[Construction and screening of SARS-CoV S protein-specific phage displayed antigen library].
Identifieur interne : 002719 ( Ncbi/Curation ); précédent : 002718; suivant : 002720[Construction and screening of SARS-CoV S protein-specific phage displayed antigen library].
Auteurs : Rui-Ping Wu [République populaire de Chine] ; Jia-Zi Meng ; Yu-Xian HeSource :
- Bing du xue bao = Chinese journal of virology [ 1000-8721 ] ; 2013.
Descripteurs français
- KwdFr :
- Anticorps antiviraux (immunologie), Bactériophages (génétique), Bactériophages (métabolisme), Banque de peptides, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (génétique), Glycoprotéines membranaires (immunologie), Humains, Protéines de l'enveloppe virale (génétique), Protéines de l'enveloppe virale (immunologie), Syndrome respiratoire aigu sévère (immunologie), Syndrome respiratoire aigu sévère (virologie), Virus du SRAS (génétique), Virus du SRAS (immunologie), Épitopes (génétique), Épitopes (immunologie).
- MESH :
- génétique : Bactériophages, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Virus du SRAS, Épitopes.
- immunologie : Anticorps antiviraux, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Syndrome respiratoire aigu sévère, Virus du SRAS, Épitopes.
- métabolisme : Bactériophages.
- virologie : Syndrome respiratoire aigu sévère.
- Banque de peptides, Glycoprotéine de spicule des coronavirus, Humains.
English descriptors
- KwdEn :
- Antibodies, Viral (immunology), Bacteriophages (genetics), Bacteriophages (metabolism), Epitopes (genetics), Epitopes (immunology), Humans, Membrane Glycoproteins (genetics), Membrane Glycoproteins (immunology), Peptide Library, SARS Virus (genetics), SARS Virus (immunology), Severe Acute Respiratory Syndrome (immunology), Severe Acute Respiratory Syndrome (virology), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (genetics), Viral Envelope Proteins (immunology).
- MESH :
- chemical , genetics : Epitopes, Membrane Glycoproteins, Viral Envelope Proteins.
- chemical , immunology : Antibodies, Viral, Epitopes, Membrane Glycoproteins, Viral Envelope Proteins.
- genetics : Bacteriophages, SARS Virus.
- immunology : SARS Virus, Severe Acute Respiratory Syndrome.
- metabolism : Bacteriophages.
- virology : Severe Acute Respiratory Syndrome.
- Humans, Peptide Library, Spike Glycoprotein, Coronavirus.
Abstract
The aim of this study is to construct a SARS-CoV S protein-specific phage displayed antigen library for the epitope characterization of anti-S monoclonal antibodies (mAbs). First, the full-length gene of SARS-S protein was PCR amplified, purified and then digested with DNase I to obtain DNA fragments in the size range of 50-500 bp. The resulting fragments were blunt-end ligated to the modified phage display vector pComb3XSS. The reactions were electrotransformed into XL1-Blue and infected with VCSM13 helper phage. The SARS-CoV S protein-specific phage displayed antigen library was biopanned and screened against two anti-S mAbs, S-M1 and S-M2. The results showed that we successfully constructed the phage displayed antigen library with a size of 5.7 x 10(6). After three-rounds of biopanning, 14 positive phage clones for S-M1 and 15 for S-M2 were respectively identified. Sequence analyses revealed the possible epitopes of two mAbs. Therefore, the S protein-specific phage displayed antigen library provides a crucial platform for the epitope characterization of anti-S antibodies and it is highly valuable for development of SARS vaccines and diagnostics.
PubMed: 23905471
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pubmed:23905471Le document en format XML
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He</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2013">2013</date><idno type="RBID">pubmed:23905471</idno><idno type="pmid">23905471</idno><idno type="wicri:Area/PubMed/Corpus">001158</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001158</idno><idno type="wicri:Area/PubMed/Curation">001158</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001158</idno><idno type="wicri:Area/PubMed/Checkpoint">001063</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001063</idno><idno type="wicri:Area/Ncbi/Merge">002719</idno><idno type="wicri:Area/Ncbi/Curation">002719</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">[Construction and screening of SARS-CoV S protein-specific phage displayed antigen library].</title><author><name sortKey="Wu, Rui Ping" sort="Wu, Rui Ping" uniqKey="Wu R" first="Rui-Ping" last="Wu">Rui-Ping Wu</name><affiliation wicri:level="1"><nlm:affiliation>Institute of Medical Virology, School of Laboratory Medicine, Wenzhou Medical College, Wenzhou 325035, China. wuruiping_q@126.com</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Institute of Medical Virology, School of Laboratory Medicine, Wenzhou Medical College, Wenzhou 325035</wicri:regionArea><wicri:noRegion>Wenzhou 325035</wicri:noRegion></affiliation></author><author><name sortKey="Meng, Jia Zi" sort="Meng, Jia Zi" uniqKey="Meng J" first="Jia-Zi" last="Meng">Jia-Zi Meng</name></author><author><name sortKey="He, Yu Xian" sort="He, Yu Xian" uniqKey="He Y" first="Yu-Xian" last="He">Yu-Xian He</name></author></analytic><series><title level="j">Bing du xue bao = Chinese journal of virology</title><idno type="ISSN">1000-8721</idno><imprint><date when="2013" type="published">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibodies, Viral (immunology)</term><term>Bacteriophages (genetics)</term><term>Bacteriophages (metabolism)</term><term>Epitopes (genetics)</term><term>Epitopes (immunology)</term><term>Humans</term><term>Membrane Glycoproteins (genetics)</term><term>Membrane Glycoproteins (immunology)</term><term>Peptide Library</term><term>SARS Virus (genetics)</term><term>SARS Virus (immunology)</term><term>Severe Acute Respiratory Syndrome (immunology)</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Spike Glycoprotein, Coronavirus</term><term>Viral Envelope Proteins (genetics)</term><term>Viral Envelope Proteins (immunology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Anticorps antiviraux (immunologie)</term><term>Bactériophages (génétique)</term><term>Bactériophages (métabolisme)</term><term>Banque de peptides</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires (génétique)</term><term>Glycoprotéines membranaires (immunologie)</term><term>Humains</term><term>Protéines de l'enveloppe virale (génétique)</term><term>Protéines de l'enveloppe virale (immunologie)</term><term>Syndrome respiratoire aigu sévère (immunologie)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (immunologie)</term><term>Épitopes (génétique)</term><term>Épitopes (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Epitopes</term><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Viral</term><term>Epitopes</term><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Bacteriophages</term><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Bactériophages</term><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Virus du SRAS</term><term>Épitopes</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps antiviraux</term><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Syndrome respiratoire aigu sévère</term><term>Virus du SRAS</term><term>Épitopes</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Bacteriophages</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Bactériophages</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Humans</term><term>Peptide Library</term><term>Spike Glycoprotein, Coronavirus</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Banque de peptides</term><term>Glycoprotéine de spicule des coronavirus</term><term>Humains</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"> The aim of this study is to construct a SARS-CoV S protein-specific phage displayed antigen library for the epitope characterization of anti-S monoclonal antibodies (mAbs). First, the full-length gene of SARS-S protein was PCR amplified, purified and then digested with DNase I to obtain DNA fragments in the size range of 50-500 bp. The resulting fragments were blunt-end ligated to the modified phage display vector pComb3XSS. The reactions were electrotransformed into XL1-Blue and infected with VCSM13 helper phage. The SARS-CoV S protein-specific phage displayed antigen library was biopanned and screened against two anti-S mAbs, S-M1 and S-M2. The results showed that we successfully constructed the phage displayed antigen library with a size of 5.7 x 10(6). After three-rounds of biopanning, 14 positive phage clones for S-M1 and 15 for S-M2 were respectively identified. Sequence analyses revealed the possible epitopes of two mAbs. Therefore, the S protein-specific phage displayed antigen library provides a crucial platform for the epitope characterization of anti-S antibodies and it is highly valuable for development of SARS vaccines and diagnostics.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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