【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。更多还原

【Abstract】 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.更多还原