| dc.creator | CHURA-CHAMBI, ROSA M. | |
| dc.creator | SILVA, CLEIDE M.R. da | |
| dc.creator | PEREIRA, LENNON R. | |
| dc.creator | BARTOLINI, PAOLO | |
| dc.creator | FERREIRA, LUIS C. de S. | |
| dc.creator | MORGANTI, LIGIA | |
| dc.date | 2019 | |
| dc.date | 2019-08-09T14:50:42Z | |
| dc.date | 2019-08-09T14:50:42Z | |
| dc.date.accessioned | 2023-09-28T14:11:29Z | |
| dc.date.available | 2023-09-28T14:11:29Z | |
| dc.identifier | 1932-6203 | |
| dc.identifier | http://repositorio.ipen.br/handle/123456789/30068 | |
| dc.identifier | 1 | |
| dc.identifier | 14 | |
| dc.identifier | 10.1371/journal.pone.0211162 | |
| dc.identifier | 62.676 | |
| dc.identifier | 91.00 | |
| dc.identifier.uri | https://repositorioslatinoamericanos.uchile.cl/handle/2250/9000306 | |
| dc.description | In 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. | |
| dc.description | Funda????o de Amparo ?? Pesquisa do Estado de S??o Paulo (FAPESP) | |
| dc.description | Conselho Nacional de Desenvolvimento Cient??fico e Tecnol??gico (CNPq) | |
| dc.description | FAPESP: 15/02574-0 | |
| dc.description | CNPq: 130256/2015-3 | |
| dc.format | 1-14 | |
| dc.relation | PLoS One | |
| dc.rights | openAccess | |
| dc.subject | viruses | |
| dc.subject | viral diseases | |
| dc.subject | proteins | |
| dc.subject | ph value | |
| dc.subject | hydrostatics | |
| dc.subject | pressure dependence | |
| dc.subject | immunoassay | |
| dc.title | Protein refolding based on high hydrostatic pressure and alkaline pH | |
| dc.type | Artigo de peri??dico | |
| dc.coverage | I | |
