CHURA-CHAMBI, ROSA M.SILVA, CLEIDE M.R. daPEREIRA, LENNON R.BARTOLINI, PAOLOFERREIRA, LUIS C. de S.MORGANTI, LIGIA2019-08-092019-08-092019CHURA-CHAMBI, ROSA M.; SILVA, CLEIDE M.R. da; PEREIRA, LENNON R.; BARTOLINI, PAOLO; FERREIRA, LUIS C. de S.; MORGANTI, LIGIA. Protein refolding based on high hydrostatic pressure and alkaline pH: application on a recombinant dengue virus NS1 protein. <b>PLoS One</b>, v. 14, n. 1, p. 1-14, 2019. DOI: <a href="https://dx.doi.org/10.1371/journal.pone.0211162">10.1371/journal.pone.0211162</a>. Disponível em: http://repositorio.ipen.br/handle/123456789/30068.1932-6203http://repositorio.ipen.br/handle/123456789/30068In this study we evaluated the association of high hydrostatic pressure (HHP) and alkaline pH as a minimally denaturing condition for the solubilization of inclusion bodies (IBs) generated by recombinant proteins expressed by Escherichia coli strains. The method was successfully applied to a recombinant form of the dengue virus (DENV) non-structural protein 1 (NS1). The minimal pH for IBs solubilization at 1 bar was 12 while a pH of 10 was sufficient for solubilization at HHP: 2.4 kbar for 90 min and 0.4 kbar for 14 h 30 min. An optimal refolding condition was achieved by compression of IBs at HHP and pH 10.5 in the presence of arginine, oxidized and reduced glutathiones, providing much higher yields (up to 8-fold) than association of HHP and GdnHCl via an established protocol. The refolded NS1, 109 ± 9.5 mg/L bacterial culture was recovered mainly as monomer and dimer, corresponding up to 90% of the total protein and remaining immunologically active. The proposed conditions represent an alternative for the refolding of immunologically active recombinant proteins expressed as IBs.1-14openAccessvirusesviral diseasesproteinsph valuehydrostaticspressure dependenceimmunoassayArtigo de periódico11410.1371/journal.pone.0211162https://orcid.org/0000-0002-7870-179362.67691.00