H9N2禽流感病毒核蛋白NP单克隆抗体的制备

Preparation of monoclonal antibody against NP protein of H9N2 Avian influenza virus

为了了解抗H9N2禽流感病毒(AIV)单克隆抗体的确切用途,试验用纯化后的H9N2禽流感病毒免疫Balb/c小鼠,经细胞融合后再进行筛选。结果表明:获得2株能够稳定分泌抗H9N2 AIV的杂交瘤细胞株,分别命名为1E7和3D6,2株单克隆抗体的腹水效价分别为1∶25 600和1∶12 800;应用间接ELISA和间接免疫荧光检测显示,2株单克隆抗体均能与H9N2亚型AIV发生反应;进一步研究发现,3D6可以与H9N2 AIV中的NP蛋白结合,使得表达NP蛋白的MDCK细胞显现出绿色荧光。说明3D6是一株针对H9N2 AIV NP蛋白的单克隆抗体。

To understand the exact purpose of the monoclonat antibody against H9 subtype of Avain influenza virus, the Balb/c mice were iuanu-nized with the purified H9N2 Avian influenza virus （AIV） , and then the monoclonal antibodies （McAbs） were screened through cell fusion. The results showed that the two hybridoma cell lines secreting McAbs against H9N2 AIV were obtained and named as 1 E7 and 3D6. The ashcites titers of the two McAbs were 1：25 600 and 1：12 800, respectively. It was found that all the two McAbs could react with H9 subtype AIV by u-sing indirect ELISA and indirect immunofluorescence assay （IFA）. A further study flmnd that 3D6 could combine with the NP protein of H9N2 AIV, and the green fluorescence was shown in Madin - Darby canine kidney （MDCK） cells expressing the NP protein. The results indicate that 3D6 is a monoclonal antibody against the NP protein of H9N2 AIV.