类禽型H1N1猪流感病毒HA蛋白单克隆抗体的制备及特性分析

Preparation and characteristics of monoclonal antibodies against HA protein of the avian-like H1N1 swine influenza virus

为获得类禽型H1N1猪流感病毒（SIV）血凝素（HA）蛋白的单克隆抗体（McAb）,选取病毒A/swine/Zhejiang/245/2013（H1N1）,经增殖、纯化后作为免疫原,3次免疫BALB/c小鼠,取其脾与骨髓瘤细胞（SP2/0）进行融合,利用ELISA方法进行筛选,克隆得到4株杂交瘤细胞株,分别命名为5H7、2H3、2G4、8D7。4株单克隆抗体的腹水ELISA效价为1×10^9以上,HI效价高于8 log2。Western-blot试验与间接免疫荧光试验结果表明,4株McAbs均可与病毒HA蛋白发生特异性的反应。本研究将为进一步建立H1亚型SIV的相关检测方法及开展HA蛋白表位的研究奠定基础。

The purified whole-virus protein derived from A/swine/Zhejiang/245/2013（ZJ245） was chosen to immunize BALB/c mice to prepare the monoclonal antibody（Mc Ab） against hemagglutinin（HA）protein of an avian-like H1N1 swine influenza virus（SIV）.The splenic cells of the mice were fused with the SP2/0 cells after three times of immunization.After screening by indirect ELISA,four hybridomas secreting Mc Abs against HA protein of the ZJ245 were obtained,which was designated as 5H7,2H3,2G4 and 8D7,respectively.ELISA titers of the four Mc Abs were higher than 1×10~9 and the HI titers were higher than 8 log2 in ascites.The results of Western-blot and indirect immunofluorescence assay showed that these four Mc Abs could specifically react with the HA protein of ZJ245.The preparation of these Mc Abs will provide the basis for the development of the diagnostic method and identification of HA protein epitopes of H1 subtype SIVs.