抗GST单克隆抗体的制备及应用  

Production and application of monoclonal antibodies to GST.

作　　者：吴建祥[1] 陈天佳[2] 周雪平[1] 

机构地区：[1]浙江大学生物技术研究所,浙江杭州310029 [2]浙江大学校医院,浙江杭州310029

出　　处：《浙江大学学报（农业与生命科学版）》2006年第4期383-386,共4页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：杭州市科技发展计划资助项目(2003132B22);国家自然科学基金重点资助项目(30530520)

摘　　要：用纯化的重组GST(谷胱甘肽S-转移酶)蛋白免疫BALB/C小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合,对杂交瘤细胞及时筛选,阳性孔经三次有限稀释法克隆,成功获得4B3和1H10共2株能稳定传代并分泌抗GST单克隆抗体(McAb)的杂交瘤细胞.2株单克隆抗体腹水间接ELISA效价达10-6,抗体类型均为IgG1,κ链,经Western-blot分析表明,2株单抗与GST-IBV-S1-F融合蛋白及重组GST蛋白均具有特异性反应,验证这2株单抗可用于检测GST融合蛋白的表达情况;用1H10单抗制备的亲和凝胶柱可用于GST融合蛋白的纯化.Two hybridoma cell lines （4B3 and 1H10）secreting monoclonal antibodies （McAbs） against recombinant GST were produced by fusing mouse myelorna cells （SP2/0） with spleen cells from BALB/C immunized by the purified GST protein and subsequently three-time limited dilution method. The titres of ascitic fluids of the two McAbs were up to 10^-6 in indirect ELISA. Isotypes and subclasses of 1H10 and 4B3 all belong to IgG1, κ. Western-blot showed that the two McAbs can specifically react with GST-IBV-S1-F and recombinant GST. The McAb 1H10 has successfully been used in immunoaffinity chromatography of GST fusion protein.

关 键 词：GST 杂交瘤细胞 单克隆抗体 

分 类 号：R392.11[医药卫生—免疫学] Q814[医药卫生—基础医学]