| 抗汉坦病毒NP抗原细胞内抗体的瞬时表达及活性检测 |
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| 引用本文： | 白文涛,徐志凯,张芳琳,罗雯,刘勇,吴兴安,阎岩. 抗汉坦病毒NP抗原细胞内抗体的瞬时表达及活性检测[J]. 细胞与分子免疫学杂志, 2004, 20(6): 705-707 |
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| 作者姓名： | 白文涛 徐志凯 张芳琳 罗雯 刘勇 吴兴安 阎岩 |
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| 作者单位： | 第四军医大学基础部微生物学教研室,陕西,西安,710032 |
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| 基金项目： | 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) |
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| 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 ,为进一步研究细胞内抗体的抗病毒功能奠定基础
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| 关 键 词： | 细胞内免疫 单链抗体 单克隆抗体 汉坦病毒 |
| 文章编号： | 1007-8738(2004)06-0705-03 |
| 修稿时间： | 2004-06-30 |
Transient expression and characterization of intracellular single chain Fv against the nucleocapsid protein of Hantavirus |
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| BAI Wen-tao,XU Zhi-kai,ZHANG Fang-lin,LUO Wen,LIU Yong,WU Xing-an,YAN Yan. Transient expression and characterization of intracellular single chain Fv against the nucleocapsid protein of Hantavirus[J]. Chinese journal of cellular and molecular immunology, 2004, 20(6): 705-707 |
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| Authors: | BAI Wen-tao XU Zhi-kai ZHANG Fang-lin LUO Wen LIU Yong WU Xing-an YAN Yan |
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| Abstract: | 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-Ckappa/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. |
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| Keywords: | intracellular immunization single chain Fv monoclonal antibody Hantavirus |
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