马传染性贫血病毒p26蛋白单克隆抗体的制备

摘要

To prepare the monoclonal antibody(Mab) against equine infectious anemia virus (EIAV), Eight-week-old female BALB/c mice were immunized with purified recombinant p26 protein. Two hybridoma cell lines of El 1 and F8 were produced by using hybridoma technique and the Mabs were identifined and analyzed by indirect ELISA and western blot. The results showed that the Mabs were high specificity to both recombinant p26 and donkey leucocyte attenuated EIAV.%为制备马传染性贫血病毒(EIAV)的单克隆抗体(MAb),本研究以原核表达并纯化的EIAV p26蛋白作为免疫原免疫8周龄的BALB/c小鼠,利用淋巴细胞杂交瘤技术,建立抗EIAV衣壳蛋白p26抗体的杂交瘤细胞.应用重组p26蛋白为抗原建立ELISA筛选方法,获得两株能够稳定分泌特异性MAb的杂交瘤细胞株,分别命名为E11和F8.经试验证明,这些抗p26MAb均能够与感染驴胎皮肤细胞的EIAV和经SDS-PAGE分离的EIAV总蛋白中的p26蛋白结合.这两株p26蛋白MAb的获得将为EIAV的检测及鉴别诊断奠定基础.