分泌抗犬细小病毒单克隆抗体杂交瘤细胞株的建立

为制备犬细小病毒(CPV)单克隆抗体,用F81细胞繁殖CPV,病毒液经二乙烯亚胺(BEI)灭活、纯化后免疫BALB/c小鼠,将小鼠脾细胞与SP2/0骨髓瘤细胞进行融合,经筛选获得2株分泌抗CPV单克隆抗体的杂交瘤细胞株,命名为F1和B6.F1和B6分泌的单克隆抗体Ig亚类均为IgG1；以F1和B6制备的腹水,经检测,间接ELISA效价均为1×105,免疫荧光效价分别为1:3200和1∶6400；F1和B6培养上清的抗体中和效价分别为1∶1024和1∶2048,血凝抑制效价分别为1∶2048和1∶4096;免疫印迹(Western-blotting)和抗原表位分析结果显示,F1和B6分泌的单克隆抗体可能分别针对CPV的空间表位和线性表位.2株分泌抗CPV单克隆抗体的杂交瘤细胞株的建立可进一步用于开发CPV特异性诊断制剂和治疗制剂.

Establishment of the Hybridoma Producing Monoclonal Antibodies against Canine Parvovirus

Two hybridomas secreting monoclonal antibodies (mAbs) against Canine Parvovirus (CPV) were established by fusing SP2/0 with spleen cells from BALB/c mice immunized with BEI-kelled and ultracentrifugation purified CPV, naming F1 and B6. The Ig isotypes of mAbs were IgG1. The ELISA antibody titers were 1?105 both and the IFA antibody titers of F1 and B6 were 1:3200 and 1:6400 in ascetic fluid. The neutralization titers were 1:1024 and 1:2048 respectively. The HI titers were 1:2048 and 1:4096 respectively. In the western-blotting and epitope analysis, 2 mAbs reacted with different epitopes of CPV. MAbs reported here could provide valuable tools for further development specific diagnosis and treatment reagent to CPV infection.