Characterization of neutralizing monoclonal antibodies recognizing a 15-residues epitope on the spike protein HR2 region of severe acute respiratory syndrome coronavirus (SARS-CoV).
Identifieur interne : 004905 ( Main/Exploration ); précédent : 004904; suivant : 004906Characterization of neutralizing monoclonal antibodies recognizing a 15-residues epitope on the spike protein HR2 region of severe acute respiratory syndrome coronavirus (SARS-CoV).
Auteurs : Szu-Chia Lai [Taïwan] ; Pele Choi-Sing Chong ; Chia-Tsui Yeh ; Levent Shih-Jen Liu ; Jia-Tsrong Jan ; Hsiang-Yun Chi ; Hwan-Wun Liu ; Ann Chen ; Yeau-Ching WangSource :
- Journal of biomedical science [ 1021-7770 ] ; 2005.
Descripteurs français
- KwdFr :
- Amorces ADN, Animaux, Anticorps monoclonaux (), Anticorps monoclonaux (immunologie), Données de séquences moléculaires, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (), Glycoprotéines membranaires (immunologie), Protéines de l'enveloppe virale (), Protéines de l'enveloppe virale (immunologie), Souris, Souris de lignée BALB C, Séquence d'acides aminés, Séquence nucléotidique, Technique d'immunofluorescence indirecte, Technique de Western, Test ELISA, Test de retard de migration électrophorétique, Tests de neutralisation, Virus du SRAS (immunologie), Épitopes (), Épitopes (immunologie).
- MESH :
- immunologie : Anticorps monoclonaux, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Virus du SRAS, Épitopes.
- Amorces ADN, Animaux, Anticorps monoclonaux, Données de séquences moléculaires, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Souris, Souris de lignée BALB C, Séquence d'acides aminés, Séquence nucléotidique, Technique d'immunofluorescence indirecte, Technique de Western, Test ELISA, Test de retard de migration électrophorétique, Tests de neutralisation, Épitopes.
English descriptors
- KwdEn :
- Amino Acid Sequence, Animals, Antibodies, Monoclonal (chemistry), Antibodies, Monoclonal (immunology), Base Sequence, Blotting, Western, DNA Primers, Electrophoretic Mobility Shift Assay, Enzyme-Linked Immunosorbent Assay, Epitopes (chemistry), Epitopes (immunology), Fluorescent Antibody Technique, Indirect, Membrane Glycoproteins (chemistry), Membrane Glycoproteins (immunology), Mice, Mice, Inbred BALB C, Molecular Sequence Data, Neutralization Tests, SARS Virus (immunology), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (chemistry), Viral Envelope Proteins (immunology).
- MESH :
- chemical , chemistry : Antibodies, Monoclonal, Epitopes, Membrane Glycoproteins, Viral Envelope Proteins.
- chemical , immunology : Antibodies, Monoclonal, Epitopes, Membrane Glycoproteins, Viral Envelope Proteins.
- immunology : SARS Virus.
- Amino Acid Sequence, Animals, Base Sequence, Blotting, Western, DNA Primers, Electrophoretic Mobility Shift Assay, Enzyme-Linked Immunosorbent Assay, Fluorescent Antibody Technique, Indirect, Mice, Mice, Inbred BALB C, Molecular Sequence Data, Neutralization Tests, Spike Glycoprotein, Coronavirus.
Abstract
The spike (S) glycoprotein is thought to play a complex and central role in the biology and pathogenesis of SARS coronavirus infection. In this study, a recombinant protein (rS268, corresponding to residues 268-1255 of SARS-CoV S protein) was expressed in Escherichia coli and was purified to near homogeneity. After immunization with rS268, S protein-specific BALB/c antisera and mAbs were induced and confirmed using ELISA, Western blot and IFA. Several BALB/c mAbs were found to be effectively to neutralize the infection of Vero E6 cells by SARS-CoV in a dose-dependent manner. Systematic epitope mapping showed that all these neutralizing mAbs recognized a 15-residues peptide (CB-119) corresponding to residues 1143-1157 (SPDVDLGDISGINAS) that was located to the second heptad repeat (HR2) region of the SARS-CoV spike protein. The peptide CB-119 could specifically inhibit the interaction of neutralizing mAbs and spike protein in a dose-dependent manner. Further, neutralizing mAbs, but not control mAbs, could specifically interact with CB-119 in a dose-dependent manner. Results implicated that the second heptad repeat region of spike protein could be a good target for vaccine development against SARS-CoV.
DOI: 10.1007/s11373-005-9004-3
PubMed: 16132115
Affiliations:
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Le document en format XML
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<term>Antibodies, Monoclonal (immunology)</term>
<term>Base Sequence</term>
<term>Blotting, Western</term>
<term>DNA Primers</term>
<term>Electrophoretic Mobility Shift Assay</term>
<term>Enzyme-Linked Immunosorbent Assay</term>
<term>Epitopes (chemistry)</term>
<term>Epitopes (immunology)</term>
<term>Fluorescent Antibody Technique, Indirect</term>
<term>Membrane Glycoproteins (chemistry)</term>
<term>Membrane Glycoproteins (immunology)</term>
<term>Mice</term>
<term>Mice, Inbred BALB C</term>
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<term>SARS Virus (immunology)</term>
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<term>Viral Envelope Proteins (chemistry)</term>
<term>Viral Envelope Proteins (immunology)</term>
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<term>Animaux</term>
<term>Anticorps monoclonaux ()</term>
<term>Anticorps monoclonaux (immunologie)</term>
<term>Données de séquences moléculaires</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Glycoprotéines membranaires ()</term>
<term>Glycoprotéines membranaires (immunologie)</term>
<term>Protéines de l'enveloppe virale ()</term>
<term>Protéines de l'enveloppe virale (immunologie)</term>
<term>Souris</term>
<term>Souris de lignée BALB C</term>
<term>Séquence d'acides aminés</term>
<term>Séquence nucléotidique</term>
<term>Technique d'immunofluorescence indirecte</term>
<term>Technique de Western</term>
<term>Test ELISA</term>
<term>Test de retard de migration électrophorétique</term>
<term>Tests de neutralisation</term>
<term>Virus du SRAS (immunologie)</term>
<term>Épitopes ()</term>
<term>Épitopes (immunologie)</term>
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<term>Epitopes</term>
<term>Membrane Glycoproteins</term>
<term>Viral Envelope Proteins</term>
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<term>Membrane Glycoproteins</term>
<term>Viral Envelope Proteins</term>
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<term>Glycoprotéines membranaires</term>
<term>Protéines de l'enveloppe virale</term>
<term>Virus du SRAS</term>
<term>Épitopes</term>
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<term>Animals</term>
<term>Base Sequence</term>
<term>Blotting, Western</term>
<term>DNA Primers</term>
<term>Electrophoretic Mobility Shift Assay</term>
<term>Enzyme-Linked Immunosorbent Assay</term>
<term>Fluorescent Antibody Technique, Indirect</term>
<term>Mice</term>
<term>Mice, Inbred BALB C</term>
<term>Molecular Sequence Data</term>
<term>Neutralization Tests</term>
<term>Spike Glycoprotein, Coronavirus</term>
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<term>Anticorps monoclonaux</term>
<term>Données de séquences moléculaires</term>
<term>Glycoprotéine de spicule des coronavirus</term>
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<front><div type="abstract" xml:lang="en">The spike (S) glycoprotein is thought to play a complex and central role in the biology and pathogenesis of SARS coronavirus infection. In this study, a recombinant protein (rS268, corresponding to residues 268-1255 of SARS-CoV S protein) was expressed in Escherichia coli and was purified to near homogeneity. After immunization with rS268, S protein-specific BALB/c antisera and mAbs were induced and confirmed using ELISA, Western blot and IFA. Several BALB/c mAbs were found to be effectively to neutralize the infection of Vero E6 cells by SARS-CoV in a dose-dependent manner. Systematic epitope mapping showed that all these neutralizing mAbs recognized a 15-residues peptide (CB-119) corresponding to residues 1143-1157 (SPDVDLGDISGINAS) that was located to the second heptad repeat (HR2) region of the SARS-CoV spike protein. The peptide CB-119 could specifically inhibit the interaction of neutralizing mAbs and spike protein in a dose-dependent manner. Further, neutralizing mAbs, but not control mAbs, could specifically interact with CB-119 in a dose-dependent manner. Results implicated that the second heptad repeat region of spike protein could be a good target for vaccine development against SARS-CoV.</div>
</front>
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<affiliations><list><country><li>Taïwan</li>
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<tree><noCountry><name sortKey="Chen, Ann" sort="Chen, Ann" uniqKey="Chen A" first="Ann" last="Chen">Ann Chen</name>
<name sortKey="Chi, Hsiang Yun" sort="Chi, Hsiang Yun" uniqKey="Chi H" first="Hsiang-Yun" last="Chi">Hsiang-Yun Chi</name>
<name sortKey="Chong, Pele Choi Sing" sort="Chong, Pele Choi Sing" uniqKey="Chong P" first="Pele Choi-Sing" last="Chong">Pele Choi-Sing Chong</name>
<name sortKey="Jan, Jia Tsrong" sort="Jan, Jia Tsrong" uniqKey="Jan J" first="Jia-Tsrong" last="Jan">Jia-Tsrong Jan</name>
<name sortKey="Liu, Hwan Wun" sort="Liu, Hwan Wun" uniqKey="Liu H" first="Hwan-Wun" last="Liu">Hwan-Wun Liu</name>
<name sortKey="Liu, Levent Shih Jen" sort="Liu, Levent Shih Jen" uniqKey="Liu L" first="Levent Shih-Jen" last="Liu">Levent Shih-Jen Liu</name>
<name sortKey="Wang, Yeau Ching" sort="Wang, Yeau Ching" uniqKey="Wang Y" first="Yeau-Ching" last="Wang">Yeau-Ching Wang</name>
<name sortKey="Yeh, Chia Tsui" sort="Yeh, Chia Tsui" uniqKey="Yeh C" first="Chia-Tsui" last="Yeh">Chia-Tsui Yeh</name>
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<country name="Taïwan"><noRegion><name sortKey="Lai, Szu Chia" sort="Lai, Szu Chia" uniqKey="Lai S" first="Szu-Chia" last="Lai">Szu-Chia Lai</name>
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