SARS冠状病毒N蛋白单克隆抗体的快速、高效制备的方法研究 |
| |
摘 要: |
|
关 键 词: | SARS 冠状病毒 严重急性呼吸道综合征 发病机制 诊断 蛋白组学 N蛋白单克隆抗体 制备 |
Rapid and efficient preparation of monoclonal antibodies against SARS-associated coronavirus nucleocapsid protein by immunizing mice. |
| |
Authors: | Xiao-yan Che Li-wen Qiu Yu-xian Pan Hua Xu Wei Hao Zhi-yong Liao Ya-bo Mei Li-ya Zhang Zhuo-yue Wan Guo-yong Yuan Zhen Huang |
| |
Institution: | Central Laboratory, Zhujiang Hospital, First Military Medical University, Guangzhou 510282, China. linche@pub.guangzhou.gd.cn |
| |
Abstract: | OBJECTIVE: To develop a rapid and efficient method for preparing monoclonal antibodies (mAb) against SARS-associated coronavirus (SARS-Cov) nucleocapsid (N) protein. METHODS: BALB/c mice were injected with the recombinant N protein of SARS-Cov into the foot-pads for the immunization, and the popliteal lymph nodes were isolated 15 d later for mAb-producing hybridomas, from which the mAbs against the N protein of SARS-Cov were screened. The identification of the mAb against the N protein of SARS-Cov was performed using indirect enzyme-linked immunosorbent assay (ELISA), indirect fluorescent-antibody assay (IFA), and Western immunoblotting. RESULTS: Four strains of hybridomas were obtained that produced the mAb specific to the N protein without detectable cross-reactivity with other pathogens. Of the 4 strains, 2 were identified as the immunoglobulin G1 (IgG1) isotype, 1 IgG2a, and the other IgG2b, with affinity constants (Ka) of 2 of the strains being 4.14 x 10(-9)M and 3.19 x 10(-9)M respectively. CONCLUSION: This is the first report on the preparation of mAb that is specific to the SARS-Cov, and the high-specificity and high-affinity mAb produced by the 4 strains of hybridomas provide a basis for further researches on the pathogenesis and early diagnosis of SARS. |
| |
Keywords: | |
|
|