抗马传染性贫血病毒Rev蛋白单克隆抗体的制备  

Preparation of monoclonal antibodies against Rev protein of equine infectious anemia virus

作　　者：郭纪珂[1,2] 戈曼[2,3] 张泽力[2,4] 马建 申之义[1] 王晓钧[2] 

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010010 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/大动物病研究室,黑龙江哈尔滨150001 [3]东北农业大学动物医学学院,黑龙江哈尔滨150030 [4]东北林业大学野生动物资源学院,黑龙江哈尔滨150040

出　　处：《中国预防兽医学报》2014年第2期157-159,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(31072113;31222054);兽医生物技术国家重点实验室自主课题(SKLVBP2012;SKLVBP201304);中央级公益性基本科研业务费预算增量项目(2012ZL080)

摘　　要：为制备马传染性贫血病毒(EIAV)蛋白表达调控因子Rev的单克隆抗体(MAb),本研究以原核表达并纯化的重组EIAV Rev为免疫原,免疫BALB/c小鼠,将其脾细胞与SP2/0细胞融合,采用间接ELISA方法筛选阳性克隆,经3次细胞克隆纯化后获得一株能够稳定分泌抗Rev的杂交瘤细胞株。MAb亚型鉴定为IgG2a/κ。Western blot与间接免疫荧光试验结果显示,获得的MAb与真核表达的Rev呈阳性反应。本研究首次制备了抗EIAV Rev的MAb,为Rev的功能研究奠定了基础。To prepare the monoclonal antibody （MAb） against regulator of expression of viral protein （Rev） of equine infectious anemia virus （EIAV）, the SP2/0 cells were fused with the splenic cells from mice immunized with prokaryotically expressed and purified recombinant Rev of EIAV. The hybridoma cells were selected by indirect ELISA and purified by limited dilution method. One hybridoma cell line steadily secreting anti-Rev MAb was obtained which the subtype was IgG2a/κ. The western blot and indirect immunofluorescence assay indicated that the MAb was able to react positively with the native Rev expressed from EIAV. The preparation of MAb against Rev of EIAV provided an useful tool for the further study of Rev function in virus replication.

关 键 词：马传染性贫血病毒 REV 单克隆抗体 

分 类 号：S852.65[农业科学—基础兽医学]