抗猪γ-干扰素单克隆抗体的制备和鉴定  

Preparation and characterization of a monoclonal antibody against porcine interferon gamma

作　　者：余斌[1] 张强[1] 闫丽萍[1] 仇华吉[1] 王云峰[1] 陈焕春[2] 童光志[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001 [2]华中农业大学农业微生物学国家重点实验室,湖北武汉430070

出　　处：《中国预防兽医学报》2007年第2期146-149,共4页Chinese Journal of Preventive Veterinary Medicine

摘　　要：将在大肠埃希氏菌中表达的重组猪γ-干扰素(rpIFN-γ)免疫8周龄BALB/c鼠3次后,取脾细胞与SP2/0骨髓瘤细胞按标准程序融合后,以间接ELISA和有限稀释法进行筛选,获得1株能稳定分泌抗猪IFN-γ单克隆抗体的杂交瘤细胞株,该株单克隆抗体命名为B11株。经Western blot分析和间接ELISA检测,B11株能与商品化猪IFN-γ标准品产生特异性反应。间接免疫荧光检测显示B11株能与经PMA刺激的猪外周血单个核细胞产生特异性荧光。抗体亚型鉴定结果表明,B11株为IgG1型,且轻链为κ链。BALB/c mice were immunized three times with purified recombinant porcine interferon-gamma expressed in E.coli. Hybridomas were produced by fusing spleen cells from the immunized BALB/c mouse with SP2/0 myeloma cells according to standard procedure. Positive clones were screened by indirect ELISA and limited dilutions. Finally, a hybridoma which stably secrets specific monoclonal antibody against porcine IFN-γ, was established. The monoclonal antibody, named B11 strain, was shown to strongly react with commercial porcine IFN-γ standard in Western blot and indirect ELISA. The B11 strain also showed strong reactivity in immunofluorescent antibody test on the porcine PBMCs stimulated with PMA. Further characterization revealed that B11 strain belongs to IgG1 isotype, and the light chains are κ chain.

关 键 词：猪Γ-干扰素 单克隆抗体 

分 类 号：Q78[生物学—分子生物学]