Construction of a single chain variable fragment antibody (scFv) against tetrodotoxin (TTX) and its interaction with TTX.
Identifieur interne : 001B50 ( Main/Exploration ); précédent : 001B49; suivant : 001B51Construction of a single chain variable fragment antibody (scFv) against tetrodotoxin (TTX) and its interaction with TTX.
Auteurs : Rongzhi Wang [République populaire de Chine] ; Ailing Huang [République populaire de Chine] ; Licai Liu [République populaire de Chine] ; Shuangshuang Xiang [République populaire de Chine] ; Xiufeng Li [République populaire de Chine] ; Sumei Ling [République populaire de Chine] ; Lei Wang [République populaire de Chine] ; Tun Lu [République populaire de Chine] ; Shihua Wang [République populaire de Chine]Source :
- Toxicon : official journal of the International Society on Toxinology [ 1879-3150 ] ; 2014.
Descripteurs français
- KwdFr :
- Affinité des anticorps (MeSH), Analyse de séquence de protéine (MeSH), Anticorps neutralisants (composition chimique), Anticorps à chaîne unique (composition chimique), Banque de peptides (MeSH), Données de séquences moléculaires (MeSH), Modèles moléculaires (MeSH), Mutation ponctuelle (MeSH), Sites de fixation (MeSH), Séquence d'acides aminés (MeSH), Séquence nucléotidique (MeSH), Tétrodotoxine (immunologie).
- MESH :
- composition chimique : Anticorps neutralisants, Anticorps à chaîne unique.
- immunologie : Tétrodotoxine.
- Affinité des anticorps, Analyse de séquence de protéine, Banque de peptides, Données de séquences moléculaires, Modèles moléculaires, Mutation ponctuelle, Sites de fixation, Séquence d'acides aminés, Séquence nucléotidique.
English descriptors
- KwdEn :
- Amino Acid Sequence (MeSH), Antibodies, Neutralizing (chemistry), Antibody Affinity (MeSH), Base Sequence (MeSH), Binding Sites (MeSH), Models, Molecular (MeSH), Molecular Sequence Data (MeSH), Peptide Library (MeSH), Point Mutation (MeSH), Sequence Analysis, Protein (MeSH), Single-Chain Antibodies (chemistry), Tetrodotoxin (immunology).
- MESH :
- chemical , chemistry : Antibodies, Neutralizing, Single-Chain Antibodies.
- chemical , immunology : Tetrodotoxin.
- Amino Acid Sequence, Antibody Affinity, Base Sequence, Binding Sites, Models, Molecular, Molecular Sequence Data, Peptide Library, Point Mutation, Sequence Analysis, Protein.
Abstract
Tetrodotoxin (TTX) is a small molecular weight neurotoxin that occludes voltage-gated sodium channels in nerve and muscle tissue, resulting in respiratory paralysis and death. A high affinity antibody that can neutralize the toxicity of TTX is still lacking, so it is very important to prepare an antibody for TTX therapy and detection. In the present study, a chemical method was used to prepare the tetrodotoxin complete antigen, and a small amount, repeatedly immunity way was carried to immunize 4 mice. The amplified genes encoding monoclonal antibodies against TTX were used to construct the phage display library. After six rounds of biopanning, an antibody named scFv-T53 was characterized from clones showing high affinity and specific to TTX, and its affinity constant was 1.1 × 10(6) L/mol. Three dimensional structure of the scFv-T53 was constructed by computer modeling, and TTX was docked to the scFv-T53 model to obtain the structure of the binding complex. Two predicted essential amino acids, K183 and I189, were mutated to verify the theoretical model. Both mutants lost binding activity significantly against TTX as predicted by the theoretical model. Hence, the above results will be useful for screening the high affinity anti-TTX scFv mutants.
DOI: 10.1016/j.toxicon.2014.02.021
PubMed: 24613546
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Construction of a single chain variable fragment antibody (scFv) against tetrodotoxin (TTX) and its interaction with TTX.</title><author><name sortKey="Wang, Rongzhi" sort="Wang, Rongzhi" uniqKey="Wang R" first="Rongzhi" last="Wang">Rongzhi Wang</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Huang, Ailing" sort="Huang, Ailing" uniqKey="Huang A" first="Ailing" last="Huang">Ailing Huang</name><affiliation wicri:level="1"><nlm:affiliation>Bioengineering and Bioscience Department, Fuzhou University, Fuzhou 350108, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Bioengineering and Bioscience Department, Fuzhou University, Fuzhou 350108</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Liu, Licai" sort="Liu, Licai" uniqKey="Liu L" first="Licai" last="Liu">Licai Liu</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Xiang, Shuangshuang" sort="Xiang, Shuangshuang" uniqKey="Xiang S" first="Shuangshuang" last="Xiang">Shuangshuang Xiang</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Li, Xiufeng" sort="Li, Xiufeng" uniqKey="Li X" first="Xiufeng" last="Li">Xiufeng Li</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Ling, Sumei" sort="Ling, Sumei" uniqKey="Ling S" first="Sumei" last="Ling">Sumei Ling</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Wang, Lei" sort="Wang, Lei" uniqKey="Wang L" first="Lei" last="Wang">Lei Wang</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Lu, Tun" sort="Lu, Tun" uniqKey="Lu T" first="Tun" last="Lu">Tun Lu</name><affiliation wicri:level="1"><nlm:affiliation>Bioengineering and Bioscience Department, Fuzhou University, Fuzhou 350108, China. Electronic address: lvtun@yahoo.com.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Bioengineering and Bioscience Department, Fuzhou University, Fuzhou 350108</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Wang, Shihua" sort="Wang, Shihua" uniqKey="Wang S" first="Shihua" last="Wang">Shihua Wang</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China. Electronic address: wshyyl@sina.com.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2014">2014</date><idno type="RBID">pubmed:24613546</idno><idno type="pmid">24613546</idno><idno type="doi">10.1016/j.toxicon.2014.02.021</idno><idno type="wicri:Area/Main/Corpus">001A84</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">001A84</idno><idno type="wicri:Area/Main/Curation">001A84</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">001A84</idno><idno type="wicri:Area/Main/Exploration">001A84</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Construction of a single chain variable fragment antibody (scFv) against tetrodotoxin (TTX) and its interaction with TTX.</title><author><name sortKey="Wang, Rongzhi" sort="Wang, Rongzhi" uniqKey="Wang R" first="Rongzhi" last="Wang">Rongzhi Wang</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Huang, Ailing" sort="Huang, Ailing" uniqKey="Huang A" first="Ailing" last="Huang">Ailing Huang</name><affiliation wicri:level="1"><nlm:affiliation>Bioengineering and Bioscience Department, Fuzhou University, Fuzhou 350108, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Bioengineering and Bioscience Department, Fuzhou University, Fuzhou 350108</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Liu, Licai" sort="Liu, Licai" uniqKey="Liu L" first="Licai" last="Liu">Licai Liu</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Xiang, Shuangshuang" sort="Xiang, Shuangshuang" uniqKey="Xiang S" first="Shuangshuang" last="Xiang">Shuangshuang Xiang</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Li, Xiufeng" sort="Li, Xiufeng" uniqKey="Li X" first="Xiufeng" last="Li">Xiufeng Li</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Ling, Sumei" sort="Ling, Sumei" uniqKey="Ling S" first="Sumei" last="Ling">Sumei Ling</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Wang, Lei" sort="Wang, Lei" uniqKey="Wang L" first="Lei" last="Wang">Lei Wang</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Lu, Tun" sort="Lu, Tun" uniqKey="Lu T" first="Tun" last="Lu">Tun Lu</name><affiliation wicri:level="1"><nlm:affiliation>Bioengineering and Bioscience Department, Fuzhou University, Fuzhou 350108, China. Electronic address: lvtun@yahoo.com.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Bioengineering and Bioscience Department, Fuzhou University, Fuzhou 350108</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author><author><name sortKey="Wang, Shihua" sort="Wang, Shihua" uniqKey="Wang S" first="Shihua" last="Wang">Shihua Wang</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China. Electronic address: wshyyl@sina.com.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002</wicri:regionArea><placeName><settlement type="city">Fuzhou</settlement><region type="province">Fujian</region></placeName></affiliation></author></analytic><series><title level="j">Toxicon : official journal of the International Society on Toxinology</title><idno type="eISSN">1879-3150</idno><imprint><date when="2014" type="published">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence (MeSH)</term><term>Antibodies, Neutralizing (chemistry)</term><term>Antibody Affinity (MeSH)</term><term>Base Sequence (MeSH)</term><term>Binding Sites (MeSH)</term><term>Models, Molecular (MeSH)</term><term>Molecular Sequence Data (MeSH)</term><term>Peptide Library (MeSH)</term><term>Point Mutation (MeSH)</term><term>Sequence Analysis, Protein (MeSH)</term><term>Single-Chain Antibodies (chemistry)</term><term>Tetrodotoxin (immunology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Affinité des anticorps (MeSH)</term><term>Analyse de séquence de protéine (MeSH)</term><term>Anticorps neutralisants (composition chimique)</term><term>Anticorps à chaîne unique (composition chimique)</term><term>Banque de peptides (MeSH)</term><term>Données de séquences moléculaires (MeSH)</term><term>Modèles moléculaires (MeSH)</term><term>Mutation ponctuelle (MeSH)</term><term>Sites de fixation (MeSH)</term><term>Séquence d'acides aminés (MeSH)</term><term>Séquence nucléotidique (MeSH)</term><term>Tétrodotoxine (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Antibodies, Neutralizing</term><term>Single-Chain Antibodies</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Tetrodotoxin</term></keywords><keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Anticorps neutralisants</term><term>Anticorps à chaîne unique</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Tétrodotoxine</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Antibody Affinity</term><term>Base Sequence</term><term>Binding Sites</term><term>Models, Molecular</term><term>Molecular Sequence Data</term><term>Peptide Library</term><term>Point Mutation</term><term>Sequence Analysis, Protein</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Affinité des anticorps</term><term>Analyse de séquence de protéine</term><term>Banque de peptides</term><term>Données de séquences moléculaires</term><term>Modèles moléculaires</term><term>Mutation ponctuelle</term><term>Sites de fixation</term><term>Séquence d'acides aminés</term><term>Séquence nucléotidique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Tetrodotoxin (TTX) is a small molecular weight neurotoxin that occludes voltage-gated sodium channels in nerve and muscle tissue, resulting in respiratory paralysis and death. A high affinity antibody that can neutralize the toxicity of TTX is still lacking, so it is very important to prepare an antibody for TTX therapy and detection. In the present study, a chemical method was used to prepare the tetrodotoxin complete antigen, and a small amount, repeatedly immunity way was carried to immunize 4 mice. The amplified genes encoding monoclonal antibodies against TTX were used to construct the phage display library. After six rounds of biopanning, an antibody named scFv-T53 was characterized from clones showing high affinity and specific to TTX, and its affinity constant was 1.1 × 10(6) L/mol. Three dimensional structure of the scFv-T53 was constructed by computer modeling, and TTX was docked to the scFv-T53 model to obtain the structure of the binding complex. Two predicted essential amino acids, K183 and I189, were mutated to verify the theoretical model. Both mutants lost binding activity significantly against TTX as predicted by the theoretical model. Hence, the above results will be useful for screening the high affinity anti-TTX scFv mutants.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">24613546</PMID><DateCompleted><Year>2015</Year><Month>01</Month><Day>09</Day></DateCompleted><DateRevised><Year>2014</Year><Month>04</Month><Day>11</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1879-3150</ISSN><JournalIssue CitedMedium="Internet"><Volume>83</Volume><PubDate><Year>2014</Year><Month>Jun</Month></PubDate></JournalIssue><Title>Toxicon : official journal of the International Society on Toxinology</Title><ISOAbbreviation>Toxicon</ISOAbbreviation></Journal><ArticleTitle>Construction of a single chain variable fragment antibody (scFv) against tetrodotoxin (TTX) and its interaction with TTX.</ArticleTitle><Pagination><MedlinePgn>22-34</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.toxicon.2014.02.021</ELocationID><ELocationID EIdType="pii" ValidYN="Y">S0041-0101(14)00073-7</ELocationID><Abstract><AbstractText>Tetrodotoxin (TTX) is a small molecular weight neurotoxin that occludes voltage-gated sodium channels in nerve and muscle tissue, resulting in respiratory paralysis and death. A high affinity antibody that can neutralize the toxicity of TTX is still lacking, so it is very important to prepare an antibody for TTX therapy and detection. In the present study, a chemical method was used to prepare the tetrodotoxin complete antigen, and a small amount, repeatedly immunity way was carried to immunize 4 mice. The amplified genes encoding monoclonal antibodies against TTX were used to construct the phage display library. After six rounds of biopanning, an antibody named scFv-T53 was characterized from clones showing high affinity and specific to TTX, and its affinity constant was 1.1 × 10(6) L/mol. Three dimensional structure of the scFv-T53 was constructed by computer modeling, and TTX was docked to the scFv-T53 model to obtain the structure of the binding complex. Two predicted essential amino acids, K183 and I189, were mutated to verify the theoretical model. Both mutants lost binding activity significantly against TTX as predicted by the theoretical model. Hence, the above results will be useful for screening the high affinity anti-TTX scFv mutants.</AbstractText><CopyrightInformation>Copyright © 2014 Elsevier Ltd. All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Rongzhi</ForeName><Initials>R</Initials><AffiliationInfo><Affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Huang</LastName><ForeName>Ailing</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>Bioengineering and Bioscience Department, Fuzhou University, Fuzhou 350108, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Liu</LastName><ForeName>Licai</ForeName><Initials>L</Initials><AffiliationInfo><Affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Xiang</LastName><ForeName>Shuangshuang</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Xiufeng</ForeName><Initials>X</Initials><AffiliationInfo><Affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ling</LastName><ForeName>Sumei</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Lei</ForeName><Initials>L</Initials><AffiliationInfo><Affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Lu</LastName><ForeName>Tun</ForeName><Initials>T</Initials><AffiliationInfo><Affiliation>Bioengineering and Bioscience Department, Fuzhou University, Fuzhou 350108, China. Electronic address: lvtun@yahoo.com.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Shihua</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Key Laboratory of Pathogenic Fungi and Mycotxins of Fujian Province, The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China. Electronic address: wshyyl@sina.com.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2014</Year><Month>03</Month><Day>05</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Toxicon</MedlineTA><NlmUniqueID>1307333</NlmUniqueID><ISSNLinking>0041-0101</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D057134">Antibodies, Neutralizing</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D019151">Peptide Library</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D057127">Single-Chain Antibodies</NameOfSubstance></Chemical><Chemical><RegistryNumber>4368-28-9</RegistryNumber><NameOfSubstance UI="D013779">Tetrodotoxin</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000595" MajorTopicYN="N">Amino Acid Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D057134" MajorTopicYN="N">Antibodies, Neutralizing</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D000915" MajorTopicYN="N">Antibody Affinity</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001483" MajorTopicYN="N">Base Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001665" MajorTopicYN="N">Binding Sites</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008958" MajorTopicYN="N">Models, Molecular</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008969" MajorTopicYN="N">Molecular Sequence Data</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D019151" MajorTopicYN="N">Peptide Library</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D017354" MajorTopicYN="N">Point Mutation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D020539" MajorTopicYN="N">Sequence Analysis, Protein</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D057127" MajorTopicYN="N">Single-Chain Antibodies</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013779" MajorTopicYN="N">Tetrodotoxin</DescriptorName><QualifierName UI="Q000276" MajorTopicYN="Y">immunology</QualifierName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Computer modeling</Keyword><Keyword MajorTopicYN="N">Mutation</Keyword><Keyword MajorTopicYN="N">Tetrodotoxin (TTX)</Keyword><Keyword MajorTopicYN="N">scFv</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2013</Year><Month>11</Month><Day>24</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2014</Year><Month>02</Month><Day>20</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2014</Year><Month>02</Month><Day>25</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2014</Year><Month>3</Month><Day>12</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2014</Year><Month>3</Month><Day>13</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2015</Year><Month>1</Month><Day>13</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">24613546</ArticleId><ArticleId IdType="pii">S0041-0101(14)00073-7</ArticleId><ArticleId IdType="doi">10.1016/j.toxicon.2014.02.021</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>République populaire de Chine</li></country><region><li>Fujian</li></region><settlement><li>Fuzhou</li></settlement></list><tree><country name="République populaire de Chine"><region name="Fujian"><name sortKey="Wang, Rongzhi" sort="Wang, Rongzhi" uniqKey="Wang R" first="Rongzhi" last="Wang">Rongzhi Wang</name></region><name sortKey="Huang, Ailing" sort="Huang, Ailing" uniqKey="Huang A" first="Ailing" last="Huang">Ailing Huang</name><name sortKey="Li, Xiufeng" sort="Li, Xiufeng" uniqKey="Li X" first="Xiufeng" last="Li">Xiufeng Li</name><name sortKey="Ling, Sumei" sort="Ling, Sumei" uniqKey="Ling S" first="Sumei" last="Ling">Sumei Ling</name><name sortKey="Liu, Licai" sort="Liu, Licai" uniqKey="Liu L" first="Licai" last="Liu">Licai Liu</name><name sortKey="Lu, Tun" sort="Lu, Tun" uniqKey="Lu T" first="Tun" last="Lu">Tun Lu</name><name sortKey="Wang, Lei" sort="Wang, Lei" uniqKey="Wang L" first="Lei" last="Wang">Lei Wang</name><name sortKey="Wang, Shihua" sort="Wang, Shihua" uniqKey="Wang S" first="Shihua" last="Wang">Shihua Wang</name><name sortKey="Xiang, Shuangshuang" sort="Xiang, Shuangshuang" uniqKey="Xiang S" first="Shuangshuang" last="Xiang">Shuangshuang Xiang</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Bois/explor/BiopestPeptidV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 001B50 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 001B50 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Bois
|area= BiopestPeptidV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= pubmed:24613546
|texte= Construction of a single chain variable fragment antibody (scFv) against tetrodotoxin (TTX) and its interaction with TTX.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:24613546" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \
| NlmPubMed2Wicri -a BiopestPeptidV1
| This area was generated with Dilib version V0.6.38. |  |