摘要
以H5亚型禽流感病毒采用长程免疫法免疫的BALB/c小鼠,取4次免疫后的脾细胞与SP2/0骨髓瘤细胞用PEG-1500进行细胞融合,融合率为65.8%.建立间接ELISA,用于检测杂交瘤细胞上清中的单克隆抗体.初检阳性率为14.2%,经2次亚克隆反复筛选获得了2株能稳定分泌抗禽流感病毒单克隆抗体的杂交瘤细胞株,命名为5A6,5G7.ELISA证实所获得的单抗是特异性针对H5和H9亚型禽流感病毒的共同表位,5A6,5G7经反复冻存、复苏及多次传代,仍能分泌高效价抗体,乳胶凝集试验证实所分泌抗体均属IgG1,κ型.
The BALB/c mice were immunized four times with the inactive purified subtype H5 avian influenza virus （AIV-H5）, then the spleen cells of the hyperimmunized mouse and SP2/0 cells were fused with PEG-1500. The cell fusion rate of 65.8 % was obtained. An indirect enzyme-linked immunosorbent assay （ELISA） was developed to detect monoclonal antibodies （McAbs） secreted by hybridoma cell lines. And a positive rate of 14.2 % was obtained. Two hybridoma cell lines secreting the monoclonal antibodies to AIV, named 5A6, 5G7 were developed by two subclones. The specificity of McAb was identified by means of indirect ELISA. After repeated frozen storage,thawed revival and generation, they were still steadily producing high titer of McAbs. The results of latex agglutination test showed that the antibodies secreted by 5A6 and 5G7 cells belong to IgG1, κisotype. The McAbs prepared would be very useful in diagnosis and control of avian influenza.
出处
《重庆工学院学报》
2006年第11期11-14,19,共5页
Journal of Chongqing Institute of Technology
基金
重庆市防治高致病性禽流感应急专项课题资助项目(渝科发字【2004】9号)
