本地毛形线虫49 Ku ES重组蛋白单克隆抗体的制备与鉴定  

Production and characterization of monoclonal antibody against 49 Ku ES recombinant protein of Trichinella nativa

作　　者：姜曰晓[1] 路义鑫[1] 王君[1] 宋铭忻[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2007年第11期883-886,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：霍英东青年教师基金(91034);中国博士后科学基金(2004035160);"十一五"国家科技支撑计划重大项目(2006BAD06A09)

摘　　要：采用纯化的本地毛形线虫49KuES重组蛋白免疫BALB／c小鼠，取其脾细胞与SP2／0细胞融合，筛选分泌高滴度McAb的杂交瘤细胞株制备腹水。获得两株稳定分泌抗重组蛋白McAb的杂交瘤细胞株D5和C1。试剂盒鉴定两株McAb均属IgM亚类，其轻链均为K链；间接ELISA检测细胞培养液上清和腹水效价，结果显示，两株细胞培养液上清均达到1：1．28X10^3，腹水效价均达到1：1．28X10^4；Western blot结果表明，获得的两株McAb均能特异性的识别约49Ku处的ES抗原；间接ELISA结果显示两株McAb与日本血吸虫成虫、猪囊尾蚴囊液、猪蛔虫、猪鞭虫抗原均无交叉反应，表明两株McAb特异性良好。BALB/c mice were immunized by recombinant 49 Ku ES protein of Trichinella nat/va. The spleen lymph cells were fused with mouse SP2/0 myeloma cells. Two hybridoma cell lines D5 and C 1 secreting McAb against recombinatnt 49 Ku ES protein were obtained. Both McAbs belonged to IgM with K chainand had an ascites titre of 1：1.28 × 10^3, 1：1.28 × 10^4, respectively. Western blot showed that the McAbs specifically recognized 49 Ku ES antigen and have no cross reaction with adult worm antigen of Schistosoma japonicum, cyst liquid antigen of Cysticercosis cellulosae, polypide antigen of Ascaris suum, polypide antigen of Trichuris suum.

关 键 词：本地毛形线虫 重组蛋白 杂交瘤 单克隆抗体 

分 类 号：S852.7[农业科学—基础兽医学] S852.43[农业科学—兽医学]