论文部分内容阅读
目的制备肠道病毒71型(EV71)单克隆抗体,并进行初步鉴定。方法将RD细胞和Vero细胞培养的EV71原液通过氯化铯超速离心纯化,分别作为免疫抗原和检测抗原,制备单克隆抗体。通过Westernblot分析、间接免疫荧光试验和ELISA法鉴定单抗的特异性。采用间接ELISA法测定单抗的酶标效价,微量细胞培养中和试验测定单抗的中和效价。结果获得1株可稳定分泌抗EV71单克隆抗体的杂交瘤细胞株,其分泌的单抗可与EV71特异性结合,酶标效价为1∶204800,中和效价为1∶28。结论已成功筛选出1株分泌抗EV71的单抗细胞株,为建立EV71疫苗抗原含量测定方法奠定了基础。
Objective To prepare the monoclonal antibody against enterovirus 71 (EV71) and to identify it. METHODS Monoclonal antibodies were prepared by ultracentrifugation of EV71 stock from RD cells and Vero cells by ultracentrifugation using CsCl as immunogen and detection antigen respectively. Western blot analysis, indirect immunofluorescence assay and ELISA were used to identify the specificity of mAb. Indirect ELISA method was used to determine the titer of monoclonal antibody. Neutralization titer of monoclonal antibody was determined by neutralization assay. Results A hybridoma cell line stably secreting anti-EV71 monoclonal antibody was obtained. The secreted mAb could specifically bind to EV71 with a titer of 1:204800 and a neutralizing titer of 1:28. Conclusion A monoclonal antibody against EV71 was successfully screened, which laid the foundation for the establishment of a method for the determination of antigen content of EV71 vaccine.