鸭肝炎病毒单克隆抗体的研究

Study on Monoclonal Antibodies to Duck Hepatitis Virus

[目的]研制鸭肝炎病毒的单克隆抗体。[方法]将DHV-W株的鸭胚尿囊液经冻融、氯仿处理和超速离心纯化后,免疫BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合,探讨DHV特异性强、敏感性高、简便快速的DHV抗原诊断方法。[结果]经间接ELISA筛选获得1株能稳定分泌DHV的单克隆抗体的杂交瘤细胞株5G8。特性鉴定结果表明该株单抗ELISA效价较高且特异性好,中和特性稍差。杂交瘤细胞冻存6个月和12个月后复苏能稳定分泌特异性抗体。[结论]HDV的McAb成功制备为DHV的快速诊断、筛选特定基因探针等技术奠定了基础。

[Objective] The research aimed to produce monoclonal antibodies to duck hepatitis virus.[Method] BALB/c mice were immunized with allantoid liquid of DHV-W strain in duck embryo that were purified by freezing and melting repeatedly,chloroform treatment and ultracentrifugation.The spleen cells were fused with SP2/0 myeloma cells to discuss rapid and simple DHV diagnosis method with high specificity to DHV and high sensitivity.[Result] A strain of hybridoma cell line 5G8 that could excrete monoclonal antibodies to DHV was obtained by indirect ELISA.The characterization results showed that the monoclonal antibody had high antibody titer,high specificity and poor neutralizing properties.The hybridoma cells that were resuscitated after freezing 6 months and 12 months could secrete stable antibodies against DHV.[Conclusion] The successful preparation of McAb against DHV laid the foundation for the rapid diagnosis of DHV and screening specific gene probe.