抗猪瘟病毒E2蛋白单克隆抗体的制备及生物学特性鉴定  

Preparation of Monoclonal Antibodies against E2 Protein of CSFV and Identification of Their Biological Characteristics

作　　者：聂福平[1,2] 杨俊[1] 李作生[3] 王昱[1,2] 向海洋[1,2] 王国民[1] 肖进文[1] 李应国[1] 

机构地区：[1]重庆出入境检验检疫局/重庆市进出口食品安全工程中心,重庆400020 [2]西南大学动物科技学院,重庆400715 [3]成都军区疾病预防控制中心,云南昆明650032

出　　处：《安徽农业科学》2013年第28期11393-11394,11473,共3页Journal of Anhui Agricultural Sciences

基　　金：国家质检总局科技项目(2010IK023);重庆市科委科技创新能力建设项目(CSTC;2009CB1012)

摘　　要：[目的]制备抗猪瘟病毒（CSFV）的单克隆抗体（MAb）。[方法]以纯化的猪瘟病毒免疫4～6周龄BALB／c雌鼠，取其脾细胞与SP2／O骨髓瘤细胞进行融合，筛选出抗CSFV单克隆抗体杂交瘤细胞株。[结果]经间接ELISA获得3株能稳定分泌抗CSFVE2蛋白的MAb杂交瘤细胞，分别命名为1AS、5F3和4D2。Mab亚型鉴定表明3株MAb均为IgG2b，轻链均为K链。Westernblot结果表明3株Mab均能识别重组E2蛋白。间接免疫荧光试验表明3株Mab均与CSFV呈阳性反应，与猪繁殖与呼吸综合征病毒（PRRSV）、猪圆环病毒（PCV2）、猪传染性胃肠炎病毒（TGEV）呈阴性反应。[结论]该研究为建立CSFV特异性检测方法奠定了基础。[ Objective] The research aimed to prepare monoclonai antibodies （MAb） against classical swine fever virus（CSFV）. [ Method] 4 6 week-old female BALB/c mice were immunized with purified CSFV and its splenic cells were fused with SP2/0 myeloma cells. And the hybridoma cell lines of monoclonal antibodies against classical swine fever virus（CSFV） were screened out. [ Result ] Three strains of MAb hybridoma cell that could stably secrete anti-E2 were obtained by indirect ELISA and named as 1AS, 5F3 and 4D2. The identification results of MAb subtype showed that three strains of MAbs were IgG2b type with Kchain. Western blot results showed that three strains of MAbs could recognize the recombinant E2. Indirect ELISA results showed that three strains of MAbs showed positive reactions with CSFV and showed nega- tive reactions with PRRSV, PCV2 and TGEV. [ Conclusion] The research could lay ihe foundation for establishing the specific detection meth- od of CSFV.

关 键 词：猪瘟病毒 E2蛋白 单克隆抗体 

分 类 号：S852.651[农业科学—基础兽医学]