GⅡ型诺如病毒衣壳蛋白抗原表位预测及单克隆抗体的制备

陈莉萍 纪蕾 吴晓芳 徐德顺 韩建康

陈莉萍, 纪蕾, 吴晓芳, 徐德顺, 韩建康. GⅡ型诺如病毒衣壳蛋白抗原表位预测及单克隆抗体的制备[J]. 疾病监测, 2014, 29(1): 66-70. doi: 10.3784/j.issn.1003-9961.2014.01.018
引用本文: 陈莉萍, 纪蕾, 吴晓芳, 徐德顺, 韩建康. GⅡ型诺如病毒衣壳蛋白抗原表位预测及单克隆抗体的制备[J]. 疾病监测, 2014, 29(1): 66-70. doi: 10.3784/j.issn.1003-9961.2014.01.018
CHEN Li-ping, JI Lei, WU Xiao-fang, XU De-shun, HAN Jian-kang. Capsid protein antigen epitope prediction and monoclonal antibody preparation for Norovirus GⅡ[J]. Disease Surveillance, 2014, 29(1): 66-70. doi: 10.3784/j.issn.1003-9961.2014.01.018
Citation: CHEN Li-ping, JI Lei, WU Xiao-fang, XU De-shun, HAN Jian-kang. Capsid protein antigen epitope prediction and monoclonal antibody preparation for Norovirus GⅡ[J]. Disease Surveillance, 2014, 29(1): 66-70. doi: 10.3784/j.issn.1003-9961.2014.01.018

GⅡ型诺如病毒衣壳蛋白抗原表位预测及单克隆抗体的制备

doi: 10.3784/j.issn.1003-9961.2014.01.018

Capsid protein antigen epitope prediction and monoclonal antibody preparation for Norovirus GⅡ

  • 摘要: 目的 原核表达诺如病毒衣壳蛋白VP1,制备抗诺如病毒衣壳蛋白VP1的单克隆抗体,并研究其抗原表位特性。方法 PCR扩增GⅡ型湖州株诺如病毒VP1蛋白基因,将目的基因克隆至载体pET28a(+),转化至大肠埃希菌原核表达。纯化表达产物免疫BALB/c小鼠,将成功免疫的小鼠脾细胞与骨髓瘤SP2/0细胞融合。用Modeller9.9为诺如病毒GⅡ型湖州株VP1蛋白构建三维模型,构建好的三维模型提交到SEPPA在线B-cell 空间表位预测软件,对GⅡ型湖州株VP1蛋白进行空间表位预测。用预测的表位序列来筛选阳性克隆,得到对应这些表位的单克隆抗体。结果 成功构建重组表达载体pET28a(+)-Noro-VP1并在大肠埃希菌中获得表达,重组蛋白免疫的BALB/c小鼠的脾细胞与SP2/0细胞融合,用预测的抗原表位多肽筛选,获得10株杂交瘤细胞株。与重组蛋白的ELISA结果显示1E8、1P10、2C15和2L16四株反应性最强,其他几株反应性较弱。Western blotting结果显示只有2K10、1K16、1E8三株能与重组蛋白很好的结合。结论 成功制备了抗GⅡ型湖州株诺如病毒VP1蛋白的单克隆抗体,抗原表位位于衣壳蛋白的P2区,为制备诺如病毒快速免疫诊断试剂盒及抗原表位的研究提供基础。
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出版历程
  • 收稿日期:  2013-01-10
  • 刊出日期:  2014-01-20

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