抗蓝舌病毒VP7单克隆抗体的制备及其特性

PREPARATION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES AGAINST BLUETONGUE VIRUS VP7

为了提高蓝舌病毒 (BTV)快速免疫诊断方法的灵敏度和特异性 ,对蓝舌病毒VP7特异性单克隆抗体的制备进行了研究。用纯化BTV1颗粒为免疫原 ,以重组BTV13 VP7为筛选抗原建立了能分泌高效价单抗的 6个杂交瘤细胞株 ,抗体滴度最高可达 1∶32 0 0 0 0 0 0。初步结果表明 ,所获单抗与VP7的结合可被不同型BTV参考血清阻断。随后用单抗与重组VP7抗原初步建立竞争抑制ELISA方法 ,对 2 1份血清进行测定 ,结果与间接ELISA检测结果完全符合 ,有可能用于建立我国BTV快速可靠的新型诊断方法 (竞争ELISA)。

In order to improve the sensitivity and specificity of the diagnosis methods for bluetongue virus (BTV),the preparation of monoclonal antibodies against BTV VP7 were conducted.After hyperimmunizing BALB/c mouse using authentic BTV1 virons and screening with recombinant BTV13 VP7 antigen,6 hybridoma cell lines which secret monoclonal antibodies (McAb) were established.The titers of the McAbs were high up to 1∶32 000 000.Further researches showed that the binding of the McAbs to BTV VP7 could be inhibited by BTV reference sera.Then,a competitive inhibition ELISA was established using the VP7 antigen and McAb,primary results showed that the novel competitive ELISA could be in accordance well with indirect ELISA.