Identification of two antigenic epitopes on SARS-CoV spike protein.
Identification of two antigenic epitopes on SARS-CoV spike protein.
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DOI：
10.1016/j.bbrc.2004.05.066
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发表时间：
2004-07-02
影响因子：
3.1
通讯作者：
Tong G
中科院分区：
文献类型：
--
作者：
Hua R;Zhou Y;Wang Y;Hua Y;Tong G
S蛋白是SARS冠状病毒的主要结构蛋白。它在与受体相互作用和诱导中和抗体中起重要作用。本研究通过生物信息学分析，预测了SARS冠状病毒S蛋白的6个初步抗原表位（S1、S2、S3、S4、S5、S6），并合成了S1-S2-S3-S4-S5-S6多表位嵌合基因，将其与pGEX-6p-1下游GST基因融合。Western blotting结果表明SARS患者恢复期血清能够识别重组融合蛋白。针对该融合蛋白开发了许多单克隆抗体。进一步将6个预测表位基因分别与pGEX-6p-1的GST融合，并在大肠杆菌BL 21中表达。其中S5与抗SARS-CoV纤突蛋白单克隆抗体D3 C5反应，S2与抗SARS-CoV纤突蛋白单克隆抗体D3 D1反应。单克隆抗体D3 C5和D3 D1识别的抗原表位均为线性，分别对应SARS-CoV刺突蛋白的447-458和789-799个氨基酸。SARS-CoV S蛋白抗原表位的鉴定为SARS的预防、治疗和诊断提供了基础。
The spike (S) protein of severe acute respiratory syndrome-coronavirus (SARS-CoV) is a major virion structural protein. It plays an important role in interaction with receptor and inducing neutralizing antibodies. In the study, six tentative antigenic epitopes (S1 S2 S3 S4 S5 S6) of the spike protein of SARS-CoV were predicted by bio-informatics analysis, and a multi-epitope chimeric gene of S1–S2–S3–S4–S5–S6 was synthesized and fused to downstream GST gene in pGEX-6p-1. The Western blotting demonstrated that SARS patient convalescent serum could recognize the recombinant fusion protein. A number of monoclonal antibodies were developed against the fusion protein. In further, the six predicted epitope genes were individually fused to GST of pGEX-6p-1 and expressed in Escherichia coli BL21, respectively. Among six fusion peptides, S5 reacted with monoclonal antibody D3C5 and S2 reacted with monoclonal antibody D3D1 against spike protein of SARS-CoV. The epitopes recognized by monoclonal antibodies D3C5 and D3D1 are linear, and correspond to 447–458 and 789–799 amino acids of spike protein of SARS-CoV, respectively. Identification of antigenic epitope of spike protein of SARS-CoV could provide the basis for the development of immunity-based prophylactic, therapeutic, and diagnostic techniques for the control of severe acute respiratory syndrome.
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影响因子：
64.8
作者：
Li W;Moore MJ;Vasilieva N;Sui J;Wong SK;Berne MA;Somasundaran M;Sullivan JL;Luzuriaga K;Greenough TC;Choe H;Farzan M
通讯作者：
Farzan M
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：
--
作者：
Qin, E;Zhu, QY;Yang, HM
通讯作者：
Yang, HM
影响因子：
3.7
作者：
Gallagher TM;Buchmeier MJ
通讯作者：
Buchmeier MJ
DOI：
10.1016/j.bbrc.2003.11.054
发表时间：
2003-12-26
影响因子：
3.1
作者：
Xiao X;Chakraborti S;Dimitrov AS;Gramatikoff K;Dimitrov DS
通讯作者：
Dimitrov DS