through rapid isotyping analysis,karyotype analysis,Western blot,competitive inhibition test,indirect immunofluorescence assay,and tumor cell lines detection. RESULTS：Two mouse anti-human PD-1 hybridomas were obtained and their secreted mAbs were named （1F2 and 5F10）. Their biological characteristics suggested that they could recognize a protein with approximate molecular weight 55 000 on PD-1/L929 cell lines and different epitopes. 1F2 could recognize PD-1 molecules expressed on SKHep-1 and 7721 while 5F10 could recognize Raji cells. CONCLUSION：Two mouse anti-human PD-1 hybridoma cell lines and their secreted monoclonal antibodies have been successfully obtained and identified.

关 键 词：PD-1 杂交瘤 单克隆抗体 

分 类 号：R392.12[医药卫生—免疫学]