| 口蹄疫病毒非结构蛋白3A单克隆抗体的制备和鉴定 |
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| 引用本文： | 吴磊,田美娜,卢曾军,付元芳,孙普,刘在新,王建科.口蹄疫病毒非结构蛋白3A单克隆抗体的制备和鉴定[J].中国兽医科学,2010,40(4). |
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| 作者姓名： | 吴磊 田美娜 卢曾军 付元芳 孙普 刘在新 王建科 |
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| 作者单位： | 中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点实验室国家口蹄疫参考实验室,甘肃兰州,730046 |
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| 基金项目： | 国家重点基础研究发展规划(973计划) |
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| 摘 要： | 以原核表达并纯化的口蹄疫病毒(FMDV)非结构蛋白3A免疫BALB/c小鼠,取其脾细胞与小鼠骨髓瘤细胞SP2/0进行融合,用间接ELISA方法筛选杂交瘤细胞,有限稀释法进行亚克隆,获得3株能稳定分泌抗3A蛋白单抗的杂交瘤细胞,分别命名为3H2、4B1、7F5,并对这3株单抗进行了生物学鉴定。结果显示,单抗3H2和4B1为IgG2a亚型,7F5为IgG2b亚型。单抗7F5针对的表位与3H2和4B1两株单抗的表位不一致;这3株单抗均能够与FMDV特异性结合,且单抗4B1与所有3A相关蛋白均反应。稳定性鉴定结果表明,获得的3株杂交瘤细胞均能稳定分泌单克隆抗体。研究结果为进一步探索3A蛋白的结构和功能以及建立诊断方法奠定了基础。
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| 关 键 词： | 口蹄疫病毒 3A蛋白 单克隆抗体 |
Preparation and identification of monoclonal antibodies against nonstructural protein 3A of foot-and-mouth disease virus |
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| WU Lei,TIAN Mei-na,LU Zeng-jun,FU Yuan-fang,SUN Pu,LIU Zai-xin,WANG Jian-ke.Preparation and identification of monoclonal antibodies against nonstructural protein 3A of foot-and-mouth disease virus[J].Veterinary Science in China,2010,40(4). |
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| Authors: | WU Lei TIAN Mei-na LU Zeng-jun FU Yuan-fang SUN Pu LIU Zai-xin WANG Jian-ke |
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| Institution: | WU Lei,TIAN Mei-na,LU Zeng-jun,FU Yuan-fang,SUN Pu,LIU Zai-xin,WANG Jian-ke(National Foot-,-Mouth Disease Reference Laboratory/Key Laboratory of Animal Virology of the Ministry of Agriculture/State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China) |
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| Abstract: | BALB/c mice were immunized with foot-and-mouth disease virus(FMDV) nonstructural protein 3A expressed in Escherichia coli.Splenocytes of the BALB/c mice were fused with myeloma cells SP2/0 for producing hybridoma.Specific antibody was determined by indirect ELISA and positive hybridoma cells were cloned by limiting dilution assay.Three hybridoma cell lines stably secreting monoclonal antibodies(McAbs) against the protein 3A were developed and named 3H2,4B1 and 7F5,respectively.The McAb 7F5 was classified in... |
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| Keywords: | foot-and-mouth disease virus protein 3A monoclonal antibody |
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