Título:	Design and development of multiepitope chimeric antigens by bioinformatic and bacterial based recombinant expression methods, with potential application for bovine tuberculosis serodiagnosis.
Autor:	BATISTA, C. M. VESCHI, J. L. A. SOUZA, V. F. de FOTI, L. CHAPAVAL, L.
Afiliación:	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.
Año:	2024
Referencia:	Veterinary Immunology and Immunopathology, v. 269, 110729, 2024.
Descripción:	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.
Thesagro:	Tuberculose Elisa Bovino Doença Animal
NAL Thesaurus:	Mycobacterium bovis BCG Bovine tuberculosis
Palabras clave:	Tuberculose bovina Antígenos quiméricos multiepítopos Imunoensaios
DOI:	https://doi.org/10.1016/j.vetimm.2024.110729
Tipo de Material:	Artigo de periódico
Acceso:	openAccess
Aparece en las colecciones:	Artigo em periódico indexado (CPATSA)