【摘要】 将灭活的C型FMDV背部皮下多点注射免疫BABL/c小鼠,取小鼠脾细胞和Sp2/0骨髓瘤细胞在PEG1 500作用下融合,用有限稀释法进行亚克隆.以C型FMDV重组VP1蛋白为抗原,间接ELISA法筛选单抗.以O、Asial、A型FMDV以及猪水疱病病毒为抗原,间接ELISA法检测单抗特异性.获得2株抗C型FMDV的单抗3B3和6D5,亚型鉴定为IgG1、IgG3;腹水效价分别为1∶25 600和1∶51 200;中和效价分别为1∶1 280和1∶640;2株单抗与O、Asial、A型FMDV以及猪水疱病病毒间无交叉反应,表明这2株单抗为高特异性抗C型FMDV单抗.研究结果可为C型FMD的诊断及预防提供基础材料,也可用于病因学及免疫学研究.更多还原

【Abstract】 BABL/c mouse was immunized with serotype C FMDV by subcutaneous multisite injection on its back.The hybridoma was prepared by the fusion of mouse spleen cell and Sp2/0 myeloma cell under the PEG1 500 treatment,and subcloned by limiting dilution.To screen monoclonal antibody,the indirect-ELISA was used with FMDV type C recombinant VP1 protein as an antigen.To detect the specificity of monoclonal antibody,the indirect-ELISA was used with the O,Asial,A serotypes of FMDV and SVDV as antigens.Two strains of monoclonal antibody(3B3,6D5) were obtained.The subtype of 3B3 was IgG1,while the subtype of 3B3 was IgG3.The titers of 3B3 and 6D5 in ascite were 1∶25 600 and 1∶51 200,respectively,while the neutralizing titers were 1∶1 280 and 1∶640,respectively.The two strains had no cross-reaction with the O,Asial,A serotypes of FMDV and SVDV.The two strains were identified as monoclonal antibodies against C serotype of FMDV with high specificity and could be used in diagnostic,prevention,and study of etiology.更多还原