Qiu Li-wen, Wang Ya-di, Liao Zhi-yong, Wen Kun, Che Xiao-yan
Central Laboratory, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2006 Mar;26(3):290-3.
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.
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.
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.
The prepared mAbs against the recombinant N proteins may provide valuable assistance in studying antigenic relationships of N proteins between the 3 human coronaviruses.
制备并鉴定针对三种人类冠状病毒SARS-CoV、229E和OC43重组核衣壳（N）蛋白的单克隆抗体（mAb），并研究这三种N蛋白之间的抗原关系。
用SARS-CoV、229E和OC43的重组N蛋白免疫BALB/c小鼠，通过杂交瘤技术获得单克隆抗体。使用间接酶联免疫吸附测定（ELISA）、蛋白质印迹法和间接免疫荧光测定法对单克隆抗体进行筛选和鉴定。用制备的单克隆抗体分析三种冠状病毒N蛋白之间的交叉反应性。
获得了针对SARS-CoV、229E和OC43重组N蛋白的单克隆抗体，间接ELISA、蛋白质印迹法和间接免疫荧光测定法显示这些抗体与相应的病毒N蛋白发生特异性反应。三种N蛋白之间未发现交叉反应性。
制备的针对重组N蛋白的单克隆抗体可能为研究三种人类冠状病毒N蛋白之间的抗原关系提供有价值的帮助。
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