抗犬瘟热病毒单克隆抗体杂交瘤细胞株的建立

目的]筛选出稳定分泌抗犬瘟热病毒单克隆抗体的杂交瘤细胞株。方法]用纯化的犬瘟热病毒(CDV)抗原免疫BALB/c小鼠,取脾细胞与SP2/0骨髓瘤细胞融合,经间接ELISA和有限稀释法克隆杂交瘤细胞。研究了杂交瘤细胞的稳定性和染色体数,测定了单克隆抗体的效价并鉴定其特异性和亚型。结果]经反复筛选和亚克隆后,共获得3株能稳定分泌抗CDV单克隆抗体的杂交瘤细胞株。3株单抗对CDV均有较高的特异性,与犬传染性肝炎病毒、犬细小病毒均不反应。3株杂交瘤细胞的腹水效价为1∶(8 000~128 000),细胞培养上清效价为1∶(256~512),染色体数为80~100。3株单克隆抗体重链均属于IgG1,轻链均属于κ链。结论]该研究为研制CDV快速检测试剂盒奠定了基础。

Establishment of Hybridoma Cell Lines Secreting Monoclonal Antibodies against Canine Distemper Virus

Objective] The research aimed to screen out hybridoma cell lines that secreted monoclonal antibodies against canine distemper virus(CDV) stably.Method] BALB/c mice were immunized with the purified CDV antigen.Spleen cells were taken to fuse with SP2/0 myeloma cells and hybridoma cells were cloned by using indirect ELISA and limited dilution method.The stability and chromosome number of hybridoma cells were studied,the titer of monoclonal antibodies were determined and their specificity and subtype were identified.Result] After repeated screening and subcloning,3 hybridoma cell lines that could stably screte monoclonal antibody against CDV were obtained.Monoclonal antibodies in 3 lines all had higher specificity to CDV,which didn't react with infectious canine hepatitis virus and canine parvovirus.The ascites titers of 3 hybridoma cell lines were 1∶(8 000～128 000),the titer of cell culture supernanant was 1∶(256～512) and the chromosome number was 80～100.The heavy chains in monoclonal antibodies of 3 myeloma cell lines belonged to IgG1 and light chains belonged to κ chains.Conclusion] This research laid the foundation for developing the reagent kit for rapid determination of CDV.