[Construction and identification of nanobody phage display library targeting Middle East respiratory syndrome coronavirus].
Identifieur interne : 000B13 ( Main/Exploration ); précédent : 000B12; suivant : 000B14[Construction and identification of nanobody phage display library targeting Middle East respiratory syndrome coronavirus].
Auteurs : Lei He [République populaire de Chine] ; Jiangfan Li [République populaire de Chine] ; Shuo Ren [République populaire de Chine] ; Shihui Sun [République populaire de Chine] ; Yan Guo [République populaire de Chine] ; Hongjie Qiu [République populaire de Chine] ; Yuanxiang Liao [République populaire de Chine] ; Kaiyuan Ji [République populaire de Chine] ; Ruiwen Fan [République populaire de Chine] ; Guangyu Zhao [Oman] ; Yusen Zhou [République populaire de Chine]Source :
- Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology [ 1007-8738 ] ; 2017.
Descripteurs français
- KwdFr :
- MESH :
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : Antibodies, Viral, Single-Domain Antibodies.
- chemical , immunology : Antibodies, Viral, Single-Domain Antibodies.
- immunology : Middle East Respiratory Syndrome Coronavirus.
- Animals, Camelids, New World, Peptide Library.
Abstract
Objective To construct a phage display library of specific nano-antibodies against the Middle East respiratory syndrome coronavirus (MERS-CoV) and apply it to the screening of neutralizing nano-antibodies. Methods MERS-CoV receptor-binding domain (RBD) recombinant protein was used to immunize alpaca. After the last immunization, peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood and total RNA was extracted. The VHH gene was amplified by PCR and used to construct recombinant phages. TG1 Escherichia coli was transformed by these recombinant phages. A phage display library of specific nano-antibodies against the MERS-CoV were obtained and used to screen and characterize the nano-antibodies. Results The volume of this library of nano-antibodies was 1.31×108 and its abundance rate was 5.65×1010/mL. The ratio of VHH insertion in the constructed library reached 96%. There was a rich diversity of nano-antibodies in this library. Nano-antibodies with neutralizing activity were identified and expressed from this library. Conclusion We successfully constructed a library of phages which could be effectively applied to the screening of nano-antibodies against MERS-CoV virus.
PubMed: 29382428
Affiliations:
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University, Jinzhong 030801, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Shanxi Agricultural University, Jinzhong 030801</wicri:regionArea><wicri:noRegion>Jinzhong 030801</wicri:noRegion></affiliation></author><author><name sortKey="Fan, Ruiwen" sort="Fan, Ruiwen" uniqKey="Fan R" first="Ruiwen" last="Fan">Ruiwen Fan</name><affiliation wicri:level="1"><nlm:affiliation>Shanxi Agricultural University, Jinzhong 030801, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Shanxi Agricultural University, Jinzhong 030801</wicri:regionArea><wicri:noRegion>Jinzhong 030801</wicri:noRegion></affiliation></author><author><name sortKey="Zhao, Guangyu" sort="Zhao, Guangyu" uniqKey="Zhao G" first="Guangyu" last="Zhao">Guangyu Zhao</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China. *Corresponding authors, E-mail: guangyu0525@163.com.</nlm:affiliation><country wicri:rule="url">Oman</country><wicri:regionArea>State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China. *Corresponding authors</wicri:regionArea><wicri:noRegion>China. *Corresponding authors</wicri:noRegion></affiliation></author><author><name sortKey="Zhou, Yusen" sort="Zhou, Yusen" uniqKey="Zhou Y" first="Yusen" last="Zhou">Yusen Zhou</name><affiliation wicri:level="1"><nlm:affiliation>Guangxi Medical University, Nanning 530021; State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China. *Corresponding authors, E-mail: yszhou@bmi.ac.cn.</nlm:affiliation><country wicri:rule="url">République populaire de Chine</country><wicri:regionArea>Guangxi Medical University, Nanning 530021; State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China. *Corresponding authors</wicri:regionArea><wicri:noRegion>China. *Corresponding authors</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2017">2017</date><idno type="RBID">pubmed:29382428</idno><idno type="pmid">29382428</idno><idno type="wicri:Area/PubMed/Corpus">000A09</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">000A09</idno><idno type="wicri:Area/PubMed/Curation">000A09</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">000A09</idno><idno type="wicri:Area/PubMed/Checkpoint">000A55</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">000A55</idno><idno type="wicri:Area/Ncbi/Merge">001D22</idno><idno type="wicri:Area/Ncbi/Curation">001D22</idno><idno type="wicri:Area/Ncbi/Checkpoint">001D22</idno><idno type="wicri:doubleKey">1007-8738:2017:He L:construction:and:identification</idno><idno type="wicri:Area/Main/Merge">000B16</idno><idno type="wicri:Area/Main/Curation">000B13</idno><idno type="wicri:Area/Main/Exploration">000B13</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">[Construction and identification of nanobody phage display library targeting Middle East respiratory syndrome coronavirus].</title><author><name sortKey="He, Lei" sort="He, Lei" uniqKey="He L" first="Lei" last="He">Lei He</name><affiliation wicri:level="1"><nlm:affiliation>Guangxi Medical University, Nanning 530021; State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Guangxi Medical University, Nanning 530021; State Key Laboratory of Pathogen and Biosecurity, Beijing 100071</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Li, Jiangfan" sort="Li, Jiangfan" uniqKey="Li J" first="Jiangfan" last="Li">Jiangfan Li</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Pathogen and Biosecurity, Beijing 100071</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Ren, Shuo" sort="Ren, Shuo" uniqKey="Ren S" first="Shuo" last="Ren">Shuo Ren</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Pathogen and Biosecurity, Beijing 100071</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Sun, Shihui" sort="Sun, Shihui" uniqKey="Sun S" first="Shihui" last="Sun">Shihui Sun</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Pathogen and Biosecurity, Beijing 100071</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Guo, Yan" sort="Guo, Yan" uniqKey="Guo Y" first="Yan" last="Guo">Yan Guo</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Pathogen and Biosecurity, Beijing 100071</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Qiu, Hongjie" sort="Qiu, Hongjie" uniqKey="Qiu H" first="Hongjie" last="Qiu">Hongjie Qiu</name><affiliation wicri:level="1"><nlm:affiliation>Rocket Force Center for Disease Prevention and Control, Beijing 100094, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Rocket Force Center for Disease Prevention and Control, Beijing 100094</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Liao, Yuanxiang" sort="Liao, Yuanxiang" uniqKey="Liao Y" first="Yuanxiang" last="Liao">Yuanxiang Liao</name><affiliation wicri:level="1"><nlm:affiliation>Rocket Force Center for Disease Prevention and Control, Beijing 100094, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Rocket Force Center for Disease Prevention and Control, Beijing 100094</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Ji, Kaiyuan" sort="Ji, Kaiyuan" uniqKey="Ji K" first="Kaiyuan" last="Ji">Kaiyuan Ji</name><affiliation wicri:level="1"><nlm:affiliation>Shanxi Agricultural University, Jinzhong 030801, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Shanxi Agricultural University, Jinzhong 030801</wicri:regionArea><wicri:noRegion>Jinzhong 030801</wicri:noRegion></affiliation></author><author><name sortKey="Fan, Ruiwen" sort="Fan, Ruiwen" uniqKey="Fan R" first="Ruiwen" last="Fan">Ruiwen Fan</name><affiliation wicri:level="1"><nlm:affiliation>Shanxi Agricultural University, Jinzhong 030801, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Shanxi Agricultural University, Jinzhong 030801</wicri:regionArea><wicri:noRegion>Jinzhong 030801</wicri:noRegion></affiliation></author><author><name sortKey="Zhao, Guangyu" sort="Zhao, Guangyu" uniqKey="Zhao G" first="Guangyu" last="Zhao">Guangyu Zhao</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China. *Corresponding authors, E-mail: guangyu0525@163.com.</nlm:affiliation><country wicri:rule="url">Oman</country><wicri:regionArea>State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China. *Corresponding authors</wicri:regionArea><wicri:noRegion>China. *Corresponding authors</wicri:noRegion></affiliation></author><author><name sortKey="Zhou, Yusen" sort="Zhou, Yusen" uniqKey="Zhou Y" first="Yusen" last="Zhou">Yusen Zhou</name><affiliation wicri:level="1"><nlm:affiliation>Guangxi Medical University, Nanning 530021; State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China. *Corresponding authors, E-mail: yszhou@bmi.ac.cn.</nlm:affiliation><country wicri:rule="url">République populaire de Chine</country><wicri:regionArea>Guangxi Medical University, Nanning 530021; State Key Laboratory of Pathogen and Biosecurity, Beijing 100071, China. *Corresponding authors</wicri:regionArea><wicri:noRegion>China. *Corresponding authors</wicri:noRegion></affiliation></author></analytic><series><title level="j">Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology</title><idno type="ISSN">1007-8738</idno><imprint><date when="2017" type="published">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Viral (genetics)</term><term>Antibodies, Viral (immunology)</term><term>Camelids, New World</term><term>Middle East Respiratory Syndrome Coronavirus (immunology)</term><term>Peptide Library</term><term>Single-Domain Antibodies (genetics)</term><term>Single-Domain Antibodies (immunology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Anticorps antiviraux (génétique)</term><term>Anticorps antiviraux (immunologie)</term><term>Anticorps à domaine unique (génétique)</term><term>Anticorps à domaine unique (immunologie)</term><term>Banque de peptides</term><term>Camélidés du Nouveau Monde</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Antibodies, Viral</term><term>Single-Domain Antibodies</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Viral</term><term>Single-Domain Antibodies</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Anticorps antiviraux</term><term>Anticorps à domaine unique</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps antiviraux</term><term>Anticorps à domaine unique</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Middle East Respiratory Syndrome Coronavirus</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Camelids, New World</term><term>Peptide Library</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Banque de peptides</term><term>Camélidés du Nouveau Monde</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Objective To construct a phage display library of specific nano-antibodies against the Middle East respiratory syndrome coronavirus (MERS-CoV) and apply it to the screening of neutralizing nano-antibodies. Methods MERS-CoV receptor-binding domain (RBD) recombinant protein was used to immunize alpaca. After the last immunization, peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood and total RNA was extracted. The VHH gene was amplified by PCR and used to construct recombinant phages. TG1 Escherichia coli was transformed by these recombinant phages. A phage display library of specific nano-antibodies against the MERS-CoV were obtained and used to screen and characterize the nano-antibodies. Results The volume of this library of nano-antibodies was 1.31×10<sup>8</sup> and its abundance rate was 5.65×10<sup>10</sup>/mL. The ratio of VHH insertion in the constructed library reached 96%. There was a rich diversity of nano-antibodies in this library. Nano-antibodies with neutralizing activity were identified and expressed from this library. Conclusion We successfully constructed a library of phages which could be effectively applied to the screening of nano-antibodies against MERS-CoV virus.</div></front></TEI><affiliations><list><country><li>Oman</li><li>République populaire de Chine</li></country><settlement><li>Pékin</li></settlement></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="He, Lei" sort="He, Lei" uniqKey="He L" first="Lei" last="He">Lei He</name></noRegion><name sortKey="Fan, Ruiwen" sort="Fan, Ruiwen" uniqKey="Fan R" first="Ruiwen" last="Fan">Ruiwen Fan</name><name sortKey="Guo, Yan" sort="Guo, Yan" uniqKey="Guo Y" first="Yan" last="Guo">Yan Guo</name><name sortKey="Ji, Kaiyuan" sort="Ji, Kaiyuan" uniqKey="Ji K" first="Kaiyuan" last="Ji">Kaiyuan Ji</name><name sortKey="Li, Jiangfan" sort="Li, Jiangfan" uniqKey="Li J" first="Jiangfan" last="Li">Jiangfan Li</name><name sortKey="Liao, Yuanxiang" sort="Liao, Yuanxiang" uniqKey="Liao Y" first="Yuanxiang" last="Liao">Yuanxiang Liao</name><name sortKey="Qiu, Hongjie" sort="Qiu, Hongjie" uniqKey="Qiu H" first="Hongjie" last="Qiu">Hongjie Qiu</name><name sortKey="Ren, Shuo" sort="Ren, Shuo" uniqKey="Ren S" first="Shuo" last="Ren">Shuo Ren</name><name sortKey="Sun, Shihui" sort="Sun, Shihui" uniqKey="Sun S" first="Shihui" last="Sun">Shihui Sun</name><name sortKey="Zhou, Yusen" sort="Zhou, Yusen" uniqKey="Zhou Y" first="Yusen" last="Zhou">Yusen Zhou</name></country><country name="Oman"><noRegion><name sortKey="Zhao, Guangyu" sort="Zhao, Guangyu" uniqKey="Zhao G" first="Guangyu" last="Zhao">Guangyu Zhao</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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