抗H9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 H9N2 AIV and Identification of its Subtypes

用基因疫苗pcDNA-H9、纯化H9N2亚型AIV和重组噬菌体T4-H9HA免疫Balb/C小鼠,细胞融合后用HI和ELISA方法筛选出5株抗H9亚型AIV HA的特异性单克隆抗体H9-01(2E6)、H9-02(3B1)、H9-03(3H11)、H9-04(1A4)、H9-05(2G9)。各株单抗亚型测定的结果为H9-01、H9-02和H9-05均为IgG2b,H9-03为IgG1,H9-04为IgG2a,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,它们均是HA特异性单克隆抗体。为进一步分析H9N2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。

Balg/C mice were immunized with pcDNA-H9 gene vaccine,purified H9N2 subtype AIV or recombinant bacteriophage T4-H9.Mfter fusion of the splenocytes obtained from the immunized mice with myeloma cells, HI and ELISA screening were done to select hybridoma cell lines,producing five H9 subtype AIV specific monoclonal antibodies H9-01 （2E6）, H9-02（3B1 ）, H9-03（3H11 ）, H9-04（1A4） and H9-05（2G9）. Testing for the subgroup of the munoclonal antibodies revealed that H9-01, H9-02 and H9-05 belonged to the subgroup IgG2b, H9-03 to IgG1, H9-04 to IgG2a. 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. This provided a base for further study on structure and function of H9 subtype AIV to develop the diagnostic kit to detect it.