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A Strategy for Searching Antigenic Regions in the SARS-CoV Spike Protein

Identifieur interne : 001026 ( Pmc/Curation ); précédent : 001025; suivant : 001027

A Strategy for Searching Antigenic Regions in the SARS-CoV Spike Protein

Auteurs : Yan Ren ; Zhengfeng Zhou ; Jinxiu Liu ; Liang Lin ; Shuting Li ; Hao Wang ; Ji Xia ; Zhe Zhao ; Jie Wen ; Cuiqi Zhou ; Jingqiang Wang ; Jianning Yin ; Ningzhi Xu ; Siqi Liu

Source :

RBID : PMC:5172407

Abstract

In the face of the worldwide threat of severe acute respiratory syndrome (SARS) to human life, some of the most urgent challenges are to develop fast and accurate analytical methods for early diagnosis of this disease as well as to create a safe anti-viral vaccine for prevention. To these ends, we investigated the antigenicity of the spike protein (S protein), a major structural protein in the SARS-coronavirus (SARS-CoV). Based upon the theoretical analysis for hydrophobicity of the S protein, 18 peptides were synthesized. Using Enzyme-Linked Immunosorbent Assay (ELISA), these peptides were screened in the sera from SARS patients. According to these results, two fragments of the S gene were amplified by PCR and cloned into pET-32a. Both S fragments were expressed in the BL-21 strain and further purified with an affinity chromatography. These recombinant S fragments were confirmed to have positive cross-reactions with SARS sera, either by Western blot or by ELISA. Our results demonstrated that the potential epitope regions were located at Codons 469–882 in the S protein, and one epitope site was located at Codons 599–620. Identification of antigenic regions in the SARS-CoV S protein may be important for the functional studies of this virus or the development of clinical diagnosis.


Url:
DOI: 10.1016/S1672-0229(03)01026-X
PubMed: 15629033
PubMed Central: 5172407

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<p>In the face of the worldwide threat of severe acute respiratory syndrome (SARS) to human life, some of the most urgent challenges are to develop fast and accurate analytical methods for early diagnosis of this disease as well as to create a safe anti-viral vaccine for prevention. To these ends, we investigated the antigenicity of the spike protein (S protein), a major structural protein in the SARS-coronavirus (SARS-CoV). Based upon the theoretical analysis for hydrophobicity of the S protein, 18 peptides were synthesized. Using Enzyme-Linked Immunosorbent Assay (ELISA), these peptides were screened in the sera from SARS patients. According to these results, two fragments of the S gene were amplified by PCR and cloned into pET-32a. Both S fragments were expressed in the BL-21 strain and further purified with an affinity chromatography. These recombinant S fragments were confirmed to have positive cross-reactions with SARS sera, either by Western blot or by ELISA. Our results demonstrated that the potential epitope regions were located at Codons 469–882 in the S protein, and one epitope site was located at Codons 599–620. Identification of antigenic regions in the SARS-CoV S protein may be important for the functional studies of this virus or the development of clinical diagnosis.</p>
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<article-title>A Strategy for Searching Antigenic Regions in the SARS-CoV Spike Protein</article-title>
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<contrib-group>
<contrib contrib-type="author" id="au0005">
<name>
<surname>Ren</surname>
<given-names>Yan</given-names>
</name>
<xref rid="fn1" ref-type="fn">*</xref>
</contrib>
<contrib contrib-type="author" id="au0010">
<name>
<surname>Zhou</surname>
<given-names>Zhengfeng</given-names>
</name>
<xref rid="fn1" ref-type="fn">*</xref>
</contrib>
<contrib contrib-type="author" id="au0015">
<name>
<surname>Liu</surname>
<given-names>Jinxiu</given-names>
</name>
<xref rid="fn1" ref-type="fn">*</xref>
</contrib>
<contrib contrib-type="author" id="au0020">
<name>
<surname>Lin</surname>
<given-names>Liang</given-names>
</name>
</contrib>
<contrib contrib-type="author" id="au0025">
<name>
<surname>Li</surname>
<given-names>Shuting</given-names>
</name>
</contrib>
<contrib contrib-type="author" id="au0030">
<name>
<surname>Wang</surname>
<given-names>Hao</given-names>
</name>
</contrib>
<contrib contrib-type="author" id="au0035">
<name>
<surname>Xia</surname>
<given-names>Ji</given-names>
</name>
</contrib>
<contrib contrib-type="author" id="au0040">
<name>
<surname>Zhao</surname>
<given-names>Zhe</given-names>
</name>
</contrib>
<contrib contrib-type="author" id="au0045">
<name>
<surname>Wen</surname>
<given-names>Jie</given-names>
</name>
</contrib>
<contrib contrib-type="author" id="au0050">
<name>
<surname>Zhou</surname>
<given-names>Cuiqi</given-names>
</name>
</contrib>
<contrib contrib-type="author" id="au0055">
<name>
<surname>Wang</surname>
<given-names>Jingqiang</given-names>
</name>
</contrib>
<contrib contrib-type="author" id="au0060">
<name>
<surname>Yin</surname>
<given-names>Jianning</given-names>
</name>
</contrib>
<contrib contrib-type="author" id="au0065">
<name>
<surname>Xu</surname>
<given-names>Ningzhi</given-names>
</name>
</contrib>
<contrib contrib-type="author" id="au0070">
<name>
<surname>Liu</surname>
<given-names>Siqi</given-names>
</name>
<email>siqiliu@genomics.org.cn</email>
<xref rid="cor1" ref-type="corresp">#</xref>
</contrib>
</contrib-group>
<aff id="aff0005">Beijing Genomics Institute, Chinese Academy of Sciences, Beijing 101300, China</aff>
<aff id="aff0010">Beijing Proteomics Institute, Beijing 101300, China</aff>
<author-notes>
<corresp id="cor1">
<label>#</label>
Corresponding author.
<email>siqiliu@genomics.org.cn</email>
</corresp>
<fn id="fn1">
<label>*</label>
<p id="ntp0005">These authors contributed equally to this work.</p>
</fn>
</author-notes>
<pub-date pub-type="pmc-release">
<day>28</day>
<month>11</month>
<year>2016</year>
</pub-date>
<pmc-comment> PMC Release delay is 0 months and 0 days and was based on .</pmc-comment>
<pub-date pub-type="ppub">
<month>8</month>
<year>2003</year>
</pub-date>
<pub-date pub-type="epub">
<day>28</day>
<month>11</month>
<year>2016</year>
</pub-date>
<volume>1</volume>
<issue>3</issue>
<fpage>207</fpage>
<lpage>215</lpage>
<permissions>
<copyright-statement>Copyright © 2003 Beijing Institute of Genomics, the Chinese Academy of Sciences and the Genetics Society of China. Production and hosting by Elsevier B.V.</copyright-statement>
<copyright-year>2003</copyright-year>
<copyright-holder>Beijing Institute of Genomics, the Chinese Academy of Sciences and the Genetics Society of China</copyright-holder>
<license>
<license-p>Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.</license-p>
</license>
</permissions>
<abstract id="ab0005">
<p>In the face of the worldwide threat of severe acute respiratory syndrome (SARS) to human life, some of the most urgent challenges are to develop fast and accurate analytical methods for early diagnosis of this disease as well as to create a safe anti-viral vaccine for prevention. To these ends, we investigated the antigenicity of the spike protein (S protein), a major structural protein in the SARS-coronavirus (SARS-CoV). Based upon the theoretical analysis for hydrophobicity of the S protein, 18 peptides were synthesized. Using Enzyme-Linked Immunosorbent Assay (ELISA), these peptides were screened in the sera from SARS patients. According to these results, two fragments of the S gene were amplified by PCR and cloned into pET-32a. Both S fragments were expressed in the BL-21 strain and further purified with an affinity chromatography. These recombinant S fragments were confirmed to have positive cross-reactions with SARS sera, either by Western blot or by ELISA. Our results demonstrated that the potential epitope regions were located at Codons 469–882 in the S protein, and one epitope site was located at Codons 599–620. Identification of antigenic regions in the SARS-CoV S protein may be important for the functional studies of this virus or the development of clinical diagnosis.</p>
</abstract>
<kwd-group id="keys0005">
<title>Key words</title>
<kwd>SARS</kwd>
<kwd>the spike protein</kwd>
<kwd>expression</kwd>
<kwd>antigenicity</kwd>
</kwd-group>
</article-meta>
</front>
</pmc>
</record>

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