›› 2011, Vol. 38 ›› Issue (9): 190-193.

• 流行病防治 • 上一篇    下一篇

H1N1猪流感病毒血凝素蛋白抗原表位的筛选及其抗原性分析

唐连飞, 易征璇, 朱中武, 朱金国, 陈盼, 孟芳, 禹思宇, 欧阳振宇, 姜金林   

  1. 湖南出入境检验检疫局检验检疫技术中心,湖南长沙 410004
  • 收稿日期:2011-01-13 修回日期:1900-01-01 出版日期:2011-09-20 发布日期:2011-09-20

Epitope Screening and Antigenicity Analyzing of Hemagglutinin of H1N1 Swine Influenza Virus

TANG Lian-fei, YI Zheng-xuan, ZHU Zhong-wu, ZHU Jin-guo, CHEN Pan, MENG Fang, YU Si-yu, OUYANG Zhen-yu, JIANG Jin-lin   

  1. Inspection and Quarantine Technical Centre of Hunan Inspection and Quarantine Bureau, Changsha 410004, China
  • Received:2011-01-13 Revised:1900-01-01 Online:2011-09-20 Published:2011-09-20

摘要: 本研究旨在对H1N1猪流感病毒血凝素蛋白抗原分子的模拟表位进行分离鉴定并分析其抗原性。用抗H1N1猪流感病毒血凝素蛋白小鼠血清IgG对噬菌体随机12肽库进行筛选,3轮亲和筛选后,特异性噬菌体得到了有效富集;对随机挑选的47个噬菌体克隆用ELISA进行鉴定,其中40个为阳性;对40个阳性克隆测序得到3种不同氨基酸序列;用Western blotting对获得的3个不同序列的噬菌体克隆进行抗原性分析,显示这3种噬菌体插入短肽能和H1N1猪流感病毒感染小鼠血清特异性结合。结果表明,本试验获得了3种H1N1猪流感病毒血凝素蛋白模拟抗原表位,它们都具有明显的抗原性,为H1N1猪流感病毒的疫苗研究和诊断奠定了基础。

关键词: 噬菌体肽库; 抗原表位; 猪流感病毒; 血凝素蛋白

Abstract: In an attempt to isolate and identify the epitopes on hemagglutinin(HA) of H1N1 swine influenza virus and to test their antigenicity,polyclonal mouse antisera IgG against recombinant HA was prepared to screen a 12-mer random peptide library. After three rounds of biopanning a phage enrichment was resulted. Forty of the 47 phage clones selected randomly showed to be positive by phage-ELISA and shared 3 amino acid sequences. The result of Western blotting showed that the 3 peptides could bind specially with the polyclonal mouse antisera IgG against H1N1 swine influenza virus indicated that the 3 peptides was antigenicity. In this paper,three antigenic epitopes on HA were obtained and established the foundations for the further research of vaccine and diagnoses of H1N1 swine influenza virus.

Key words: phage displayed peptide library; antigen epitope; swine influenza virus; hemagglutinin

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