摘要
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。
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 -killed and uhracentrifugation purified CPV, naming F1 and B6. The Ig isotypes of mAbs were IgG1. The ELISA antibody titers were 1×10^5 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 of specific diagnosis and treatment reagent to CPV infection.
出处
《中国兽药杂志》
2013年第7期9-12,共4页
Chinese Journal of Veterinary Drug
关键词
犬细小病毒
单克隆抗体
杂交瘤细胞株
canine parvovirus （CPV）
monoclonal antibody （mAb）
hybridoma
