目的　应用生物传感器对新制备的抗戊型肝炎病毒单克隆抗体 (mAb)的亲和力、Ig亚类 (型 )及抗原结合的动力学进行研究。方法　用HEVORF2区基因工程重组蛋白NE2 ,免疫BALB/c小鼠 ,经杂交瘤技术制备mAb ,采用ELISA、Westernblot和生物传感器鉴定其有关特性。结果获得 12株可稳定分泌抗NE2mAb的杂交瘤细胞系。用ELISA及生物传感器等鉴定各株mAb ,分别为IgM和IgG1、IgG2a,轻链均为κ型。其中在用ELISA法对mAb3F5亚类鉴定过程中 ,发现其与HRP GAMIgG2a和HRP GAMIgM均有反应 ,生物传感器鉴定其为IgM。Westernblot验证各株mAb的特异性 ,同时各株mAb对于不同聚合形式的NE2蛋白的反应性有一定的差别。应用生物传感器测定了 4株mAb的KD 值 ,mAb 8C11为 4 .36× 10 7,mAb8H3为 1.5 3× 10 5,mAb 13D8为 1.2 1× 10 6,mAb 16D7为8.0 3× 10 7。从生物传感器测得的各株mAb与NE2的结合、解离过程中发现 ,mAb 8H3与NE2可快速结合、快速解离 ,其它 3株mAb结合稳定。结论　经多种方法鉴定 ,所获得的 12株杂交瘤细胞分泌的抗体 ,为抗HEVORF2区段的特异性抗体 Objective To study the affinity, Ig subclass (type) and antigen binding kinetics of newly prepared anti-HEV monoclonal antibodies (mAb) using biosensors. Methods BALB / c mice were immunized with the recombinant protein NE2 of HEVORF2 region, and mAb was prepared by hybridoma technology. ELISA, Western blot and biosensor were used to identify mAb. Results Twelve hybridoma cell lines stably secreting anti-NE2 mAb were obtained. The mAbs of each strain were identified by ELISA and biosensor, and they were IgM and IgG1, IgG2a respectively, and the light chains were all κ. Among them, the mAb3F5 subclass identified by ELISA was found to react with both HRP GAMIgG2a and HRP GAMIgM, and the biosensor identified it as IgM. The specificity of each mAb was verified by Western blot, while the mAb of each strain differed somewhat in the reactivity of the different polymeric forms of NE2 protein. KD values ​​of 4 mAbs were determined using a biosensor. The mAb 8C11 was 4.36 × 10 7, the mAb 8H3 was 1.5 3 × 10 5, the mAb 13D8 was 1.2 1 × 10 6, and the mAb 16D7 was 8.0 3 × 10 7. The mAb bound to NE2 was measured by biosensor. During dissociation, mAb 8H3 and NE2 were rapidly bound and rapidly dissociated, and the other 3 mAbs were stable. Conclusions The antibodies secreted by the 12 hybridoma cells identified by various methods were specific antibodies against HEVORF2 segment