A novel B cell epitope in cold-shock DEAD-box protein A from Mycobacterium tuberculosis.
Identifieur interne : 001D30 ( PubMed/Corpus ); précédent : 001D29; suivant : 001D31A novel B cell epitope in cold-shock DEAD-box protein A from Mycobacterium tuberculosis.
Auteurs : Huanan Wang ; Ting Zhu ; Shenye Yu ; Huifang Liu ; Xiumei Wang ; Liping Chen ; Wei Si ; Hai Pang ; Siguo LiuSource :
- Research in veterinary science [ 1532-2661 ] ; 2013.
English descriptors
- KwdEn :
- Animals, Antibodies, Monoclonal (immunology), Antibody Specificity (immunology), Blotting, Western, Cell Line, Cloning, Molecular, DEAD-box RNA Helicases (genetics), DEAD-box RNA Helicases (immunology), Enzyme-Linked Immunosorbent Assay, Epitopes, B-Lymphocyte (immunology), Escherichia coli (genetics), Hybridomas, Mice, Mycobacterium tuberculosis (genetics), Mycobacterium tuberculosis (immunology), Recombinant Proteins (genetics), Recombinant Proteins (immunology), Staphylococcal Protein A (genetics), Staphylococcal Protein A (immunology).
- MESH :
- chemical , genetics : DEAD-box RNA Helicases, Recombinant Proteins, Staphylococcal Protein A.
- chemical , immunology : Antibodies, Monoclonal, DEAD-box RNA Helicases, Epitopes, B-Lymphocyte, Recombinant Proteins, Staphylococcal Protein A.
- genetics : Escherichia coli, Mycobacterium tuberculosis.
- immunology : Antibody Specificity, Mycobacterium tuberculosis.
- Animals, Blotting, Western, Cell Line, Cloning, Molecular, Enzyme-Linked Immunosorbent Assay, Hybridomas, Mice.
Abstract
In this study, a hybridoma-based technique and phage display technology were used to obtain mouse monoclonal antibodies (mAb) against cold-shock DEAD-box protein A (CsdA) from Mycobacterium tuberculosis and to determine the location of the relevant epitope. One highly specific mAb, named A3G5, was developed against the recombinant CsdA protein (rCsdA) and could detect rCsdA protein in enzyme-linked immunosorbent assays (ELISA) and Western blot assays. By screening a phage displayed library of random 12-mers (Ph.D.-12), 10 positive phage clones were randomly selected after three rounds of bio-panning and identified by ELISA. Eight of these clones were sequenced, and their amino acid sequences were deduced. One B-cell epitope (-APDPPLSRR-) in the rCsdA protein was identified with mAb A3G5. A synthetic peptide (-MAPDPPLSRR-) (Cpep) matched well with the CsdA sequence at 443-451 aa and was confirmed by affinity ELISA, competitive inhibition assays and the development of an immune response in mice. These results may be of great potential value in the further analysis of the function and structure of the CsdA protein from M. tuberculosis.
DOI: 10.1016/j.rvsc.2012.11.013
PubMed: 23261158
Links to Exploration step
pubmed:23261158Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">A novel B cell epitope in cold-shock DEAD-box protein A from Mycobacterium tuberculosis.</title><author><name sortKey="Wang, Huanan" sort="Wang, Huanan" uniqKey="Wang H" first="Huanan" last="Wang">Huanan Wang</name><affiliation><nlm:affiliation>Division of Bacterial Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 15000, PR China.</nlm:affiliation></affiliation></author><author><name sortKey="Zhu, Ting" sort="Zhu, Ting" uniqKey="Zhu T" first="Ting" last="Zhu">Ting Zhu</name></author><author><name sortKey="Yu, Shenye" sort="Yu, Shenye" uniqKey="Yu S" first="Shenye" last="Yu">Shenye Yu</name></author><author><name sortKey="Liu, Huifang" sort="Liu, Huifang" uniqKey="Liu H" first="Huifang" last="Liu">Huifang Liu</name></author><author><name sortKey="Wang, Xiumei" sort="Wang, Xiumei" uniqKey="Wang X" first="Xiumei" last="Wang">Xiumei Wang</name></author><author><name sortKey="Chen, Liping" sort="Chen, Liping" uniqKey="Chen L" first="Liping" last="Chen">Liping Chen</name></author><author><name sortKey="Si, Wei" sort="Si, Wei" uniqKey="Si W" first="Wei" last="Si">Wei Si</name></author><author><name sortKey="Pang, Hai" sort="Pang, Hai" uniqKey="Pang H" first="Hai" last="Pang">Hai Pang</name></author><author><name sortKey="Liu, Siguo" sort="Liu, Siguo" uniqKey="Liu S" first="Siguo" last="Liu">Siguo Liu</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2013">2013</date><idno type="RBID">pubmed:23261158</idno><idno type="pmid">23261158</idno><idno type="doi">10.1016/j.rvsc.2012.11.013</idno><idno type="wicri:Area/PubMed/Corpus">001D30</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001D30</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">A novel B cell epitope in cold-shock DEAD-box protein A from Mycobacterium tuberculosis.</title><author><name sortKey="Wang, Huanan" sort="Wang, Huanan" uniqKey="Wang H" first="Huanan" last="Wang">Huanan Wang</name><affiliation><nlm:affiliation>Division of Bacterial Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 15000, PR China.</nlm:affiliation></affiliation></author><author><name sortKey="Zhu, Ting" sort="Zhu, Ting" uniqKey="Zhu T" first="Ting" last="Zhu">Ting Zhu</name></author><author><name sortKey="Yu, Shenye" sort="Yu, Shenye" uniqKey="Yu S" first="Shenye" last="Yu">Shenye Yu</name></author><author><name sortKey="Liu, Huifang" sort="Liu, Huifang" uniqKey="Liu H" first="Huifang" last="Liu">Huifang Liu</name></author><author><name sortKey="Wang, Xiumei" sort="Wang, Xiumei" uniqKey="Wang X" first="Xiumei" last="Wang">Xiumei Wang</name></author><author><name sortKey="Chen, Liping" sort="Chen, Liping" uniqKey="Chen L" first="Liping" last="Chen">Liping Chen</name></author><author><name sortKey="Si, Wei" sort="Si, Wei" uniqKey="Si W" first="Wei" last="Si">Wei Si</name></author><author><name sortKey="Pang, Hai" sort="Pang, Hai" uniqKey="Pang H" first="Hai" last="Pang">Hai Pang</name></author><author><name sortKey="Liu, Siguo" sort="Liu, Siguo" uniqKey="Liu S" first="Siguo" last="Liu">Siguo Liu</name></author></analytic><series><title level="j">Research in veterinary science</title><idno type="eISSN">1532-2661</idno><imprint><date when="2013" type="published">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Monoclonal (immunology)</term><term>Antibody Specificity (immunology)</term><term>Blotting, Western</term><term>Cell Line</term><term>Cloning, Molecular</term><term>DEAD-box RNA Helicases (genetics)</term><term>DEAD-box RNA Helicases (immunology)</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Epitopes, B-Lymphocyte (immunology)</term><term>Escherichia coli (genetics)</term><term>Hybridomas</term><term>Mice</term><term>Mycobacterium tuberculosis (genetics)</term><term>Mycobacterium tuberculosis (immunology)</term><term>Recombinant Proteins (genetics)</term><term>Recombinant Proteins (immunology)</term><term>Staphylococcal Protein A (genetics)</term><term>Staphylococcal Protein A (immunology)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DEAD-box RNA Helicases</term><term>Recombinant Proteins</term><term>Staphylococcal Protein A</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Monoclonal</term><term>DEAD-box RNA Helicases</term><term>Epitopes, B-Lymphocyte</term><term>Recombinant Proteins</term><term>Staphylococcal Protein A</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Escherichia coli</term><term>Mycobacterium tuberculosis</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Antibody Specificity</term><term>Mycobacterium tuberculosis</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Blotting, Western</term><term>Cell Line</term><term>Cloning, Molecular</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Hybridomas</term><term>Mice</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">In this study, a hybridoma-based technique and phage display technology were used to obtain mouse monoclonal antibodies (mAb) against cold-shock DEAD-box protein A (CsdA) from Mycobacterium tuberculosis and to determine the location of the relevant epitope. One highly specific mAb, named A3G5, was developed against the recombinant CsdA protein (rCsdA) and could detect rCsdA protein in enzyme-linked immunosorbent assays (ELISA) and Western blot assays. By screening a phage displayed library of random 12-mers (Ph.D.-12), 10 positive phage clones were randomly selected after three rounds of bio-panning and identified by ELISA. Eight of these clones were sequenced, and their amino acid sequences were deduced. One B-cell epitope (-APDPPLSRR-) in the rCsdA protein was identified with mAb A3G5. A synthetic peptide (-MAPDPPLSRR-) (Cpep) matched well with the CsdA sequence at 443-451 aa and was confirmed by affinity ELISA, competitive inhibition assays and the development of an immune response in mice. These results may be of great potential value in the further analysis of the function and structure of the CsdA protein from M. tuberculosis.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">23261158</PMID><DateCompleted><Year>2013</Year><Month>12</Month><Day>09</Day></DateCompleted><DateRevised><Year>2013</Year><Month>04</Month><Day>09</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1532-2661</ISSN><JournalIssue CitedMedium="Internet"><Volume>94</Volume><Issue>3</Issue><PubDate><Year>2013</Year><Month>Jun</Month></PubDate></JournalIssue><Title>Research in veterinary science</Title><ISOAbbreviation>Res. Vet. Sci.</ISOAbbreviation></Journal><ArticleTitle>A novel B cell epitope in cold-shock DEAD-box protein A from Mycobacterium tuberculosis.</ArticleTitle><Pagination><MedlinePgn>406-12</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.rvsc.2012.11.013</ELocationID><ELocationID EIdType="pii" ValidYN="Y">S0034-5288(12)00346-3</ELocationID><Abstract><AbstractText>In this study, a hybridoma-based technique and phage display technology were used to obtain mouse monoclonal antibodies (mAb) against cold-shock DEAD-box protein A (CsdA) from Mycobacterium tuberculosis and to determine the location of the relevant epitope. One highly specific mAb, named A3G5, was developed against the recombinant CsdA protein (rCsdA) and could detect rCsdA protein in enzyme-linked immunosorbent assays (ELISA) and Western blot assays. By screening a phage displayed library of random 12-mers (Ph.D.-12), 10 positive phage clones were randomly selected after three rounds of bio-panning and identified by ELISA. Eight of these clones were sequenced, and their amino acid sequences were deduced. One B-cell epitope (-APDPPLSRR-) in the rCsdA protein was identified with mAb A3G5. A synthetic peptide (-MAPDPPLSRR-) (Cpep) matched well with the CsdA sequence at 443-451 aa and was confirmed by affinity ELISA, competitive inhibition assays and the development of an immune response in mice. These results may be of great potential value in the further analysis of the function and structure of the CsdA protein from M. tuberculosis.</AbstractText><CopyrightInformation>Crown Copyright © 2012. Published by Elsevier India Pvt Ltd. All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Huanan</ForeName><Initials>H</Initials><AffiliationInfo><Affiliation>Division of Bacterial Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 15000, PR China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhu</LastName><ForeName>Ting</ForeName><Initials>T</Initials></Author><Author ValidYN="Y"><LastName>Yu</LastName><ForeName>Shenye</ForeName><Initials>S</Initials></Author><Author ValidYN="Y"><LastName>Liu</LastName><ForeName>Huifang</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Xiumei</ForeName><Initials>X</Initials></Author><Author ValidYN="Y"><LastName>Chen</LastName><ForeName>Liping</ForeName><Initials>L</Initials></Author><Author ValidYN="Y"><LastName>Si</LastName><ForeName>Wei</ForeName><Initials>W</Initials></Author><Author ValidYN="Y"><LastName>Pang</LastName><ForeName>Hai</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Liu</LastName><ForeName>Siguo</ForeName><Initials>S</Initials></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>2012</Year><Month>12</Month><Day>20</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Res Vet Sci</MedlineTA><NlmUniqueID>0401300</NlmUniqueID><ISSNLinking>0034-5288</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000911">Antibodies, Monoclonal</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D018985">Epitopes, B-Lymphocyte</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D013205">Staphylococcal Protein A</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.6.4.13</RegistryNumber><NameOfSubstance UI="D053487">DEAD-box RNA Helicases</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000911" MajorTopicYN="N">Antibodies, Monoclonal</DescriptorName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D000918" MajorTopicYN="N">Antibody Specificity</DescriptorName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D015153" MajorTopicYN="N">Blotting, Western</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002460" MajorTopicYN="N">Cell Line</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D003001" MajorTopicYN="N">Cloning, Molecular</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D053487" MajorTopicYN="N">DEAD-box RNA Helicases</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004797" MajorTopicYN="N">Enzyme-Linked Immunosorbent Assay</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018985" MajorTopicYN="N">Epitopes, B-Lymphocyte</DescriptorName><QualifierName UI="Q000276" MajorTopicYN="Y">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004926" MajorTopicYN="N">Escherichia coli</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006825" MajorTopicYN="N">Hybridomas</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D051379" MajorTopicYN="N">Mice</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D009169" MajorTopicYN="N">Mycobacterium tuberculosis</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013205" MajorTopicYN="N">Staphylococcal Protein A</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2012</Year><Month>05</Month><Day>26</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2012</Year><Month>11</Month><Day>19</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2012</Year><Month>11</Month><Day>22</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2012</Year><Month>12</Month><Day>25</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2012</Year><Month>12</Month><Day>25</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2013</Year><Month>12</Month><Day>16</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">23261158</ArticleId><ArticleId IdType="pii">S0034-5288(12)00346-3</ArticleId><ArticleId 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