抗H5亚型AIV血凝素单克隆抗体的制备及亚型鉴定  

Preparation of Monoclonal Antibodies against HA of H5N1 AIV and Identification of its Subtypes

作　　者：詹爱军[1] 王新卫[2] 陈枝楠[1] 卢体康[1] 花群义[1] 

机构地区：[1]深圳出入境检验检疫局动植中心 [2]河南农业大学牧医工程学院,郑州450002

出　　处：《中国农学通报》2008年第6期8-11,共4页Chinese Agricultural Science Bulletin

基　　金：国家十一五科技支撑计划项目(2006BAK02A29)

摘　　要：制备抗H5亚型AIV血凝素单克隆抗体;依据淋巴细胞杂交瘤技术,用纯化的H5N1亚型AIV和重组噬菌体T4-H5HA免疫Balb/C小鼠,细胞融合后用ELISA方法筛选特异性单克隆抗体;获得6株抗H5亚型AIVHA的特异性单克隆抗体H5-01(1E6)、H5-02(2C2)、H5-03(2A8)、H5-04(2G6)、H5-05(2E9)和H5-06(3F6)。各株单抗亚型测定的结果为:H5-01、H5-02为IgG1,H5-03、H5-04为IgG2a,H5-05、H5-06为IgG2b,轻链的亚型均为kappa链;经特异性和与同型病毒的反应谱检测,6株均是抗H5亚型AIVHA特异性单克隆抗体,为进一步分析H5N1亚型AIV的结构与功能以及建立监测该亚型AIV的试剂盒奠定了基础。Monoclonal Antibodies against HA of H5N1 AIV were prepared in this article. Balb/C mice were immunized with purified H5N1 subtype AIV or recombinant bacteriophage T4-H5. After fusion of the splenocytes obtained from the immunized mice with myeloma cells,ELISA screening were done to select hybridoma cell lines producing specific monoclonal antibodies. Six H5 subtype AIV specific monoclonal antibodies H5-01（1E6）,H5-02（2C2）,H5-03（2A8）,H5-04（2G6）,H5-05（2E9） and H5-06（3F6） were obtained. Testing for the subgroup of the monoclonal antibodies revealed that H5-01 and H5-02 belonged to IgG1,H5-03 and H5-04 to IgG2a,H5-05 and H5-06 to IgG2b. All their light chains were of the kappa type. Specificity testing and homologous virus reaction spectrum testing proved that they were all HA-specific monoclonal antibodies against H5N1 AIV. This provided a base for further study on structure and function of H5 subtype AIV,and developing the diagnostic kit to detect it.

关 键 词：H5N1亚型AIV 单克隆抗体 亚型 鉴定 

分 类 号：S852.43[农业科学—基础兽医学] S852.65[农业科学—兽医学]