摘要
本研究以纯化原核表达重组蛋白rBovIFN-γ-PET28a-His为免疫原,以重组蛋白为筛选抗原,通过杂交瘤细胞技术,研制出5株稳定分泌rBovIFN-γ单克隆抗体的细胞株,命名为1C11、6E2、7F7、9F5、10C8;腹水效价分别为1∶12 800、1∶6 400、1∶12 800、1∶25 600、1∶12 800。5株单抗中:1C11、7F7、9F5、10C8只与rBovIFN-γ-PET28a-His反应,而不与IFN-α、IFN-β、IL-4、白介素2、鸡IFN-γ等其他细胞因子发生交叉反应;6E2除了与rBovIFN-γ-PET28a-His反应外,还与鸡IFN-γ发生交叉反应。Western blot分析表明,4株单抗均能特异性识别并结合rBovIFN-γ,纯化后的5株单抗的腹水效价分别为1∶6 400、1∶3 200、1∶6 400、1∶12 800、1∶6 400;同时,本研究制备的兔抗rHis-BoIFN-γ多抗效价达1∶25 600。
In this paper, five cell lines of secreted stable monoclonal antibodies named 1Ctl, 6E2, 7F7, 9F5 and 10C8 were prepared through hybridoma cell technology which the recombinant protein rBovIFN-γ-PET28a-His was used as an immunogen and the recombinant protein was used to screen the antigen. The five cell lines＇ ELISA titers of the ascites were 1 ： 12 800, 1 ： 6 400, 1 ： 12 800, 1 ： 25 600 and 1： 12 800, respectively. Among these cell lines, 1Cll, 7F7, 9F5 and 10C8 could only react with rBovIFN-7-PET28a-His recombinant protein, rather than other cytokines such as IFN-α, IFN-β.IL-4.IL-2 and chich- ken -IFN-γ. While, the 6E2 cell lines have a potential cross-react not only with rBovIFN-γ-PET28a-His but also with chichken -IFN-γ. The results of Western-blot showed that all of these four monoclonal antibodies could recognize＇and bind to rBovIFN- γ, specificity, and the ELISA titers of the ascites were 1 ： 6 400, 1 ： 3 200, 1 ： 6 400, 1 ： 12 800 and 1 ： 6 400 respectively after the purification of these 5 monoclonal antibodies. At the same time, the titers of the ascites could be 1 ： 25 600 for the polyelonal antibody to Anti-rabbit rHis-BoIFN-γ made by this research.
出处
《畜牧兽医杂志》
2017年第6期9-11,共3页
Journal of Animal Science and Veterinary Medicine
基金
甘肃省农业科技创新项目(GNCX-2011-43)
