抗汉坦病毒NP抗原细胞内抗体的瞬时表达及活性检测  

Transient expression and characterization of intracellular single chain Fv (against) the nucleocapsid protein of Hantavirus

作　　者：白文涛[1] 徐志凯[1] 张芳琳[1] 罗雯[1] 刘勇[1] 吴兴安[1] 阎岩[1] 

机构地区：[1]第四军医大学基础部微生物学教研室,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2004年第6期705-707,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目 (No.39970 70 2 ;No .30 1 70 0 4 8) ;军队"十五"青年基金资助项目 (0 1Q1 1 5)

摘　　要：目的 :在真核细胞中表达抗汉坦病毒NP抗原鼠源性单克隆抗体 (mAb) 1A8的scFv ,并检测表达产物的活性。方法 :将真核表达载体 1A8 scFv Cκ/pCI neo用脂质体转染COS 7细胞 ,用间接免疫荧光和免疫共沉淀法检测瞬时表达产物的免疫学活性。结果 :间接免疫荧光的结果表明 ,约 1%细胞胞质中出现弥散荧光 ,在免疫共沉淀 /免疫印记中 ,表达的抗体片段可与汉坦病毒的NP抗原特异性结合。结论 :细胞内表达的1A8 scFv Cκ可与NP抗原特异性结合 。AIM: To transiently express an intracellular single chain Fv of monoclonal antibody 1A8 against nucleocapsid protein of Hantavirus and characterize the immunological activities of the expressed products. METHODS: COS-7 cells were transfected with mammalian expression vector 1A8-scFv-Cκ/pCI-neo via lipofectin. The expressed product was identified by indirect immunofluorescence and immunoprecipitation. RESULTS: A diffuse pattern fluorescence was observed in less than 1% cytoplasm of transfected COS-7 cells. The binding of intracellular antibody fragments to NP antigen was confirmed by immunoprecipitation analysis. CONCLUSION: Transiently expressed single chain intrabodies can effectively target NP antigen in the cytoplasm. The present study may provide a new approach for treatment of Hantavirus.

关 键 词：细胞内免疫 单链抗体 单克隆抗体 汉坦病毒 

分 类 号：R373.32[医药卫生—病原生物学]