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| 4. |  | SANTOS, D. C. dos; WENDLING, I. Estudos para definição de meios de cultura e métodos de desinfestação de explantes de plantas adultas de erva-mate (Ilex paraguariensis St. Hill). In: CONGRESSO SUL-AMERICANO DA ERVA-MATE, 3., 2003, Chapecó. Anais. [Chapecó]: EPAGRI, 2003. s.3-12. 1 CD-ROM. Secão: Conservação, Melhoramento e Multiplicação. Feira do Agronegócio da Erva-mate, 1., 2003, Chapecó. Integrar para promover o agronegócio da erva-mate.| Biblioteca(s): Embrapa Florestas. |
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| 7. | | SOUZA JÚNIOR, L.; WENDLING, I; SANTOS, D. C. dos. Fitorreguladores de crescimento no alongamento in vitro de Pinus greggii. In: EVENTO DE INICIAÇÃO CIENTÍFICA DA EMBRAPA FLORESTAS, 2., 2003, Colombo. Anais. Colombo: Embrapa Florestas, 2003. 027A. 1 Cd-Rom. (Embrapa Florestas. Documentos, 86). Organizado por Patricia Póvoa de Mattos, Celso Garcia Auer, Rejane Stumpf Sberze, Katia Regina Pichelli e Paulo César Botosso.| Biblioteca(s): Embrapa Florestas. |
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| 15. | | SANTOS, D. C. dos; WENDLING, I.; GROSSI, F. Influência de diferentes relações de reguladores de crescimento no alongamento in vitro de microestacas de Grevillea robusta (Cunn.). In: EVENTO DE INICIAÇÃO CIENTÍFICA DA EMBRAPA FLORESTAS, 1., 2002, Colombo. Anais. Colombo: Embrapa Florestas, 2002. 1 CD-ROM. (Embrapa Florestas. Documentos, 70). Organizado por Patrícia Póvoa de Mattos, Celso Garcia Auer, Paulo César Botosso e Rejane Stumpf Sberze.| Biblioteca(s): Embrapa Florestas. |
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Registro Completo
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Biblioteca(s): |
Embrapa Semiárido. |
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Data corrente: |
09/07/2024 |
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Data da última atualização: |
09/07/2024 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Circulação/Nível: |
A - 1 |
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Autoria: |
BATISTA, C. M.; VESCHI, J. L. A.; SOUZA, V. F. de; FOTI, L.; CHAPAVAL, L. |
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Afiliação: |
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. |
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Título: |
Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis. |
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Ano de publicação: |
2024 |
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Fonte/Imprenta: |
Veterinary Immunology and Immunopathology, v. 269, 110729, 2024. |
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DOI: |
https://doi.org/10.1016/j.vetimm.2024.110729 |
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Idioma: |
Inglês |
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Conteúdo: |
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. MenosBovine 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... Mostrar Tudo |
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Palavras-Chave: |
Antígenos quiméricos multiepítopos; Imunoensaios; Tuberculose bovina. |
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Thesagro: |
Bovino; Doença Animal; Elisa; Tuberculose. |
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Thesaurus NAL: |
Bovine tuberculosis; Mycobacterium bovis BCG. |
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Categoria do assunto: |
L Ciência Animal e Produtos de Origem Animal |
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URL: |
https://www.alice.cnptia.embrapa.br/alice/bitstream/doc/1165582/1/Design-and-development-of-multiepitope-chimeric-antigens.pdf
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Marc: |
LEADER 02609naa a2200289 a 4500
001 2165582
005 2024-07-09
008 2024 bl uuuu u00u1 u #d
024 7 $ahttps://doi.org/10.1016/j.vetimm.2024.110729$2DOI
100 1 $aBATISTA, C. M.
245 $aDesign and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis.$h[electronic resource]
260 $c2024
520 $aBovine 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.
650 $aBovine tuberculosis
650 $aMycobacterium bovis BCG
650 $aBovino
650 $aDoença Animal
650 $aElisa
650 $aTuberculose
653 $aAntígenos quiméricos multiepítopos
653 $aImunoensaios
653 $aTuberculose bovina
700 1 $aVESCHI, J. L. A.
700 1 $aSOUZA, V. F. de
700 1 $aFOTI, L.
700 1 $aCHAPAVAL, L.
773 $tVeterinary Immunology and Immunopathology$gv. 269, 110729, 2024.
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