Antigenicity and receptor-binding ability of recombinant SARS coronavirus spike protein.
Antigenicity and receptor-binding ability of recombinant SARS coronavirus spike protein.
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DOI：
10.1016/j.bbrc.2003.11.180
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发表时间：
2004-01-23
影响因子：
3.1
通讯作者：
Hsiang CY
中科院分区：
文献类型：
--
作者：
Ho TY;Wu SL;Cheng SE;Wei YC;Huang SP;Hsiang CY
严重急性呼吸系统综合征（SARS）是一种与新型冠状病毒相关的新兴传染病，在世界范围内引起暴发。SARS冠状病毒（SARS- cov）是一种包膜RNA病毒，它含有几种结构蛋白。在这些蛋白中，刺突蛋白(S)负责与特定的细胞受体结合，是主要的抗原决定因子，可诱导中和抗体。为了分析SARS-CoV S蛋白的抗原性和受体结合能力，我们利用pET表达载体在大肠杆菌中表达了SARS-CoV S蛋白。经异丙基-β-d-硫代半乳糖苷诱导后，S蛋白以可溶性形式表达，经镍亲和层析纯化至均匀性。S蛋白回收率为0.2 ~ 0.3 mg/100 ml细菌培养物。经酶联免疫吸附和免疫印迹检测，重组S蛋白可被SARS患者血清识别，表明重组S蛋白仍具有抗原性。通过生物素化ELISA和以生物素标记S蛋白为探针的Western blot，我们在Vero细胞中鉴定出130-kDa和140-kDa蛋白可能是导致SARS-CoV感染的细胞受体。综上所述，重组S蛋白具有抗原性和受体结合能力，可能成为进一步开发SARS疫苗和抗SARS治疗的良好候选蛋白。
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-β-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.
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DOI：
10.1016/s0140-6736(03)13077-2
发表时间：
2003-04-19
期刊：
Lancet (London, England)
影响因子：
--
作者：
Peiris JS;Lai ST;Poon LL;Guan Y;Yam LY;Lim W;Nicholls J;Yee WK;Yan WW;Cheung MT;Cheng VC;Chan KH;Tsang DN;Yung RW;Ng TK;Yuen KY;SARS study group
通讯作者：
SARS study group
影响因子：
3.5
作者：
Mechin, MC;DerVartanian, M;Martin, C
通讯作者：
Martin, C
影响因子：
3.7
作者：
COLLINS, AR;KNOBLER, RL;BUCHMEIER, MJ
通讯作者：
BUCHMEIER, MJ
影响因子：
3.7
作者：
OLESZAK, EL;PERLMAN, S;LEIBOWITZ, JL
通讯作者：
LEIBOWITZ, JL
影响因子：
4.1
作者：
Hsiang, CY;Ho, TY;Chang, TJ
通讯作者：
Chang, TJ