抗SARS冠状病毒S1蛋白N端249至667的单克隆抗体的制备与鉴定(英文)  

Preparation and characterization of monoclonal antibodies against S1domain at N-terminal residues 249 to 667 of SARS-associated coron-avirus S1 protein

作　　者：温坤[1] 梅亚波[1] 丘立文[1] 廖志勇[1] 袁国勇[2] 车小燕[1] 

机构地区：[1]第一军医大学珠江医院中心实验室,广东广州510282 [2]香港大学微生物学系

出　　处：《第一军医大学学报》2004年第1期1-6,共6页Journal of First Military Medical University

基　　金：supported as the Key Medical Research Project for SARS Prevention sponsored by the Ministry of Science and Technology of China and by Guangdong Province

摘　　要：目的在获得了具有免疫原性的SARS冠状病毒S1蛋白片段的基础上,制备和鉴定特异性抗该段S1蛋白单克隆抗体(mAb)。方法原核表达含S蛋白受体结合区的SARS冠状病毒S1蛋白片段S1c(N端249-667氨基酸残基),其免疫原性经SARS病人恢复期血清鉴定后免疫BALB/c小鼠,按常规方法制备单克隆抗体,并采用ELISA间接法、免疫荧光和免疫印迹进行筛选和鉴定。结果筛选出3株特异性针对SARS冠状病毒S1蛋白N端249-667的mAb杂交瘤细胞株,IgG亚类鉴定1株为IgG1,2株为IgG2a,经免疫荧光鉴定与人冠状病毒株229E和OC43无交叉反应。结论获得3株抗SARS冠状病毒S蛋白受体结合区特异性单克隆抗体,为建立新的SARS冠状病毒检测方法的和进一步研究S蛋白的功能奠定了基础。Objective To prepare and characterize monoclonal antibodies (mAbs) against S1 protein of severe acute respirato-ry syndrome (SARS)-associated coronavirus (SARS-CoV). Methods6-His-tagged recombinant fragment at N-terminal residues 249 to 667 of SARS-CoV S1 protein including S-protein receptor-binding domain was e xpressed in E.coli. The im-munogenicity of this S1 domain was identified and use d to immunize BALB/c mice for the production of hybridomas. The identification of the mAbs against this S1 domain was performed using indirect enzyme-linked im munosorbent assay (ELISA), indirect immunofluorescence assay (IFA) and Western blotting, respectively. Results Three hybridomas producing mAbs spe-cific to the S1 domain was obtained, with a relative molecular mass of 48 500. None of the 3 mAbs were reactive with human coronaviruses 229E and OC43. Two of the mAbs wer e IgG2a isotype, and the other was IgG1. Conclusion This is the first re-port of mAbs produced against S-protein receptor-binding domain of SARS-CoV. The 3 S1-s pecific mAbs may be useful for further study of the function of the S protein a nd for diagnosis of SARS-CoV infection.

关 键 词：SARS 严重急性呼吸综合症 传染性非典型肺炎 冠状病毒 S1蛋白 N端249-667 单克隆抗体 制备 鉴定 

分 类 号：R392[医药卫生—免疫学]