Production of recombinant NS1 protein and its possible use in encephalitic flavivirus differential diagnosis.

Autor: Lorch MS; Área de Virosis Emergentes y Zoonóticas (AVEZ), Laboratorio de Ingeniería Genética y Biología Celular y Molecular (LIGBCM), Departamento de Ciencia y Tecnología (DCyT), Universidad Nacional de Quilmes (UNQ), Roque Sáenz Peña 352, B1876BXD, Bernal, Buenos Aires, Argentina., Collado MS; Área de Virosis Emergentes y Zoonóticas (AVEZ), Laboratorio de Ingeniería Genética y Biología Celular y Molecular (LIGBCM), Departamento de Ciencia y Tecnología (DCyT), Universidad Nacional de Quilmes (UNQ), Roque Sáenz Peña 352, B1876BXD, Bernal, Buenos Aires, Argentina., Argüelles MH; Laboratorio de Inmunología y Virología (LIV), Departamento de Ciencia y Tecnología (DCyT), Universidad Nacional de Quilmes (UNQ), Roque Sáenz Peña 352, B1876BXD, Bernal, Buenos Aires, Argentina., Rota RP; Área de Virosis Emergentes y Zoonóticas (AVEZ), Laboratorio de Ingeniería Genética y Biología Celular y Molecular (LIGBCM), Departamento de Ciencia y Tecnología (DCyT), Universidad Nacional de Quilmes (UNQ), Roque Sáenz Peña 352, B1876BXD, Bernal, Buenos Aires, Argentina., Spinsanti LI; Laboratorio de Arenavirus y Arbovirus, Instituto de Virología 'Dr. J. M. Vanella', Facultad de Ciencias Médicas, Universidad Nacional de Córdoba (UNC), Enfermera Gordillo Gómez S/N., 5016, Ciudad Universitaria, Córdoba, Argentina., Lozano ME; Área de Virosis Emergentes y Zoonóticas (AVEZ), Laboratorio de Ingeniería Genética y Biología Celular y Molecular (LIGBCM), Departamento de Ciencia y Tecnología (DCyT), Universidad Nacional de Quilmes (UNQ), Roque Sáenz Peña 352, B1876BXD, Bernal, Buenos Aires, Argentina., Goñi SE; Área de Virosis Emergentes y Zoonóticas (AVEZ), Laboratorio de Ingeniería Genética y Biología Celular y Molecular (LIGBCM), Departamento de Ciencia y Tecnología (DCyT), Universidad Nacional de Quilmes (UNQ), Roque Sáenz Peña 352, B1876BXD, Bernal, Buenos Aires, Argentina. Electronic address: sandra.goni@unq.edu.ar.
Jazyk: angličtina
Zdroj: Protein expression and purification [Protein Expr Purif] 2019 Jan; Vol. 153, pp. 18-25. Date of Electronic Publication: 2018 Aug 17.
DOI: 10.1016/j.pep.2018.08.008
Abstrakt: Saint Louis encephalitis virus (SLEV) and West Nile virus (WNV) are two of the major causes of arboviral encephalitis in the Americas. The co-circulation of related flaviviruses in the Americas and prior vaccination against flaviviruses pose problems to the diagnostic specificity of serological assays due to the development of cross-reactive antibodies. An accurate diagnosis method capable of differentiating these related viruses is needed. NS1 is a glycosylated, nonstructural protein, of about 46 kDa which has a highly conserved structure. Anti-NS1 antibodies can be detected within 4-8 days after the initial exposure and NS1 is the least cross-reactive of the flaviviral antigens. This study was aimed to generate SLEV and WNV NS1 recombinants proteins for the development of a flavivirus diagnostic test. Local Argentinian isolates were used as the source of NS1 gene cloning, expression, and purification. The protein was expressed in Escherichia coli as inclusion bodies and further purified by metal-chelating affinity chromatography (IMAC) under denaturing conditions. Human sera from SLEV and WNV positive cases showed reactivity to the recombinant NS1 proteins by western blot. The unfolded NS1 proteins were also used as immunogens. The polyclonal antibodies elicited in immunized mice recognized the two recombinant proteins with differential reactivity.
(Copyright © 2018 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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