猪细小病毒核衣壳VP2蛋白单克隆抗体的制备及部分特性鉴定

Development of monoclonal antibodies against VP2 protein of PPV

以纯化的PPV重组VP2蛋白免疫BALB/c小鼠,通过细胞融合技术,间接ELISA筛选和3次以上细胞克隆,获得了5株稳定分泌抗PPV VP2蛋白单克隆抗体杂交瘤细胞株,分别命名为2D5、1F11、2B4、3C8、1 H3。其染色体平均数均为87～102条,分泌抗体亚类4株为Ig M类,1株为IgG类,Western blot检测表明,5株单抗均识别猪细小病毒VP2蛋白;间接免疫荧光鉴定表明,5株单抗均与PPV全病毒发生反应;间接ELISA鉴定与其他相关病毒的交叉反应性表明,制备的5株单抗不与TEGV、PRV和PEDV反应,表明所制备的抗体与猪细小病毒具有较强的特异性反应。

The BALB/c mice were immunized with purified PPVVP2.Five hybridoma cell lines,named 2D5,F11,2B4,3C8 and 1H3,steadily secreting anti-PPVVP2 McAbs,were obtained.The chromosomal average number of hybridoma cells was 87～102,the 2D5 belonged to IgG2a subtype and others to IgM subtype.Western blot assay indicated that the McAbs can recognize PPVVP2.The result of IFA indicated that five McAbs can react with PPV virus.The result of indirect ELISA demonstrate that five McAbs had no cross reaction with TEGV,PRV and PEDV,indicating high specificity of the McAbs against PPV.