禽白血病病毒 p27蛋白单克隆抗体的制备及鉴定

摘要

The purified recombinant protein p27 of avian leukosis virus as an immunogen was used to immu-niz Balb/c mice.By cell culture and monoclonal antibody of dilution method,mAb named 2F3,5C2 and 5C7 were prepared against avian leukosis virus p27 protein by fusing myeloma cell line SP2/0 with spleen lymphocytes of immunized Balb/c mice.By biological characteristic analysis,the number of three strain hybridoma cell chromosomes was about 90.Then immunological identifications were conducted by means of ELISA/Western blot and IFA,the three mAb against p27 protein of ALV possessed high antibody ti-ters and good specificity.The result showed that the three McAbs had good reactogenicity.These three McAbs can be used as a valuable tool for avian leukosis virus epitope analysis,the detection of the virus and Kit research.%以纯化的禽白血病病毒 p27重组蛋白作为免疫原，按常规方法免疫 Balb/c 小鼠，取其脾细胞与 SP2/0细胞融合，通过细胞培养和有限稀释法制备单克隆抗体，最终获得了3株能稳定分泌禽白血病病毒 p27蛋白单克隆抗体的细胞株2F3、5C2和5C7。经生物学特性分析，3株杂交瘤细胞染色体数均为90条左右，通过 ELISA、Western blot 和间接免疫荧光等方法对所获得的3株单克隆抗体进行了鉴定，证明3株单克隆抗体细胞株所分泌的抗体效价高，而且均能够和禽白血病病毒 p27蛋白特异性发生反应。说明该3株单克隆抗体均具有与禽白血病病毒 p27蛋白发生反应的反应原性。3株单克隆抗体的成功获得为禽白血病病毒抗原表位的分析，病毒的检测以及试剂盒的研发奠定了基础。