【摘要】 目的为了获得针对O型口蹄疫病毒的单克隆抗体,本研究采用差速离心、半饱和硫酸铵沉淀和蔗糖密度梯度离心法提纯O型口蹄疫病毒液,免疫BALB/c小鼠,取脾细胞和SP2/0细胞融合,经ELISA方法筛选和3次亚克隆后获得2株稳定分泌表达的杂交瘤细胞株2C8和5G10,腹水抗体效价分别达到1∶6 400和1∶25 600。经亚类鉴定确定两株单抗均为IgM型,Western-blot、间接ELISA和IFA确定这2株单克隆抗体分别针对O型口蹄疫病毒VP1蛋白不同表位,从而,为研究FMDV的生物学特性和建立快速诊断方法的建立奠定了基础。更多还原

【Abstract】 In order to obtain the monoclonal antibodies(McAb) against foot and mouth disease virus(FMDV),the virus isolate was purified by sucrose density gradient centrifugation and was used to vaccinate the BALB/c mice.After immunization for 3 times,the spleen cells of the mice were used to fuse with the myeloma cell line SP2/0.After screening by ELISA and subcloning for 3 times,two strains of hybridomas 2C8 and 5G10 were obtained.The antibody titers of McAb produced by these two strains of hybidoma in ascites were 1∶6 400 and 1∶25 600,respectively.The results of Western blotting,indirect ELISA and IFA indicated that these McAb could react with different epitopes of VP1 protein of FMDV.Subgroup identification assay confirmed that 2C8 and 5G10 belonged to IgM subgroup.Development of these MAbs will benefit further studies on the biological characteristics of FMDV and establishing rapid diagnostic method.更多还原