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鸭坦布苏病毒E蛋白中和性单克隆杭体的制备
引用本文:姬希文,李雪松,李国新,肖亚莉,颜丕熙,徐大伟,范钊,张七斤,李泽君. 鸭坦布苏病毒E蛋白中和性单克隆杭体的制备[J]. 中国家禽, 2012, 34(4): 25-28,32
作者姓名:姬希文  李雪松  李国新  肖亚莉  颜丕熙  徐大伟  范钊  张七斤  李泽君
作者单位:1. 内蒙古农业大学兽医学院,内蒙古呼和浩特010018;中国农业科学院上海兽医研究所,农业部寄生虫重点开放实验室,上海200241
2. 中国农业科学院上海兽医研究所,农业部寄生虫重点开放实验室,上海200241
3. 内蒙古农业大学兽医学院,内蒙古呼和浩特,010018
4. 中国农业科学院上海兽医研究所,农业部寄生虫重点开放实验室,上海200241;国家家禽工程技术研究中心,上海200241
基金项目:国家自然科学基金面上项目,公益性行业(农业)科研专项经费项目,863计划,国际合作项目
摘    要:本试验利用纯化的鸭坦布苏病毒(Duck Tembusu virus , DTMUV)奉贤株(FX2010)免疫BALB/c小鼠,通过淋巴细胞杂交瘤技术,筛选获得3株能够稳定分泌杭DTMUV的杂交瘤细胞,分别命名为:J1-E5-E4, 7B5和1E5-B6,3株杂交瘤细胞诱导同品系刁、鼠的杭体效价为1:12 800,1:6 400和1:25 600。间接ELISA和IFA结果显示,3株单杭均可以与DTMUV发生特异性反应。进一步研究发现,J1-E5-E4可以与DTMUV-E蛋白结合,使得表达E蛋白的293T细胞显现出特异性的绿色荧光;在病毒中和试验中,J1-E5-E4可以中和DTMUV,使其失去致死鸡胚的能力。这些结果表明,J1-E5-E4是一株针对DTMUV-E蛋白中和表位的单杭,该单杭将在DTMUV诊断和杭原分析中发挥重要作用。

关 键 词:DTMUV  E蛋白  中和  单克隆抗体

Development of Neutralizing Monoclonal Antibody Against Protein E of Duck Tembusu Virus
JI Xiwen,LI Xuesong,LI Guoxin,XIAO Yali,YAN Pixi,XU Dawei,FAN Zhao,ZHANG Qijin,LI Zejun. Development of Neutralizing Monoclonal Antibody Against Protein E of Duck Tembusu Virus[J]. China Poultry, 2012, 34(4): 25-28,32
Authors:JI Xiwen  LI Xuesong  LI Guoxin  XIAO Yali  YAN Pixi  XU Dawei  FAN Zhao  ZHANG Qijin  LI Zejun
Affiliation:2,3(1.College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot,Inner Mongolia 010018;2.Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Shanghai 200241;3.National Poultry Engineering and Technology Research Center,Shanghai 200241)
Abstract:BALB/c mice were immunized with the purified Fengxian strain(FX2010) of duck Tembusu virus(DTMUV).Three monoclonal antibodies(McAb) designated J1-E5-E4,7B5 and 1E5-B6 were obtained by lymphocyte hybridoma technique.The ascites titers of J1-E5-E4,7B5 and 1E5-B6 were 1:12 800,1:6 400 and 1:25 600,respectively.All three McAbs were able to react with DTMUV specifically by indirect ELISA and indirect immunofluorescence assay(IFA).Importantly,J1-E5-E4 was found to bind to protein E of DTMUV specifically in cells and neutralize DTMUV in chicken embryos.The McAb would be useful in clinical diagnosis and antigen analysis of DTMUV.
Keywords:DTMUV  protein E  neutralization  monocolnal antibody
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