南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (03): 290-293.

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人冠状病毒SARS-CoV、229E和OC43核衣壳(N)蛋白单克隆抗体的制备及3株人冠状病毒N蛋白的抗原相关性观察

丘立文； 王压娣； 廖志勇； 温坤； 车小燕；   

  1. 南方医科大学珠江医院中心实验室； 南方医科大学珠江医院中心实验室 广东广州510282； 广东广州510282；
  • 出版日期:2006-03-20 发布日期:2006-03-20
  • 基金资助:
    国家科技部防治非典型肺炎科技攻关项目;广东省防治非典型肺炎科技攻关项目(粤科社字:200380号文)~~

Antigenicity analysis of nucleocapsid proteins of 3 human coronaviruses SARS-CoV,229E and OC43 with their monoclonal antibodies

QIU Li-wen,WANG Ya-di,LIAO Zhi-yong,WEN Kun,CHE Xiao-yan Central Laboratory,Zhujiang Hospital,Southern Medical University,Guangzhou 510282,China   

  1. 南方医科大学珠江医院中心实验室； 南方医科大学珠江医院中心实验室 广东广州510282； 广东广州510282；
  • Online:2006-03-20 Published:2006-03-20

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摘要： 目的通过制备和鉴定人冠状病毒SARS-CoV、229E和OC43核衣壳(N)蛋白单克隆抗体(mAb),观察3株人冠状病毒的N蛋白抗原相关性。方法分别用基因重组SARS-CoV、229E和OC43N蛋白免疫BALB/c小鼠,制备mAb,并采用ELISA间接法、免疫印迹和免疫荧光进行筛选和鉴定,同时用mAb分析3株人冠状病毒N蛋白之间的免疫交叉反应。结果获得多株抗SARS-CoV、229E和OC43 N蛋白mAb杂交瘤细胞株,ELISA间接法、免疫印迹和免疫荧光结果均显示,所有的mAb仅与各自病毒的N蛋白特异性反应,而在3株人冠状病毒N蛋白抗原之间不产生免疫交叉反应。结论获得的特异性抗人冠状病毒N蛋白mAb为进一步研究3株人冠状病毒的抗原决定簇相关性奠定了基础。 更多还原

Abstract: Objective To prepare and characterize monoclonal antibodies(mAbs) against the recombinant nucleocapsid(N) protein of 3 human coronaviruses SARS-CoV,229E and OC43 and study the antigenic relationship between the 3 N proteins.Methods BALB/c mice were immunized with the recombinant N proteins of SARS-CoV,229E and OC43 to obtain the mAbs by means of hybridoma.Screening and identification of the mAbs were performed using indirect enzyme-linked immunosorbent assay(ELISA),Western blotting and indirect immunofluorescence assay.Cross-reactivity between the N proteins of the 3 coronaviruses was analyzed with the prepared mAbs.Results The mAbs against the recombinant N proteins of SARS-CoV,229E and OC43 were obtained,which reacted specifically with the corresponding viral N protein as shown by indirect ELISA,Western blotting and indirect immunofluorescence assay.No cross-reactivity was found between the 3 N proteins.Conclusion The prepared mAbs against the recombinant N proteins may provide valuable assistance in studying antigenic relationships of N proteins between the 3 human coronaviruses.

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