抗SARS病毒融合蛋白单克隆抗体的制备与初步鉴定  

Preparation and Identification of McAbs against Recombinant SARS-CoV S and N Fusion Protein

作　　者：郑纪山[1] 李越希[1] 唐祖明[2] 高健[1] 周宗安[1] 邓小昭[1] 

机构地区：[1]南京军区军事医学研究所,江苏南京210002 [2]东南大学分子与生物分子电子学实验室,江苏南京210096

出　　处：《药物生物技术》2006年第2期91-94,共4页Pharmaceutical Biotechnology

摘　　要：用基因工程技术表达的SARS-CoV S蛋白片段和N蛋白片段的融合蛋白作抗原免疫Balb/c小鼠,将免疫鼠脾细胞与Sp2/0细胞融合,经ELISA法筛选阳性克隆及测定效价,用免疫印迹法进行特异性鉴定。最终获得2株(1F11和3D2)能稳定分泌抗SARS-CoV融合蛋白单克隆抗体的杂交瘤细胞。其培养上清的ELISA效价分别为1∶1024和1∶512;腹水效价分别为:1×10-6,2×10-5。1F11株识别融合蛋白上的N抗原表位,3D2识别其上的S抗原表位。该单抗可望成为快速诊断SARS-CoV感染的特异性抗体。To obtain hybridoma cell lines secreting McAbs against SARS associated coronavirus （SARS- CoV） S protein and N protein, Balb/c mice were immunized with the recombinant SARS-CoV S and N fusion protein . Spleen cells from Balb/c mice were fused with Sp2/0 myeloma cells. The specific McAbs were screened by ELISA and identified by western blot . Two hybridoma cell lines （1Fll and 3D2） secreting specific McAbs to SARS-CoV were selected and determined to recognize different epitops on S and N protein fusion protein. The titers of the two clones were as the following： 1：1024, 1..512（supernant） ; 1×10^-6,2×10^-5（AF）. McAbs 1Fll recognize the epitope on N protein, whereas 3D2 recognize the epitope on S protein. This result suggested that specific McAbs could be used for developing diagnosis reagent.

关 键 词：SARS病毒 融合蛋白 单克隆抗体 

分 类 号：R392.11[医药卫生—免疫学]