抗H3N2亚型猪流感病毒单克隆抗体的制备与鉴定  

Preparation and identification of monoclonal antibodies against H3N2 subtype swine influenza virus

作　　者：孔维[1,2] 乔传玲[1] 陈艳[1] 杨焕良[1] 展小过[1] 辛晓光[1] 陈化兰[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医物技术国家重点实验室/农业部动物流感重点开放实验室,黑龙江哈尔滨150001 [2]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《畜牧与兽医》2010年第9期4-7,共4页Animal Husbandry & Veterinary Medicine

摘　　要：采用纯化的H3N2亚型猪流感病毒(SIV)尿囊液作为免疫原免疫6～8周龄Balb/c小鼠,取免疫小鼠脾细胞与骨髓瘤细胞(SP2/0)融合,用间接ELISA方法筛选分泌抗SIV-H3N2的阳性细胞株,经克隆获得7株亲和力较高的杂交瘤细胞株,分别命名为1C9、2C5、2F10、3D3、4E8、5C7、5D12,用其制备的腹水ELISA效价可达1×106。通过抗体亚型测定,间接免疫荧光试验及免疫印迹试验分析鉴定,该7株单抗均为抗H3N2亚型SIV的特异性单克隆抗体,而且与其他亚型猪流感病毒、猪细小病毒、猪繁殖与呼吸综合征病毒、猪圆环病毒和猪瘟病毒等均无交叉反应,为H3N2亚型SIV的鉴别诊断奠定了基础。Seven monoclonal antibodies(McAbs) against H3N2 subtype swine influenza virus(SIV) were prepared and identified by fusing SP2/0 cell with splenic cells of Balb/c mice that had been immunized with the purified virus antigen.Positive hybridoma cell strains 1C9,2C5,2F10,3D3,4E8,5C7 and 5D12 were determined by ELISA,which showed the highest ELISA titers of 1×106 in ascites.These McAbs were specific to SIV H3N2 by subtype identification,indirect immunofluoescence assay and western-blot.There were no cross reacti...

关 键 词：猪流感病毒 单克隆抗体 ELISA 

分 类 号：S852.65[农业科学—基础兽医学]