犬瘟热病毒单克隆抗体的制备及其生物学特性鉴定

摘要

用纯化后的犬瘟热病毒免疫Balb/c小鼠，采用间接ELISA方法筛选分泌阳性抗体的杂交瘤细胞株，并挑选阳性孔进行亚克隆，用Vero细胞交叉反应试验及间接免疫荧光检测单克隆抗体的特异性。结果获得2株能稳定分泌抗犬瘟热病毒单克隆抗体细胞株，分别命名为062-1、062-2。实验室已有的一株单克隆抗体为6 H8。交叉反应试验及间接免疫荧光染色检测表明，3株单克隆抗体的特异性较好；经过体外连续传代及反复冻存、复苏，杂交瘤细胞均能稳定分泌特异性单克隆抗体。培养上清液及小鼠腹水间接 ELISA效价分别为1∶105和1∶108。相加试验表明，单克隆抗体062-1、062-2具有共同的表位，而单克隆抗体6 H8识别的位点与其他两株不同。%Canine distemper virus was purified and used to immunize Balb/c mice.The hybridoma culture su-pernants were screened by indirect ELISA.The positive cells were subcloned until all clones were positive. The specificity of monoclonal antibodies was confirmed by Vero cross-reaction test and IFA.The results showed that three hybridoma cells secreting anti-CDV McAb were obtained.These two cells were named 062-1,062-2.The lab had one McAb called 6H8.All the three McAbs confirmed by cross-reaction test and IFA were specific against CDV.The positive hybridomas were also able to secrete McAbs stably by series passage in vitro with freezing and recovering.The titers of cell culture supernant and ascites tested by ELISA were 1∶105 and 1∶108 ,respectively.Additivity ELISA results showed that epitopes recognized by 2 McAbs(062-1,062-2)differed from that of by McAb 6H8.