抗弓形虫MIC3单抗的制备及其在速殖子免疫荧光观察中的应用  

Preparation of monoclone antibodies to Toxoplasma gondii microneme protein 3 and the utilization in tachyzoite immunofluorescent observation

作　　者：刘钊[1] 禹海杰[1] 卓洵辉[1] 周前进[1] 杜爱芳[1] 

机构地区：[1]浙江大学动物科学学院农业部动物疫病病原学与免疫控制重点开放实验室,浙江杭州310029

出　　处：《中国兽医科学》2011年第10期1030-1033,共4页Chinese Veterinary Science

基　　金：浙江省重大科技专项(2009C12029);浙江金华市科技计划重点项目(2010-2-068)

摘　　要：为开展弓形虫速殖子阶段蛋白表达和定位的相关研究,利用弓形虫重组微线体蛋白3(MIC3)抗原研制单克隆抗体,筛选出适合在虫体内用免疫荧光观察该蛋白的单抗分泌株,建立了弓形虫间接免疫荧光观察方法。结果显示,经4轮亚克隆后得到3株稳定分泌抗MIC3单抗的分泌株:4G1、1G12和5D7;经测定,3株MIC3单抗4G1、1G12和5D7的亚型分别为IgG1、IgG2b、IgG2b,效价分别为1∶3 200、1∶12 800、1∶1 600。筛选得到的2株MIC3单抗(4G1和1G12)可用于间接免疫荧光检测MIC3蛋白在虫体中的分布。通过弓形虫阳性感染猪血液样品推片的免疫荧光试验,验证了这3株单抗具有特异性。结果表明,研制的MIC3单抗和虫体荧光观察体系可用于速殖子MIC3蛋白在虫体表达定位和其他研究,也可作为其他速殖子抗原荧光观察的比较对象。To promote research in Toxoplasma gondii tachyzoite protein expression and location,monoclone antibodies（McAbs） to T.gondii microneme protein 3（MIC3） were generated,and screened to set up a system of tachyzoite immunofluorescent observation.In this study,three McAbs strains against MIC3 were subcloned four times.Their titer and subtype were as follows：4G1 was IgG 1,with titer of 1∶3200;5D7 and 1G12 were IgG 2b,with titer of 1∶1600 and 1∶12800,respectively.4G1 and 1G12 were applicable for the immunofluorescent observation of MIC3 in tachyzoite.Anomal-fluorescence was not significant in swine blood immunofluorescent test.The successful immunofluorescent observation system will be utilized in further study for MIC3 expression and location in T.gondii,or as a contrast with other tachyzoite protein.

关 键 词：弓形虫 微线体蛋白3 单克隆抗体 免疫荧光观察 蛋白表达定位 

分 类 号：S852.723[农业科学—基础兽医学]