马传染性贫血病毒基质蛋白p15单克隆抗体的制备  

Preparation of monoclonal antibodies against the matrix protein of equine infectious anemia virus

作　　者：张萍[1,2] 于在江[2] 韩秀娥[3] 王雪峰[2] 朱远茂[2] 周建华[2] 魏萍 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150001 [2]兽医生物技术国家重点实验室／大动物病研究室、中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150001 [3]东北农业大学黑龙江省乳品工业技术开发中心,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2009年第9期730-733,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30771994);十一五重大传染病专项(2008ZX1001-1010)

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。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 plS. 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 1 F12, 1 E2, 1E3, 4B10, 4H7, 5E5, 4G5, respectively. These MAb were shown to specifically react to the pl 5 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-p 15 MAbs would promote further the studies on EIAV and other lentiviruses.

关 键 词：马传染性贫血病毒 基质蛋白p15 单克隆抗体 

分 类 号：S852.4[农业科学—基础兽医学]