弓形虫SAG1单抗的研制及其初步应用.pdfVIP

.9. 畜牧与兽医 2013 年第45 卷第5 期 弓形虫 SAGl 单抗的研制及其初步应用 蒋蔚，刘迎春，陈永军，安立刚，张花景，王权* (中国农业科学院上海兽医研究所/农业部寄生虫学重点开放实验室，上海 2∞241) 摘要:为开展弓形虫免疫诊断及抗弓形虫感染的相关研究，利用可溶性表达的弓形虫表面抗原SAG1 截短型重组蛋白 (trSAG1 )研制单克隆抗 体，筛选出能够稳定分泌高滴度抗体的杂交瘤细胞株，所分泌的单抗不仅适用于在虫体定位该蛋白，而且具有一定的小鼠体内保护作用。共筛选 出4 株 (6H1 、 6G12 、 7 B4和 7A12) 特异性好的单抗分泌株，经亚型测定， 4 株单抗重链均为 IgG1 ，轻链均为 κ 链。抗体经纯化后测定效价为 1 : 8 仪)() - 1 : 25 600; Western blot 结果显示4 株单抗能够同时识别重组表达及天然弓形虫 SAG1 抗原。筛选得到的2 株抗 SAG1 单抗 (6H1 和 7B4)可用于间接免疫荧光检测 SAG1 蛋白在虫体中的表达定位和分布。小鼠被动免疫保护试验表明， 6H1 和7 B4两株单抗分别或联合使用可稍微 延长被弓形虫感染的小鼠的存活时间。本试验研制的弓形虫 SAG1 单克隆抗体对将来研究弓形虫蛋白定位、弓形虫感染的诊断及防治具有一定的 应用价值。 关键词:弓形虫; SAG1; 单克隆抗体;免疫荧光检测:免疫保护 中图分类号: S852. 7 文献标志码:A 文章编号: 0529-5130( 2013 )05αXl9-05 Preparation of monoclonal antibodies against Toxoplasma gondii and i臼 preliminary application JIANG Wei ，口U Yingchun , CHEN Y ongjun , AN Ligang , ZHANG Huajing , W ANG Quan. (Key Laboratory for Animal Parasitology of Ministry of Agriculture , Shanghai Veterinary Research Institute , Chinese Academy of Agricultural Sciences , Shanghai 200241 , China) Abstract: To develop the detection methods for Toxoplasma gondii , monoclonal antibodies (McAbs) to T. gondii major surface antigen SAGl were generated , and screened and identified. Four strains of hybridoma cells including 6Hl , 6G12 , 784 and 7A12 which had good specificity against T. gondii were obtained. The subtypes of all the four McAbs were determined. The heavy chains were IgG l and the light chains were K-chain. Four McAbs were purified and the titers were from 1 : 8α)() to 1 : 256∞. Western blot analysis demonstrated that all the four McAbs reacted strongly and specifically