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Autor | Batista, Cassiano Martin | |
Autor | Veschi, Josir Laine | |
Autor | Souza, Vanessa Felipe de | |
Autor | Foti, Leonardo | |
Autor | Andri, Lea Chapaval | |
Fecha de acceso | 2024-03-08T19:49:40Z | |
Fecha de disponibilización | 2024-03-08T19:49:40Z | |
Fecha de publicación | 2024 | |
Referencia | BATISTA, Cassiano Martin et al. Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis. Veterinary Immunology and Immunopathology, v. 269, n. 110729, p. 1-7, 2024. | en_US |
ISSN | 0165-2427 | en_US |
URI | https://www.arca.fiocruz.br/handle/icict/62993 | |
Idioma | por | en_US |
Editor | Elsevier | en_US |
Derechos de autor | restricted access | en_US |
Palabras clave en Portugués | Antígenos Quiméricos Multiepítopos | en_US |
Palabras clave en Portugués | ELISA | en_US |
Título | Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis | en_US |
Tipo del documento | Article | en_US |
DOI | 10.1016/j.vetimm.2024.110729 | |
Resumen en Inglés | 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. | en_US |
Afiliación | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil / Empresa Brasileira de Pesquisa Agropecuária (Embrapa Pecuária Sudeste). São Carlos, SP, Brasil. | en_US |
Afiliación | Empresa Brasileira de Pesquisa Agropecuária (Embrapa Semiárido). Petrolina, PE, Brasil. | en_US |
Afiliación | Empresa Brasileira de Pesquisa Agropecuária (Embrapa Gado de Corte). Mato Grosso do Sul, MS, Brasil. | en_US |
Afiliación | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil. | en_US |
Afiliación | Empresa Brasileira de Pesquisa Agropecuária (Embrapa Pecuária Sudeste). São Carlos, SP, Brasil. | en_US |
Palavras clave en Inglês | Tuberculosis Bovine | en_US |
Palavras clave en Inglês | Diagnosis | en_US |
Palavras clave en Inglês | Multiepitope Chimeric Antigens | en_US |
Palavras clave en Inglês | Immunoassay | en_US |
Palavras clave en Inglês | Enzyme-Linked Immunosorbent Assay | en_US |
Palavras clave | Tuberculosis Bovina | en_US |
Palavras clave | Diagnóstico | en_US |
Palavras clave | Inmunoensayo | en_US |
Palavras clave | Ensayo de Inmunoadsorción Enzimática | en_US |
Palavras clave en Francês | Tuberculose bovine | en_US |
Palavras clave en Francês | Diagnostic | en_US |
Palavras clave en Francês | Dosage immunologique | en_US |
Palavras clave en Francês | Test ELISA | en_US |
DeCS | Tuberculose Bovina | en_US |
DeCS | Diagnóstico | en_US |
DeCS | Imunoensaio | en_US |
DeCS | Ensaio de Imunoadsorção Enzimática | en_US |