IL-17单克隆抗体的制备与鉴定  

Preparation and Identification of Monoclonal Antibodies against Goat Interleukin-17

作　　者：桑锋锋 高洋[1] 张洪杰[1] 朱伟英[1] 王毅[1] 陈德坤[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100

出　　处：《动物医学进展》2017年第11期52-55,共4页Progress In Veterinary Medicine

基　　金：基金项目:陕北白绒山羊主要疫病防控关键技术研究与示范(2015KTTSNY04-04)

摘　　要：为制备山羊IL-17单克隆抗体,诱导重组菌rIL-17-pET32a-TransB(DE3)表达山羊IL-17重组蛋白(rIL-17),纯化后免疫BALB/c小鼠,取免疫合格的小鼠脾细胞与SP2/0骨髓瘤细胞进行细胞融合,采用间接ELISA方法筛选阳性克隆细胞株。对获得的杂交瘤细胞进行染色体组型分析,并对其分泌的IL-17单克隆抗体进行抗体特异性、抗体类别等分析。结果显示,成功获得了纯化的山羊IL-17原核表达产物;筛选到1株能特异性分泌山羊IL-17单克隆抗体的杂交瘤细胞株-B6,其分泌的单克隆抗体亚型为IgG1,抗体轻链为κ。To prepare IL-17 monoclonal antibody （IL-17 McAb） of goat, the expression vector rIL-17- PET32a-TransB （DE3） was constructed to express the goat IL-17 recombinant protein,then the BALB/c mice were immunized with the purified goat rIL-17.The spleen of immunized mouse was fused with myeloma cell line SP2/0,and the positive hybridoma clones were screened by indirect ELISA.Then chromosome karyotype analysis was performed on hybridoma cells,and the antibody specificity and subtype analyses of the secreted monoclonal antibody of IL-17 were performed.The results showed that the prokaryotic expression product of purified goat IL-17 was obtained successfully, and one hybridoma cell, B6, was screened out,which could produce the monoclonal antibody and specifically identify the eukaryotic expression product of IL-17.The antibody subtype of McAb was IgG1 and the light chain was kappa.

关 键 词：山羊 IL-17单克隆抗体 杂交瘤细胞 酶联免疫吸附试验 

分 类 号：S852.4[农业科学—基础兽医学]