O型口蹄疫病毒单克隆抗体的制备和鉴定

Preparation and characterization of monoclonal antibody against Foot-and-mouth disease virus serotype O

利用O型口蹄疫病毒(FMDV)免疫BALB/c小鼠,取其脾细胞与Sp2/0细胞进行融合,经间接ELISA方法筛选和7次亚克隆后获得2株能稳定分泌单克隆抗体(McAb)的杂交瘤细胞株。经间接ELISA检测,2株杂交瘤细胞株(3E1、7E6)的腹水效价分别为1∶25600、1∶102400。间接免疫荧光试验和特异性试验表明2株McAb可与不同株的O型FMDV反应,而不与其他血清型的FMDV反应。Western blot和叠加试验表明2株单抗可识别O型FMDV结构蛋白VP1上的同一个抗原表位。此特异性McAb的获得将为O型FMDV疫苗的研制和诊断方法的建立奠定基础。

The myeloma cells Sp2/0 were fused with the spleen cells from a BALB/c mice immunized with the FMDV type O （LD50=10^-8.5）. The positive hybridomas clones （3El, 7E6） were obtained after screening by indirect ELISA. The McAbs had a ascites titer of 1：25 600 and 1：102 400, respectively as detected by indirect ELISA. Both McAbs could react with other strains type O FMDV by indirect immunofluorescence assay （IFA）. Western blot and competitive binding assay showed that these McAbs could recognize the same epitope of VP1 protein of FMDV.