【摘要】 为制备抗马传染性贫病毒(EIAV)的单克隆抗体(MAb),本研究用纯化的重组EIAV基质蛋白作为免疫原免疫BALB/c小鼠,采用淋巴细胞杂交瘤技术,建立了抗EIAV基质蛋白p15抗体的杂交瘤细胞。应用重组p15和EIAV总蛋白为抗原建立的ELISA以及EIAV感染细胞的间接免疫荧光法,对杂交瘤细胞进行了有限稀释法筛选,获得了7株稳定分泌抗p15抗体的杂交瘤细胞株,分别命名为:1F12、1E2、1E3、4B10、4H7、5E5、4G5。这些抗p15 MAb均能与感染驴胎皮肤细胞的EIAV和经SDS-PAGE分离的EIAV总蛋白中的相应蛋白结合。抗体效价叠加实验表明,这7株MAb中含有至少3种识别不同抗原决定簇的抗体。这些p15 MAb的获得有助于对EIAV和慢病毒的深入研究。更多还原

【Abstract】 A hybridoma library specific to the matrix protein(p15) of equine infectious anemia virus(EIAV) was established by fusing the myeloma cells with spleen B cells from BALB/c mice immunized with recombinant p15.Hybridoma were screened for monoclonal antibodies(MAb) by ELISA and indirect fluorescent assay(IFA) using recombinant p15 and total EIAV protein as antigens.Seven p15 MAb clones were obtained and designated 1F12,1E2,1E3,4B10,4H7,5E5,4G5,respectively.These MAb were shown to specifically react to the p15 on EIAV infected FDD cells and to the total EIAV proteins separated by SDS-PAGE.Analysis of synergistic effects showed that these MAbs contained at least three types of antibodies,which identify individual p15 epitops.The development of these anti-p15 MAbs would promote further the studies on EIAV and other lentiviruses.更多还原