Antigenicity and receptor-binding ability of recombinant SARS coronavirus spike protein

被引:31
作者
Ho, TY
Wu, SL
Cheng, SE
Wei, YC
Huang, SP
Hsiang, CY [1 ]
机构
[1] China Med Univ, Dept Microbiol, Taichung, Taiwan
[2] China Med Univ, Inst Chinese Med Sci, Taichung, Taiwan
[3] China Med Univ, Dept Biochem, Taichung, Taiwan
关键词
SARS; coronavirus; spike; expression; antigenicity; receptor-binding;
D O I
10.1016/j.bbrc.2003.11.180
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Severe acute respiratory syndrome (SARS) is an emerging infectious disease associated with a novel coronavirus and causing worldwide outbreaks. SARS coronavirus (SARS-CoV) is an enveloped RNA virus, which contains several structural proteins. Among these proteins, spike (S) protein is responsible for binding to specific cellular receptors and is a major antigenic determinant, which induces neutralizing antibody. In order to analyze the antigenicity and receptor-binding ability of SARS-CoV S protein, we expressed the S protein in Escherichia coli using a pET expression vector. After the isopropyl-p-D-thiogalactoside induction, S protein was expressed in the soluble form and purified by nickel-affinity chromatography to homogeneity. The amount of S protein recovered was 0.2-0.3 mg/100 ml bacterial culture. The S protein was recognized by sera from SARS patients by ELISA and Western blot, which indicated that recombinant S protein retained its antigenicity. By biotinylated ELISA and Western blot using biotin-labeled S protein as the probe, we identified 130-kDa and 140-kDa proteins in Vero cells that might be the cellular receptors responsible for SARS-CoV infection. Taken together, these results suggested that recombinant S protein exhibited the antigenicity and receptor-binding ability, and it could be a good candidate for further developing SARS vaccine and anti-SARS therapy. (C) 2003 Elsevier Inc. All rights reserved.
引用
收藏
页码:938 / 947
页数:10
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