| 蚕豆萎蔫病毒单克隆抗体研制 |
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| 引用本文： | 吴建祥,青玲,周雪平,陈正贤,李德葆.蚕豆萎蔫病毒单克隆抗体研制[J].浙江大学学报(农业与生命科学版),1999(2). |
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| 作者姓名： | 吴建祥 青玲 周雪平 陈正贤 李德葆 |
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| 作者单位： | 浙江大学生物技术研究所 |
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| 基金项目： | 国家自然科学基金,浙江农业大学校基金 |
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| 摘 要： | 用蚕豆萎蔫病毒（ＢＢＷＶ）提纯病毒粒子免疫ＢＡＬＢ／ｃ小鼠，取其脾细胞与ＳＰ２／０骨髓瘤细胞融合，用纯化的ＢＢＷＶ病毒对杂交瘤细胞及时筛选，阳性孔经三次有限稀释法克隆，成功获得２株ＢＢ－ＷＶ特异性单克隆抗体的细胞株，分别命名为４Ｄ１１，３Ｇ１２．２株单抗腹水间接ＥＬＩＳＡ效价高达１∶６４００００，抗体类型均为ＩｇＧ１，经Ｗｅｓｔｅｒｎ－ｂｌｏｔｉｎｇ分析表明，２株单抗均是针对ＢＢＷＶ４４．７ｋＤ的外壳蛋白大亚基的特异性抗体．单抗抗原结合位点分析表明４Ｄ１１和３Ｇ１２两单抗作用于同一抗原决定簇
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| 关 键 词： | 蚕豆 蚕豆萎蔫病毒 单克隆抗体 |
Production of monoclonal antibodies against broad bean wilt virus |
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| WU Jian xiang,QING Ling,ZHOU Xue ping,CHEN Zheng xian,LI De bao.Production of monoclonal antibodies against broad bean wilt virus[J].Journal of Zhejiang University(Agriculture & Life Sciences),1999(2). |
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| Authors: | WU Jian xiang QING Ling ZHOU Xue ping CHEN Zheng xian LI De bao |
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| Abstract: | Two hybridoma cell line, secreting monoclonal antibodies (MAbs) against broad bean wilt virus (BBWV) were produced by fusing mouse myeloma cells (SP 2/0) with spleen cells from BAL B/c immunized with BBWV purified virus particles and subsequently three time limited dilution method The titres of ascitic fluids of the both McAbs were 1:640 000 when measured by indirect ELISA In agarose gel immunodiffusion test, it showed that the two MAbs contained same isotype of murine antibodies, IgG1 The result of Western blotting showed that both the McAbs recognized with the 44 7 kD a large coat protein subunit of BBWV Antigen binding epitopes analysis showed the two McAbs reacted with the some antigen determinant |
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| Keywords: | broad bean broad bean wilt virus monoclonal antibodies |
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