利巴韦林单克隆抗体的制备及免疫学性能鉴定

Preparation and immunological performance identification of ribavirin monoclonal antibodies

为了获得敏感性高、特异性强的利巴韦林鼠源单克隆抗体。采用戊二醛法将利巴韦林载体蛋白偶联,合成人工完全抗原;紫外扫描、凝胶电泳及动物免疫对人工抗原质量进行鉴定。通过细胞融合、阳性杂交瘤筛选、体内诱生腹水制备单克隆抗体,并对制备的单克隆抗体进行免疫学性能鉴定。结果显示,利巴韦林与载体成功结合在一起,人工抗原制备成功。得到1株可稳定分泌利巴韦林单克隆抗体的细胞株,细胞株分泌的单抗效价在1∶512000以上,对利巴韦林的IC_(50)分别为4.74μg/L,且与金刚烷胺等其他抗病毒类药物及载体蛋白均无交叉反应性。本研究成功制备出灵敏度高、特异性强的利巴韦林单克隆抗体,为利巴韦林免疫学检测方法的建立奠定了坚实的基础。

This study aims to obtain highly sensitive and specific monoclonal antibodies to ribavirin.Ribavirin was coupled with carrier proteins to synthesize artificial complete antigen by glutaraldehyde method.UV scanning,gel electrophoresis and animal immunization were employed to identify quality of the synthesized artificial complete antigens.The monoclonal antibody was prepared by cell fusion,positive hybridoma screening and induction of ascites in vivo,and the immunological characteristics of monoclonal antibodies were identified.The results showed that RBV was coupled to carrier proteins,and the artificial antigens were prepared successfully.One cell line that can stably secrete monoclonal antibodies against ribavirin was obtained,and the titers of the monoclonal antibodies secreted by cell line was above 1∶512000,with IC_(50)values of 4.74μg/L.Furthermore,there were no cross-reactivity with other antiviral drugs such as amantadine and carrier proteins.In this study,monoclonal antibodies of ribavirin with high sensitivity and specificity was synthesized successfully,which lays a solid foundation for the establishment of immunoassay methods of ribavirin.