蓝舌病病毒单克隆抗体的制备及生物学特性鉴定

Preparation and biologic characterization of monoclonal antibody against Blue tongue virus

以纯化的蓝舌病病毒 (BTV) 11型VP7免疫BALB/c小鼠 ,运用淋巴细胞杂交瘤技术 ,借助间接ELISA筛选 ,获得了 3株分泌抗BTV11VP7的群特异性单克隆抗体 (McAb)的杂交瘤细胞株 ,命名为C12 D9、B4 F3 、E1A7。这 3株杂交瘤细胞株连续传代 3个月再经液氮冻存 6个月后复苏培养 ,仍能稳定分泌特异性McAb。 3株McAb均具有ELISA特性和免疫沉淀特性 ,其中C12 D9株上清液的ELISA效价为 1∶2 5 6,腹水的ELISA效价为 10 -6。亚类鉴定 ,C12 D9株为小鼠IgG1,B4F3 、E1A7为IgG2a和IgG2b。琼脂免疫扩散试验和间接ELISA试验表明 ,这 3株均与BTV11型VP7发生特异性反应 ,为BTV群特异性McAb。

Using BALB/c mice immunized with purified VP 7 of Blue tongue virus 11 (BTV 11 ) and lymphocyte hybridoma technique and indirect ELISA screening, three strains of hybridoma cells secreting the group specifical monoclonal antibodies (McAb) to VP 7 of BTV 11 were obtained, which were designated as C 12 D 9 , B 4F 3, and E 1A 7. The three strains of hybridoma cells were able to revived and secrete the specific McAb stablely after to be passaged serially for three months and then to be frozen in liquid nitroger for six months. The three McAb possess features in ELISA and immunoprecipitation teat, and the ELISA titers of C 12 D 9 in the supernatant fluid and in the ascetis were 1∶256 and 10 -6 , respectively. By identification of the subgroup, it was proved that C 12 D 9 was IgG 1, B 4F 3 IgG 2a , and E 1A 7 IgG 2b . It was indicated that the three McAbs can react with VP 7 of BTV 11 specificaly and are BTV group specifc McAbs in agar immunodiffusion test and indirect ELISA.