Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis.

Detalhes bibliográficos
Autor(a) principal: BATISTA, C. M.
Data de Publicação: 2024
Outros Autores: VESCHI, J. L. A., SOUZA, V. F. de, FOTI, L., CHAPAVAL, L.
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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spelling 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
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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)
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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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