| 抗AAV2单克隆抗体的制备及特性研究 |
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| 引用本文： | 谭淑萍,袁振华,田文洪,董小岩,吴小兵.抗AAV2单克隆抗体的制备及特性研究[J].病毒学报,2009(4). |
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| 作者姓名： | 谭淑萍 袁振华 田文洪 董小岩 吴小兵 |
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| 作者单位： | 中国科学院研究生院;中国疾病预防控制中心病毒病预防控制所病毒基因工程国家重点实验室;复旦大学生命科学学院遗传学研究所遗传工程国家重点实验室;北京五加和分子医学研究所有限公司; |
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| 基金项目： | 973计划课题(2004CB518806);;国家自然科学基金课题(30370067) |
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| 摘 要： | 以纯化的重组AAV2病毒颗粒为抗原免疫小鼠,获得7株稳定分泌抗AAV2衣壳蛋白的单克隆抗体杂交瘤细胞株,其中B10和G4两株单克隆抗体具有中和活性,抗体亚型分别为IgG1和IgG2a型。对这两株单克隆抗体与rAAV病毒结合的特性进行了研究。单克隆抗体B10和G4对rAAV2病毒颗粒的结合均具有良好的血清型特异性,并且这种特异结合作用不被肝素阻断。这两株抗体都不阻断AAV2病毒与敏感细胞的结合,提示它们与病毒颗粒的结合位点都不处于AAV2病毒与主要受体结合的部位内。Western blotting检测结果显示,B10与AAV2的三种衣壳蛋白VP1、VP2和VP3均能结合,而G4不能与AAV2的这三种衣壳蛋白结合。这说明B10与AAV2结合的位点位于衣壳蛋白VP1、VP2和VP3的重叠部分处并且可能是线性表位,而G4则可能是针对AAV2病毒颗粒构象表位的抗体。这两种结合特性不同的单克隆抗体为研究AAV2病毒颗粒的表面特性和感染特性提供有用的工具。
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| 关 键 词： | 2型腺相关病毒 单克隆抗体 中和抗体 构象表位 |
The Preparation of Monoclonal Antibodies against AAV2 and Study on Their Characterizations |
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| TAN Shu-ping,YUAN Zhen-hua,TIAN Wen-hong,DONG Xiao-yan,WU Xiao-bing.The Preparation of Monoclonal Antibodies against AAV2 and Study on Their Characterizations[J].Chinese Journal of Virology,2009(4). |
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| Authors: | TAN Shu-ping YUAN Zhen-hua TIAN Wen-hong DONG Xiao-yan WU Xiao-bing |
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| Institution: | 1.Graduate University of Chinese Academy of Sciences;Beijing 100049;China;2.National Key laboratory of Molecular Virology and Genetic Engineering;Institute of Viral Disease Control and Prevention;CDC;Beijing 100052;3.National Key laboratory of Genetic Engineering;Institute of Genetics;School of Life Science;Fudan University;Shanghai 200433;4.Beijing FivePlus Molecular Medicine Institute Co.Ltd;Beijing 100176;China |
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| Abstract: | 7 strains of stable cell lines secreting monoclonal antibodies against AAV2 capsids were obtained by immunizing BALB/C mice with highly purified recombinant adeno-associated virus.Among them,the monoclonal antibodies B10 and G4 had neutralizing activity,and their subtypes were IgG1 and IgG2a,respectively.The binding characterizations of the two neutralizing antibodies were studied.Both B10 and G4 showed serotype specific binding activities to rAAV2 virus particles other than AAV1,AAV5,and AAV8,and the bindi... |
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| Keywords: | Adeno-associated virus type 2 monoclonal antibody neutralize antibody conformational epitope |
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