A novel B cell epitope in cold-shock DEAD-box protein A from Mycobacterium tuberculosis.
Identifieur interne : 002066 ( Main/Exploration ); précédent : 002065; suivant : 002067A novel B cell epitope in cold-shock DEAD-box protein A from Mycobacterium tuberculosis.
Auteurs : Huanan Wang [République populaire de Chine] ; Ting Zhu ; Shenye Yu ; Huifang Liu ; Xiumei Wang ; Liping Chen ; Wei Si ; Hai Pang ; Siguo LiuSource :
- Research in veterinary science [ 1532-2661 ] ; 2013.
Descripteurs français
- KwdFr :
- Animaux, Anticorps monoclonaux (immunologie), Clonage moléculaire, DEAD-box RNA helicases (génétique), DEAD-box RNA helicases (immunologie), Déterminants antigéniques des lymphocytes B (immunologie), Escherichia coli (génétique), Hybridomes, Lignée cellulaire, Mycobacterium tuberculosis (génétique), Mycobacterium tuberculosis (immunologie), Protéine A staphylococcique (génétique), Protéine A staphylococcique (immunologie), Protéines recombinantes (génétique), Protéines recombinantes (immunologie), Souris, Spécificité des anticorps (immunologie), Technique de Western, Test ELISA.
- MESH :
- génétique : DEAD-box RNA helicases, Escherichia coli, Mycobacterium tuberculosis, Protéine A staphylococcique, Protéines recombinantes.
- immunologie : Anticorps monoclonaux, DEAD-box RNA helicases, Déterminants antigéniques des lymphocytes B, Mycobacterium tuberculosis, Protéine A staphylococcique, Protéines recombinantes, Spécificité des anticorps.
- Animaux, Clonage moléculaire, Hybridomes, Lignée cellulaire, Souris, Technique de Western, Test ELISA.
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
Affiliations:
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tuberculosis.</title><author><name sortKey="Wang, Huanan" sort="Wang, Huanan" uniqKey="Wang H" first="Huanan" last="Wang">Huanan Wang</name><affiliation wicri:level="1"><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><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Division of Bacterial Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 15000</wicri:regionArea><wicri:noRegion>Harbin 15000</wicri:noRegion></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="KwdFr" xml:lang="fr"><term>Animaux</term><term>Anticorps monoclonaux (immunologie)</term><term>Clonage moléculaire</term><term>DEAD-box RNA helicases (génétique)</term><term>DEAD-box RNA helicases (immunologie)</term><term>Déterminants antigéniques des lymphocytes B (immunologie)</term><term>Escherichia coli (génétique)</term><term>Hybridomes</term><term>Lignée cellulaire</term><term>Mycobacterium tuberculosis (génétique)</term><term>Mycobacterium tuberculosis (immunologie)</term><term>Protéine A staphylococcique (génétique)</term><term>Protéine A staphylococcique (immunologie)</term><term>Protéines recombinantes (génétique)</term><term>Protéines recombinantes (immunologie)</term><term>Souris</term><term>Spécificité des anticorps (immunologie)</term><term>Technique de Western</term><term>Test ELISA</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="génétique" xml:lang="fr"><term>DEAD-box RNA helicases</term><term>Escherichia coli</term><term>Mycobacterium tuberculosis</term><term>Protéine A staphylococcique</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps monoclonaux</term><term>DEAD-box RNA helicases</term><term>Déterminants antigéniques des lymphocytes B</term><term>Mycobacterium tuberculosis</term><term>Protéine A staphylococcique</term><term>Protéines recombinantes</term><term>Spécificité des anticorps</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><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Clonage moléculaire</term><term>Hybridomes</term><term>Lignée cellulaire</term><term>Souris</term><term>Technique de Western</term><term>Test ELISA</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><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><noCountry><name sortKey="Chen, Liping" sort="Chen, Liping" uniqKey="Chen L" first="Liping" last="Chen">Liping Chen</name><name sortKey="Liu, Huifang" sort="Liu, Huifang" uniqKey="Liu H" first="Huifang" last="Liu">Huifang Liu</name><name sortKey="Liu, Siguo" sort="Liu, Siguo" uniqKey="Liu S" first="Siguo" last="Liu">Siguo Liu</name><name sortKey="Pang, Hai" sort="Pang, Hai" uniqKey="Pang H" first="Hai" last="Pang">Hai Pang</name><name sortKey="Si, Wei" sort="Si, Wei" uniqKey="Si W" first="Wei" last="Si">Wei Si</name><name sortKey="Wang, Xiumei" sort="Wang, Xiumei" uniqKey="Wang X" first="Xiumei" last="Wang">Xiumei Wang</name><name sortKey="Yu, Shenye" sort="Yu, Shenye" uniqKey="Yu S" first="Shenye" last="Yu">Shenye Yu</name><name sortKey="Zhu, Ting" sort="Zhu, Ting" uniqKey="Zhu T" first="Ting" last="Zhu">Ting Zhu</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Wang, Huanan" sort="Wang, Huanan" uniqKey="Wang H" first="Huanan" last="Wang">Huanan Wang</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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