C-myc单克隆抗体的制备及鉴定  

Preparation and identification of monoclonal antibodies against C-myc

作　　者：孙慧慧[1] 王旭[2] 王惠国[2] 谭建华[3] 朴敬爱[1] 朴君[1] 马彪[2] 刘庆平[2] 李文哲[2] 

机构地区：[1]辽宁师范大学生命科学学院,大连116029 [2]大连大学生物工程学院免疫研究所,116622 [3]军事医学科学院军事兽医研究所,长春130062

出　　处：《免疫学杂志》2011年第8期688-691,共4页Immunological Journal

基　　金：国家自然科学基金(30972675);大连市科技计划项目(2010J21DW011);国家教育部归国留学人员科研启动资金[教外司留(2008)101]

摘　　要：目的制备与鉴定鼠抗C-myc蛋白单克隆抗体。方法利用C-myc蛋白做免疫原免疫Balb/c小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞进行细胞融合,经间接ELISA法筛选、有限稀释法克隆、单克隆抗体Ig亚型鉴定,获得2株能够稳定分泌抗C-myc特异性抗体的杂交瘤细胞株,再将杂交瘤细胞注射入小鼠腹腔诱使小鼠产生腹水。结果获得2株可稳定分泌抗C-myc抗体的杂交瘤细胞株,命名为SHH-1H11,SHH-2A9,Ig亚型分别为IgG1亚类和IgM。细胞培养上清效价分别为1:10240、1:5 120;小鼠腹水效价分别为1:64 000、1:32 000。结论制备的C-myc单克隆抗体仅与C-myc蛋白反应,与其他蛋白没有交叉反应,表明获得的单克隆抗体的特异性很高。To prepare monoclonal antibodies against for C-myc protein,Balb/c mice were immunized with C-myc protein and the splenocytes of the immunized mice were fused with the SP2/0 myeloma cells by hybridoma technique.Two hybridoma cell lines secreting monoclonal antibodies against C-myc were obtained,which were determined as SHH-1H11,SHH-2A9.The subtypes of antibodies,SHH-1H11 and SHH-2A9 were IgG1 and IgM,respectively.The ELISA titer of antibodies produced by SHH-1H11 and SHH-2A9 were 1：10 240 and 1：5 120 in cell media,and 1：64 000 and 1：32 000 in mice ascites,respectively.

关 键 词：C-MYC 单克隆抗体 制备 鉴定 

分 类 号：R392.11[医药卫生—免疫学]