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目的 获得抗旋毛虫成虫的单克隆抗体 (单抗 )并初步研究其保护性免疫作用。方法 利用杂交瘤技术 ,用旋毛虫成虫抗原免疫的Balb/c小鼠脾细胞与小鼠骨髓瘤细胞SP2 / 0进行细胞融合。用免疫双扩散鉴定单抗亚类 ,免疫印迹鉴定单抗的特异性。小鼠尾静脉接种单抗进行被动免疫保护实验。结果 建立了 1株稳定分泌抗旋毛虫成虫单抗的细胞株。杂交瘤细胞培养上清液效价为1∶6 4 0 0 ,诱生腹水ELISA效价达 1∶2 0 4 80 0。经鉴定单克隆抗体类型为IgM ,可以识别旋毛虫成虫、肌幼虫及新生幼虫 ,相对分子质量约为 4 0 0 0 0蛋白 ,并且不与猪蛔虫、血吸虫、包虫、囊虫抗原发生交叉反应。单抗被动免疫后的减虫率为 5 5 .6 3%。结论 获得 1株具种特异性、保护性的单克隆抗体 ,为筛选能诱导保护性免疫功能的旋毛虫抗原分子提供了有力的工具
Objective To obtain monoclonal antibodies against Trichinella spiralis (mAb) and to study its protective immunity. Methods Balb / c mice immunized with Trichinella spiralis adult spleen cells were fused with mouse myeloma cells SP2 / 0 by hybridoma technique. The double immunodiffusion was used to identify the subtype of McAb and the specificity of the McAb was identified by immunoblotting. Mouse tail vein inoculation of monoclonal antibody for passive immunization experiments. Results A stable cell line secreting monoclonal antibody against Trichinella spiralis was established. The titer of hybridoma cell culture supernatant was 1: 640, and the ascites induced ELISA titer was 1: 240 4 80 0. Monoclonal antibodies identified as IgM, can identify adult Trichinella spiralis, muscle larvae and neonatal larvae, the relative molecular mass of about 40 000 proteins, and does not cross-infection with swine Ascaris, Schistosoma, hydatid, cysticercosis antigen . The worm reduction rate after passive immunization was 55.66%. Conclusion Obtaining a monoclonal antibody with specificity and protection, it provides a powerful tool for screening Trichinella antigen molecules that can induce protective immune function