| 非洲猪瘟病毒K205R蛋白单克隆抗体的制备与鉴定 |
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| 引用本文： | 赵亚茹,韩蓉,杨吉飞,牛庆丽,关贵全,罗建勋,刘光远,殷宏,GRZEGORZ Wozniakowski,刘志杰.非洲猪瘟病毒K205R蛋白单克隆抗体的制备与鉴定[J].中国兽医科学,2019(7):805-811. |
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| 作者姓名： | 赵亚茹 韩蓉 杨吉飞 牛庆丽 关贵全 罗建勋 刘光远 殷宏 GRZEGORZ Wozniakowski 刘志杰 |
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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甘肃省动物寄生虫病重点实验室;Department of Swine Diseases;江苏省动物重要疫病与人兽共患病防控协同创新中心 |
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| 基金项目： | 甘肃省重点研发计划项目(18YF1WA131);农业科技创新工程项目(CAAS-ASTIP-2016-LVRI);国家重点研发计划项目(2017YFD0502300);国家肉牛牦牛产业技术体系项目(CARS-37) |
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| 摘 要： | 将合成的非洲猪瘟病毒K205R基因序列插入pET-28a(+)表达载体,经IPTG诱导表达pET-28a-K205R重组蛋白。用制备的重组抗原免疫BALB/c小鼠,采用化学融合方法融合、筛选,获得了6A4、7F5和7G3三株抗K205R蛋白的杂交瘤细胞株。经鉴定,这3株单克隆抗体(单抗)的亚类均为IgG1型;单抗的腹水效价分别为1∶51200、1∶409600和1∶102400。将这3株杂交瘤细胞连续培养20代,在第20代仍有很高的分泌抗体的能力,表明其稳定性较好。Western-blot检测结果表明,这3株单抗与K205R重组蛋白有良好的反应性和特异性。上述结果表明,本研究制备的这3株杂交瘤细胞株能够稳定分泌抗体,分泌的抗体特异性高,为非洲猪瘟诊断技术的研究奠定了基础。
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| 关 键 词： | 非洲猪瘟 非洲猪瘟病毒 K205R蛋白 单克隆抗体 |
Preparation and identification of monoclonal antibodies against recombinant K205R protein of African swine fever virus |
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| ZHAO Ya-ru,HAN Rong,YANG Ji-fei,NIU Qing-li,GUAN Gui-quan,LUO Jian-xun,LIU Guang-yuan,YIN Hong,LIU Zhi-jie.Preparation and identification of monoclonal antibodies against recombinant K205R protein of African swine fever virus[J].Veterinary Science in China,2019(7):805-811. |
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| Authors: | ZHAO Ya-ru HAN Rong YANG Ji-fei NIU Qing-li GUAN Gui-quan LUO Jian-xun LIU Guang-yuan YIN Hong LIU Zhi-jie |
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| Institution: | (State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou 225009,China) |
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| Abstract: | ZHAO Ya-ru;HAN Rong;YANG Ji-fei;NIU Qing-li;GUAN Gui-quan;LUO Jian-xun;LIU Guang-yuan;YIN Hong;LIU Zhi-jie(State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou 225009,China) |
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| Keywords: | African swine fever African swine fever virus K205R protein monoclonal antibody |
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