作　　者： ; ; ; ; ; ; ; ;

机构地区： 四川农业大学动物医学院

出　　处： 《中国兽医科学》 2009年第11期994-998,共5页

摘　　要： 以纯化的猪传染性胃肠炎病毒(TGEV)重组N蛋白免疫BALB/c小鼠,取免疫小鼠的脾细胞与SP2/0骨髓瘤细胞进行融合,通过间接ELISA法对融合细胞进行筛选,采用有限稀释法对阳性细胞进行4次亚克隆,获得了3株能稳定分泌N蛋白单克隆抗体的杂交瘤细胞,分别命名为JY(4)-1、JY(4)-2、JY(4)-3。分别采用体外培养法、诱导腹水法和实体瘤法制备单克隆抗体,并用间接ELISA法检测其效价,结果显示,这3株杂交瘤细胞的细胞培养上清、小鼠腹水和血清的单抗效价均分别为1∶512、1∶105、1∶105,其中以诱导腹水法制备的单抗产量和效价最佳。间接免疫荧光试验结果显示,这3株单抗均能与TGEV感染的ST细胞产生特异性免疫荧光,而不与猪细小病毒、猪生殖与呼吸综合征病毒、猪轮状病毒等发生交叉反应;经鉴定,3株单抗的亚类均为IgG2a,这3株杂交瘤细胞具有良好的遗传稳定性。 The monoclonal antibodies（McAbs） against transmissible gastroenteritis virus（TGEV） N protein were developed by fusing SP2/0 cells with splenocytes of BALB/c mice immunized with the purified recombinant N protein of TGEV.Three hybridoma cells,JY（4）-1,JY（4）-2 and JY（4）-3,were exa-mined by an indirect ELISA and subcloned 4 times by the limited dilution.The titers of 3 hybridoma cells＇culture supernatants,murine ascites and sera were 1∶512,1∶105 or 1∶105 in the indirect ELISA,respectively.The inducing ascites method was the best for producing monoclonal antibodies.The 3 McAbs were in strong reactivity with the ST cells infected with TGEV and no cross-reaction was observed between the McAbs and porcine parvovirus,or porcine reproductive and respiratory syndrome virus or porcine rotavirus in the indirect immunofluorescence assay.The three McAbs,being classified into IgG2a,were highly stable.

关 键 词： 猪传染性胃肠炎病毒 重组 蛋白 单克隆抗体 杂交瘤

领　　域： [农业科学] [农业科学] [农业科学]