| 核酸免疫制备抗猪流感病毒HA单克隆抗体 |
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| 引用本文： | 李越,王伟,赵颖慧,周长良,臧凤霞,冉多良,孟庆文,陈洪岩.核酸免疫制备抗猪流感病毒HA单克隆抗体[J].中国畜牧兽医,2014,41(7):35-39. |
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| 作者姓名： | 李越 王伟 赵颖慧 周长良 臧凤霞 冉多良 孟庆文 陈洪岩 |
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| 摘 要： | 为制备H1N1猪流感病毒HA蛋白单克隆抗体(monoclonal antibody，McAb)，本试验利用表达猪流感病毒A/Swine/Guangdong/2004(H1N1)毒株HA蛋白的表达质粒pVAX1-HA肌注股内肌免疫BALB/c小鼠，将其脾细胞与骨髓瘤细胞(SP2/0)进行融合；通过间接ELISA方法筛选和有限稀释法克隆，获得9株稳定分泌抗HA单克隆抗体的杂交瘤细胞。中和试验结果显示，单克隆抗体4D5株对H1N1流感病毒起中和作用，该单克隆抗体杂交瘤细胞培养上清的效价为1∶1024。这株单克隆抗体与哈尔滨兽医研究所国家重点实验室保存的H3N2流感病毒、H5N1流感病毒均不发生交叉反应，显示出了很好的H1特异性；间接免疫荧光试验结果显示，这株单克隆抗体能与H1N1流感病毒发生特异性反应。制备的特异性抗HA单克隆抗体为建立H1N1流感病毒免疫学检测方法和单链抗体抗病毒复制研究奠定了基础。
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| 关 键 词： | H1N1流感病毒 血凝素蛋白 单克隆抗体 |
Preparation of Monoclonal Antibody against Swine Influenza Virus HA by Nucleic Acid Immunization |
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| LI Yue,WANG Wei,ZHAO Ying-hui,ZHOU Chang-liang,ZANG Feng-xia,RAN Duo-liang,MENG Qing-wen,CHEN Hong-yan.Preparation of Monoclonal Antibody against Swine Influenza Virus HA by Nucleic Acid Immunization[J].China Animal Husbandry & Veterinary Medicine,2014,41(7):35-39. |
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| Authors: | LI Yue WANG Wei ZHAO Ying-hui ZHOU Chang-liang ZANG Feng-xia RAN Duo-liang MENG Qing-wen CHEN Hong-yan |
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| Abstract: | For the preparation of monoclonal antibody(McAb) against HA protein of swine influenza virus, DNA plasmid bearing HA gene of A/Swine/Guangdong/2004(H1N1) was used to immunize BALB/c mice followed by generation of a McAb using the hybridoma technique. The generated nine McAbs were identified by ELISA. Functional analysis demonstrated the McAb 4D5 neutralized the virus. Furthermore, the McAb 4D5 didn’t have cross-reactivity with H3N2 or H5N1 influenza viruses, showing highly specific to H1N1. Immunofluorescence assay showed that McAb 4D5 was able to detect infection of cells with H1N1. Over all, McAb 4D5 proved specific and highly sensitive for A/Swine/Guangdong/2004(H1N1) HA and could potentially serve as immunodiagnostic tools for the rapid detection of this specific strain of influenza virus and laid the foundation for the study of single chain antibody fragment. |
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| Keywords: | H1N1 influenza virus hemagglutinin protein monoclonal antibody |
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