【摘要】 为获得抗鹅细小病毒(GPV)VP1蛋白的单克隆抗体,以纯化的VP1蛋白为免疫原,免疫4～6周龄的雌性BALB/c鼠,经4次免疫后,取其脾细胞与SP2/0骨髓瘤细胞进行融合,共获得5株能稳定分泌特异性抗体的阳性细胞株。Western-blot结果表明,这5株杂交瘤细胞分泌的抗体均能特异地与纯化后的GPV-VP1重组蛋白发生反应,单克隆抗体亚型鉴定结果表明,3株单抗为IgG1亚型,1株单抗为IgG2b亚型,1株单抗为IgA亚型,轻链均为κ链。用单克隆抗体对分段表达的VP1蛋白进行免疫印迹分析,结果鉴定出3个抗原表位识别区,分别位于VP1的第92～123、92～145和111～145aa。证实,这5株杂交瘤细胞在体外长期培养能稳定地分泌抗GPVVP1的单克隆抗体。更多还原

【Abstract】 Female BALB/c mice of 4-6 weeks-old were immunized with the purified recombinant goose parvovirus(GPV) fusion protein.After the 3rd-immunization,the spleen cells of the immunized mice were fused with mouse myeloma cells SP2/0.Five positive hybridoma cell lines stably secreting monoclonal antibodies(mAbs) were selected by 3 cycles of limited dilutions.All mAbs recognized the purified GPV-VP1 protein in Western-blot,indicating that all mAbs possessed good specificity.Three mAbs were classified into IgG1 subtype,one mAb was classified into IgG2b subtype and the other one mAb was classified into IgA subtype.Light chains of all mAbs were kappa.Taking advantage of the expressed VP1 fragments,the epitopes corresponding to the mAbs were analyzed.The results showed that the three mAbs recognized the same epitope,which was located in 91-145aa.The other two epitopes were identified and they were located in 91-145 and 111-145 aa,respectively.更多还原