J亚群禽白血病病毒gp85蛋白单克隆抗体的制备和鉴定  

Preparation and Identification of Monoclonal Antibody Against gp85 of Avian Leukosis Virus Subgroup J

作　　者：刘丽娜[1] 邵华斌[1] 栗绍文[2] 杨峻[1] 程国富[2] 胡薛英[2] 罗青平[1] 

机构地区：[1]湖北省农科院畜牧兽医研究所动物胚胎工程及分子育种湖北省重点实验室,湖北武汉430064 [2]华中农业大学动物医学院,湖北武汉430070

出　　处：《中国家禽》2014年第23期21-24,共4页China Poultry

基　　金：现代农业产业技术体系建设专项资金(CARS-42-G11);农业部公益性行业(农业)科研专项(201303044);湖北省科技计划研究与开发项目(2010BBB008)

摘　　要：制备J亚群禽白血病病毒(ALV-J)gp85蛋白单克隆抗体,采用本实验室表达的ALV-J gp85蛋白作为免疫原,按照常规方法免疫BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞进行融合,利用获得的杂交瘤细胞制备腹水,用辛酸-硫酸铵沉淀法对腹水进行纯化。结果获得了5株杂交瘤细胞:1B9、1G4、3A1、3D12、5G5。杂交瘤细胞上清抗体效价均在27以上,腹水抗体效价均在29以上。Western blot和间接免疫荧光试验表明,单克隆抗体可以与ALV-J gp85蛋白发生特异性反应。本研究制备的单克隆抗体为ALV-J抗原诊断试剂盒的开发奠定了基础。To develop monoclonal antibodies （MAbs）against avian leukosis virus subgroup J （ALV-J）gp85,a panel of MAbs was generated by fusing SP2/0 with spleen cells from BALB/c mice immunized with purified protein gp85. The obtained hybridoma cells were used to prepare the ascites in mice. The ascites were purified using octanoic and ammonium sulfate precipitation. Five of hybridomas cell strains stably secreted MAbs against ALV-J gp85 protein were named as 1B9,1G4,3A1,3D12 and 5G5. These MAbs showed high titers with over 27 of cell supernatant and over 109 of ascite and specific to ALV-J gp85 in western blot and IFA. The MAbs gp85 against ALV-J acquired in this study would be very useful for diagnosis of ALV-J.

关 键 词：J亚群禽白血病 gp85蛋白 单克隆抗体 

分 类 号：S852.65[农业科学—基础兽医学]