A Strategy for Searching Antigenic Regions in the SARS-CoV Spike Protein
Identifieur interne : 006107 ( Main/Exploration ); précédent : 006106; suivant : 006108A 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 LiuSource :
- Genomics, Proteomics & Bioinformatics [ 1672-0229 ] ; 2003.
Descripteurs français
- KwdFr :
- Antigènes viraux (immunologie), Chromatographie en phase liquide à haute performance, Clonage moléculaire, Fragments peptidiques (), Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (génétique), Glycoprotéines membranaires (immunologie), Glycoprotéines membranaires (métabolisme), Humains, Masse moléculaire, Protéines de l'enveloppe virale (génétique), Protéines de l'enveloppe virale (immunologie), Protéines de l'enveloppe virale (métabolisme), Protéines recombinantes (génétique), Protéines recombinantes (immunologie), Spectrométrie de masse, Test ELISA, Vecteurs génétiques, Virus du SRAS (génétique), Virus du SRAS (immunologie), Virus du SRAS (métabolisme), Électrophorèse sur gel de polyacrylamide.
- MESH :
- génétique : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines recombinantes, Virus du SRAS.
- immunologie : Antigènes viraux, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines recombinantes, Virus du SRAS.
- métabolisme : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Virus du SRAS.
- Chromatographie en phase liquide à haute performance, Clonage moléculaire, Fragments peptidiques, Glycoprotéine de spicule des coronavirus, Humains, Masse moléculaire, Spectrométrie de masse, Test ELISA, Vecteurs génétiques, Électrophorèse sur gel de polyacrylamide.
English descriptors
- KwdEn :
- Antigens, Viral (immunology), Chromatography, High Pressure Liquid, Cloning, Molecular, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Genetic Vectors, Humans, Mass Spectrometry, Membrane Glycoproteins (genetics), Membrane Glycoproteins (immunology), Membrane Glycoproteins (metabolism), Molecular Weight, Peptide Fragments (chemistry), Recombinant Proteins (genetics), Recombinant Proteins (immunology), SARS Virus (genetics), SARS Virus (immunology), SARS Virus (metabolism), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (genetics), Viral Envelope Proteins (immunology), Viral Envelope Proteins (metabolism).
- MESH :
- chemical , chemistry : Peptide Fragments.
- chemical , genetics : Membrane Glycoproteins, Recombinant Proteins, Viral Envelope Proteins.
- chemical , immunology : Antigens, Viral, Membrane Glycoproteins, Recombinant Proteins, Viral Envelope Proteins.
- chemical , metabolism : Membrane Glycoproteins, Viral Envelope Proteins.
- genetics : SARS Virus.
- immunology : SARS Virus.
- metabolism : SARS Virus.
- Chromatography, High Pressure Liquid, Cloning, Molecular, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Genetic Vectors, Humans, Mass Spectrometry, Molecular Weight, Spike Glycoprotein, Coronavirus.
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
Affiliations:
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Le document en format XML
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uniqKey="Liu J" first="Jinxiu" last="Liu">Jinxiu Liu</name></author><author><name sortKey="Lin, Liang" sort="Lin, Liang" uniqKey="Lin L" first="Liang" last="Lin">Liang Lin</name></author><author><name sortKey="Li, Shuting" sort="Li, Shuting" uniqKey="Li S" first="Shuting" last="Li">Shuting Li</name></author><author><name sortKey="Wang, Hao" sort="Wang, Hao" uniqKey="Wang H" first="Hao" last="Wang">Hao Wang</name></author><author><name sortKey="Xia, Ji" sort="Xia, Ji" uniqKey="Xia J" first="Ji" last="Xia">Ji Xia</name></author><author><name sortKey="Zhao, Zhe" sort="Zhao, Zhe" uniqKey="Zhao Z" first="Zhe" last="Zhao">Zhe Zhao</name></author><author><name sortKey="Wen, Jie" sort="Wen, Jie" uniqKey="Wen J" first="Jie" last="Wen">Jie Wen</name></author><author><name sortKey="Zhou, Cuiqi" sort="Zhou, Cuiqi" uniqKey="Zhou C" first="Cuiqi" last="Zhou">Cuiqi Zhou</name></author><author><name sortKey="Wang, Jingqiang" sort="Wang, Jingqiang" uniqKey="Wang J" first="Jingqiang" last="Wang">Jingqiang Wang</name></author><author><name sortKey="Yin, Jianning" sort="Yin, Jianning" uniqKey="Yin J" first="Jianning" last="Yin">Jianning Yin</name></author><author><name sortKey="Xu, Ningzhi" sort="Xu, Ningzhi" uniqKey="Xu N" first="Ningzhi" last="Xu">Ningzhi Xu</name></author><author><name sortKey="Liu, Siqi" sort="Liu, Siqi" uniqKey="Liu S" first="Siqi" last="Liu">Siqi Liu</name></author></analytic><series><title level="j">Genomics, Proteomics & Bioinformatics</title><idno type="ISSN">1672-0229</idno><idno type="eISSN">2210-3244</idno><imprint><date when="2003">2003</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antigens, Viral (immunology)</term><term>Chromatography, High Pressure Liquid</term><term>Cloning, Molecular</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Genetic Vectors</term><term>Humans</term><term>Mass Spectrometry</term><term>Membrane Glycoproteins (genetics)</term><term>Membrane Glycoproteins (immunology)</term><term>Membrane Glycoproteins (metabolism)</term><term>Molecular Weight</term><term>Peptide Fragments (chemistry)</term><term>Recombinant Proteins (genetics)</term><term>Recombinant Proteins (immunology)</term><term>SARS Virus (genetics)</term><term>SARS Virus (immunology)</term><term>SARS Virus (metabolism)</term><term>Spike Glycoprotein, Coronavirus</term><term>Viral Envelope Proteins (genetics)</term><term>Viral Envelope Proteins (immunology)</term><term>Viral Envelope Proteins (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Antigènes viraux (immunologie)</term><term>Chromatographie en phase liquide à haute performance</term><term>Clonage moléculaire</term><term>Fragments peptidiques ()</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires (génétique)</term><term>Glycoprotéines membranaires (immunologie)</term><term>Glycoprotéines membranaires (métabolisme)</term><term>Humains</term><term>Masse moléculaire</term><term>Protéines de l'enveloppe virale (génétique)</term><term>Protéines de l'enveloppe virale (immunologie)</term><term>Protéines de l'enveloppe virale (métabolisme)</term><term>Protéines recombinantes (génétique)</term><term>Protéines recombinantes (immunologie)</term><term>Spectrométrie de masse</term><term>Test ELISA</term><term>Vecteurs génétiques</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (immunologie)</term><term>Virus du SRAS (métabolisme)</term><term>Électrophorèse sur gel de polyacrylamide</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Peptide Fragments</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Membrane Glycoproteins</term><term>Recombinant Proteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antigens, Viral</term><term>Membrane Glycoproteins</term><term>Recombinant Proteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Protéines recombinantes</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Antigènes viraux</term><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Protéines recombinantes</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Chromatography, High Pressure Liquid</term><term>Cloning, Molecular</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Genetic Vectors</term><term>Humans</term><term>Mass Spectrometry</term><term>Molecular Weight</term><term>Spike Glycoprotein, Coronavirus</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Chromatographie en phase liquide à haute performance</term><term>Clonage moléculaire</term><term>Fragments peptidiques</term><term>Glycoprotéine de spicule des coronavirus</term><term>Humains</term><term>Masse moléculaire</term><term>Spectrométrie de masse</term><term>Test ELISA</term><term>Vecteurs génétiques</term><term>Électrophorèse sur gel de polyacrylamide</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><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></div></front><back><div1 type="bibliography"><listBibl><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Peiris, J S" uniqKey="Peiris J">J.S. Peiris</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ksiazek, T G" uniqKey="Ksiazek T">T.G. Ksiazek</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Rota, P A" uniqKey="Rota P">P.A. Rota</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Marra, M A" uniqKey="Marra M">M.A. Marra</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Qin, E D" uniqKey="Qin E">E.D. Qin</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Drosten, C" uniqKey="Drosten C">C. Drosten</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chan, H L" uniqKey="Chan H">H.L. Chan</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ruan, Y J" uniqKey="Ruan Y">Y.J. Ruan</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Haring, J" uniqKey="Haring J">J. Haring</name></author><author><name sortKey="Perlman, S" uniqKey="Perlman S">S. Perlman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sturman, L S" uniqKey="Sturman L">L.S. Sturman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Taguchi, F" uniqKey="Taguchi F">F. Taguchi</name></author><author><name sortKey="Shimazaki, Y K" uniqKey="Shimazaki Y">Y.K. Shimazaki</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Krueger, D K" uniqKey="Krueger D">D.K. Krueger</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gallagher, T M" uniqKey="Gallagher T">T.M. Gallagher</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Dea, S" uniqKey="Dea S">S. Dea</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cavanagh, D" uniqKey="Cavanagh D">D. Cavanagh</name></author><author><name sortKey="Davis, P J" uniqKey="Davis P">P.J. Davis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Koch, G" uniqKey="Koch G">G. Koch</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ignjatovic, J" uniqKey="Ignjatovic J">J. Ignjatovic</name></author><author><name sortKey="Galli, L" uniqKey="Galli L">L. Galli</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Schikora, B M" uniqKey="Schikora B">B.M. Schikora</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Parker, S E" uniqKey="Parker S">S.E. Parker</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kida, K" uniqKey="Kida K">K. Kida</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yoo, D" uniqKey="Yoo D">D. Yoo</name></author><author><name sortKey="Deregt, D" uniqKey="Deregt D">D. Deregt</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zhu, Q Y" uniqKey="Zhu Q">Q.Y. Zhu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sambrook, J" uniqKey="Sambrook J">J. Sambrook</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list></list><tree><noCountry><name sortKey="Li, Shuting" sort="Li, Shuting" uniqKey="Li S" first="Shuting" last="Li">Shuting Li</name><name sortKey="Lin, Liang" sort="Lin, Liang" uniqKey="Lin L" first="Liang" last="Lin">Liang Lin</name><name sortKey="Liu, Jinxiu" sort="Liu, Jinxiu" uniqKey="Liu J" first="Jinxiu" last="Liu">Jinxiu Liu</name><name sortKey="Liu, Siqi" sort="Liu, Siqi" uniqKey="Liu S" first="Siqi" last="Liu">Siqi Liu</name><name sortKey="Ren, Yan" sort="Ren, Yan" uniqKey="Ren Y" first="Yan" last="Ren">Yan Ren</name><name sortKey="Wang, Hao" sort="Wang, Hao" uniqKey="Wang H" first="Hao" last="Wang">Hao Wang</name><name sortKey="Wang, Jingqiang" sort="Wang, Jingqiang" uniqKey="Wang J" first="Jingqiang" last="Wang">Jingqiang Wang</name><name sortKey="Wen, Jie" sort="Wen, Jie" uniqKey="Wen J" first="Jie" last="Wen">Jie Wen</name><name sortKey="Xia, Ji" sort="Xia, Ji" uniqKey="Xia J" first="Ji" last="Xia">Ji Xia</name><name sortKey="Xu, Ningzhi" sort="Xu, Ningzhi" uniqKey="Xu N" first="Ningzhi" last="Xu">Ningzhi Xu</name><name sortKey="Yin, Jianning" sort="Yin, Jianning" uniqKey="Yin J" first="Jianning" last="Yin">Jianning Yin</name><name sortKey="Zhao, Zhe" sort="Zhao, Zhe" uniqKey="Zhao Z" first="Zhe" last="Zhao">Zhe Zhao</name><name sortKey="Zhou, Cuiqi" sort="Zhou, Cuiqi" uniqKey="Zhou C" first="Cuiqi" last="Zhou">Cuiqi Zhou</name><name sortKey="Zhou, Zhengfeng" sort="Zhou, Zhengfeng" uniqKey="Zhou Z" first="Zhengfeng" last="Zhou">Zhengfeng Zhou</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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