Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis.
Autor(a) principal: | |
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Data de Publicação: | 2024 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
Texto Completo: | http://www.alice.cnptia.embrapa.br/alice/handle/doc/1165582 https://doi.org/10.1016/j.vetimm.2024.110729 |
Resumo: | Bovine tuberculosis (bTB), which is caused by Mycobacterium bovis, is a single health concern, which causes economic losses, is a sanitary barrier and is a zoonotic concern. The golden-pattern intradermic tests have low sensitivity of about 50%. To fix this sensitivity problem, immunoassays could be a powerful tool. However, few studies produced antigens for bTB immunoassays, which needs improvements. Aim of this study was to produce multiepitope chimeric antigens (MCA) to use for bTB diagnosis. To achieve MCA design and development, extensive bibliographic search, antigenic epitope prediction, specificity, hydrophobicity, and 3D structure modeling analyses were performed, as well as cloning, expression and purification. Seven epitopes from four different target proteins (MPB-70, MPB-83, ESAT-6 and GroEL) were combined in five chimeras containing five repetitions of each epitope to enhance antibodies affinity. 3D predicted models revealed that all chimeras have a high percentage of disorder, which could enhance antibody recognition, although taking to protein instability. Each chimera was cloned into pET28a (+) expression plasmids and expressed in six Escherichia coli expression strains. Chimeras 3, 4 and 5 could be solubilized in 8 M urea and purified by ion exchange affinity chromatography. Against bTB positive and negative sera, purified chimera 5 had the best results in indirect dot blot and ELISA, as well as in lateral flow dot blot immunoassay. In conclusion, chimera 5, an MPB-83 containing MCA, could be used for further studies, aimed to develop a serologic or rapid test for bTB diagnosis. |
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Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis.Tuberculose bovinaAntígenos quiméricos multiepítoposImunoensaiosTuberculoseElisaBovinoDoença AnimalMycobacterium bovis BCGBovine tuberculosisBovine tuberculosis (bTB), which is caused by Mycobacterium bovis, is a single health concern, which causes economic losses, is a sanitary barrier and is a zoonotic concern. The golden-pattern intradermic tests have low sensitivity of about 50%. To fix this sensitivity problem, immunoassays could be a powerful tool. However, few studies produced antigens for bTB immunoassays, which needs improvements. Aim of this study was to produce multiepitope chimeric antigens (MCA) to use for bTB diagnosis. To achieve MCA design and development, extensive bibliographic search, antigenic epitope prediction, specificity, hydrophobicity, and 3D structure modeling analyses were performed, as well as cloning, expression and purification. Seven epitopes from four different target proteins (MPB-70, MPB-83, ESAT-6 and GroEL) were combined in five chimeras containing five repetitions of each epitope to enhance antibodies affinity. 3D predicted models revealed that all chimeras have a high percentage of disorder, which could enhance antibody recognition, although taking to protein instability. Each chimera was cloned into pET28a (+) expression plasmids and expressed in six Escherichia coli expression strains. Chimeras 3, 4 and 5 could be solubilized in 8 M urea and purified by ion exchange affinity chromatography. Against bTB positive and negative sera, purified chimera 5 had the best results in indirect dot blot and ELISA, as well as in lateral flow dot blot immunoassay. In conclusion, chimera 5, an MPB-83 containing MCA, could be used for further studies, aimed to develop a serologic or rapid test for bTB diagnosis.CASSIANO MARTIN BATISTA, INSTITUTO CARLOS CHAGAS; JOSIR LAINE APARECIDA VESCHI, CPATSA; VANESSA FELIPE DE SOUZA, CNPGC; LEONARDO FOTI, INSTITUTO CARLOS CHAGAS; LEA CHAPAVAL ANDRI, CPPSE.BATISTA, C. M.VESCHI, J. L. A.SOUZA, V. F. deFOTI, L.CHAPAVAL, L.2024-07-09T11:54:51Z2024-07-09T11:54:51Z2024-07-092024info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleVeterinary Immunology and Immunopathology, v. 269, 110729, 2024.http://www.alice.cnptia.embrapa.br/alice/handle/doc/1165582https://doi.org/10.1016/j.vetimm.2024.110729enginfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)instacron:EMBRAPA2024-07-09T11:54:51Zoai:www.alice.cnptia.embrapa.br:doc/1165582Repositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestcg-riaa@embrapa.bropendoar:21542024-07-09T11:54:51Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)false |
dc.title.none.fl_str_mv |
Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis. |
title |
Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis. |
spellingShingle |
Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis. BATISTA, C. M. Tuberculose bovina Antígenos quiméricos multiepítopos Imunoensaios Tuberculose Elisa Bovino Doença Animal Mycobacterium bovis BCG Bovine tuberculosis |
title_short |
Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis. |
title_full |
Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis. |
title_fullStr |
Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis. |
title_full_unstemmed |
Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis. |
title_sort |
Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis. |
author |
BATISTA, C. M. |
author_facet |
BATISTA, C. M. VESCHI, J. L. A. SOUZA, V. F. de FOTI, L. CHAPAVAL, L. |
author_role |
author |
author2 |
VESCHI, J. L. A. SOUZA, V. F. de FOTI, L. CHAPAVAL, L. |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
CASSIANO MARTIN BATISTA, INSTITUTO CARLOS CHAGAS; JOSIR LAINE APARECIDA VESCHI, CPATSA; VANESSA FELIPE DE SOUZA, CNPGC; LEONARDO FOTI, INSTITUTO CARLOS CHAGAS; LEA CHAPAVAL ANDRI, CPPSE. |
dc.contributor.author.fl_str_mv |
BATISTA, C. M. VESCHI, J. L. A. SOUZA, V. F. de FOTI, L. CHAPAVAL, L. |
dc.subject.por.fl_str_mv |
Tuberculose bovina Antígenos quiméricos multiepítopos Imunoensaios Tuberculose Elisa Bovino Doença Animal Mycobacterium bovis BCG Bovine tuberculosis |
topic |
Tuberculose bovina Antígenos quiméricos multiepítopos Imunoensaios Tuberculose Elisa Bovino Doença Animal Mycobacterium bovis BCG Bovine tuberculosis |
description |
Bovine tuberculosis (bTB), which is caused by Mycobacterium bovis, is a single health concern, which causes economic losses, is a sanitary barrier and is a zoonotic concern. The golden-pattern intradermic tests have low sensitivity of about 50%. To fix this sensitivity problem, immunoassays could be a powerful tool. However, few studies produced antigens for bTB immunoassays, which needs improvements. Aim of this study was to produce multiepitope chimeric antigens (MCA) to use for bTB diagnosis. To achieve MCA design and development, extensive bibliographic search, antigenic epitope prediction, specificity, hydrophobicity, and 3D structure modeling analyses were performed, as well as cloning, expression and purification. Seven epitopes from four different target proteins (MPB-70, MPB-83, ESAT-6 and GroEL) were combined in five chimeras containing five repetitions of each epitope to enhance antibodies affinity. 3D predicted models revealed that all chimeras have a high percentage of disorder, which could enhance antibody recognition, although taking to protein instability. Each chimera was cloned into pET28a (+) expression plasmids and expressed in six Escherichia coli expression strains. Chimeras 3, 4 and 5 could be solubilized in 8 M urea and purified by ion exchange affinity chromatography. Against bTB positive and negative sera, purified chimera 5 had the best results in indirect dot blot and ELISA, as well as in lateral flow dot blot immunoassay. In conclusion, chimera 5, an MPB-83 containing MCA, could be used for further studies, aimed to develop a serologic or rapid test for bTB diagnosis. |
publishDate |
2024 |
dc.date.none.fl_str_mv |
2024-07-09T11:54:51Z 2024-07-09T11:54:51Z 2024-07-09 2024 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
Veterinary Immunology and Immunopathology, v. 269, 110729, 2024. http://www.alice.cnptia.embrapa.br/alice/handle/doc/1165582 https://doi.org/10.1016/j.vetimm.2024.110729 |
identifier_str_mv |
Veterinary Immunology and Immunopathology, v. 269, 110729, 2024. |
url |
http://www.alice.cnptia.embrapa.br/alice/handle/doc/1165582 https://doi.org/10.1016/j.vetimm.2024.110729 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa) instacron:EMBRAPA |
instname_str |
Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
instacron_str |
EMBRAPA |
institution |
EMBRAPA |
reponame_str |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
collection |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
repository.name.fl_str_mv |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
repository.mail.fl_str_mv |
cg-riaa@embrapa.br |
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1817695714533179392 |