摘要
利用纯化后的传染性胰腺坏死病毒(IPNV VP3)重组蛋白免疫BALB/c小鼠,通过细胞融合技术,采用间接ELISA和有限稀释法筛选杂交瘤细胞,利用染色体鉴定、蛋白印迹和免疫荧光等方法对单克隆抗体进行鉴定,共得到2株能稳定分泌特异性抗体的阳性细胞株,分别命名为2F1、4A7,亚类鉴定2株单抗均为IgG1亚类。ELISA检测其腹水效价,蛋白印迹检测表明获得的2株单抗均能特异性识别IPNV VP3蛋白;间接免疫荧光鉴定表明2株单抗均与IPNV发生反应;间接ELISA检测结果表明2株单抗均不与HSV、SVCV、HRV等病毒反应,与IPNV具有较强的特异性反应。
The BALB/c mice were immunized with purified recombinant protein infectious pancreatic necrosis virus（IPNV） VP3.Two hybridoma cell lines against the VP3 protein were obtained by screening with the indirect ELISA and limiting dilution assay,identification of monoclonal antibodies by western-blot and indirect immunofluorescence assay.These two cell lines named 2F1、4A7 can stably secrete specific antibodies.Two hybridoma cell lines were identified to be IgG1 subtype.Western-blot analysis showed that the two McAbs could react with IPNV VP3 recombinant protein,and the two McAbs could combine with IPNV,which was certified by indirect immunofluorescence,and two McAbs had no reactive capability withVHSV,SVCV and HRV,specificity only for IPNV.
出处
《淡水渔业》
CSCD
北大核心
2013年第3期56-60,共5页
Freshwater Fisheries
基金
国家科技支撑计划(2012BAD25B02)
黑龙江省教育厅科技项目(11541019)
