黄瓜绿斑驳花叶病毒单克隆抗体的研制

摘要

将纯化的黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virus,CCMMV)制剂免疫BALB/c小鼠,最后一次免疫后第3天取其脾细胞与SP2/0细胞融合,经采用选择性培养基、有限稀释法克隆和间接ELISA方法进行筛选,成功获得了3株分泌CGMMV特异性单克隆抗体的杂交瘤细胞株并分别命名为3C9,3F7,2G10.用ELISA方法对所获得的3个杂交瘤细胞株进行亚型鉴定均为IgG2a,kappa链.间接ELISA效价测定结果分别为3C9:1.024×10~7,3F7:2.56×10~6,2G10:1.28×10~6.此3株杂交瘤细胞所分泌的单克隆抗体均能与本研究室保存的其他3种不同的CGMMV分离物发生特异性反应,而不与其他3种同属成员病毒-烟草花叶病毒(Tobacco mosaic virus,TMV)、辣椒轻斑驳病毒(Pepper mild mottle virus,PMMoV)、齿瓣兰环斑病毒(Odontoglossum ringspot virus,ORSV)发生反应.%The Cucumber green mottle mosaic virus(CGMMV)was injected to BALB/c mice whose cells were fused with myeloma cells SP2/0 3 days after the final injection.On the selective medium,3 hybridoma cell lines were obtained by the finite dilution and indirect-ELISA.The 3 lines can secrete monoclonal antibody(McAbs)stably and were named as 3C9,3F7 and 2G10.respectively.The subtypes were identified by ELISA as IgG2a,Kappa light chain;the titration was examined,which were 3C9:1.024×10~7,3F7:2.56×10~6 and 2G10:1.28×10~6,respectively.By indirect-ELISA,3C9,3F7 and 2G10 were demonstrated to react specifically to the three different CGMMV isolates from other origins,whereas 3C9,3F7 and 2G10 did not response to either of the members of the same genus,TMV,PMMV or ORSV.