猪圆环病毒衣壳蛋白单克隆抗体的制备

Preparation and identification of two monoclonal antibodies against the capsid protein of Porcine circovirus

以大肠杆菌表达的猪圆环病毒2型(PCV2)重组衣壳蛋白(GST-CAP2)为抗原,以一定的免疫程序接种BALB/c小鼠后,取其脾细胞与骨髓瘤细胞SP2/0按常规杂交瘤技术进行融合。以GST-CAP2以及猪圆环病毒1型(PCV1)重组衣壳蛋白(GST-CAP1)为包被抗原,经间接ELISA筛选获得2株能稳定分泌针对PCV的单克隆抗体的杂交瘤细胞株(M1和G1)。通过dot-ELISA、Western-blotting以及间接免疫荧光(IFA)技术证明G1株分泌的单抗同时识别PCV1和PCV2,而M1株分泌的单抗则特异性针对PCV2。该单抗的制备将为进一步建立快速检测PCV的方法奠定基础。

To prepare a novel specific mouse anti-PCV monoclonal antibody,female BALB/c mice of 8 weeks old were immunized with the recombinant capsid protein of PCV2 expressed in Escherichia coli BL21.Following the last immunization,the spleen B cells of the mice were fused with SP2/0 myeloma cells.A recombinant capsid protein-based indirect-ELISA was used to screen out the hybridoma cells,and two interested hybridoma cell lines were obtained（designed as M1 and G1）.The data of Western-blotting,dotELISA and indirect immunofluorescent assay confirmed that the G1 monoclonal antibody can recognize both PCV1 and PCV2,while the M1 monoclonal antibody is distinctively specific to PCV2.