Protein refolding based on high hydrostatic pressure and alkaline pH
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2019
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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.
Como referenciar
CHURA-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. PLoS One, v. 14, n. 1, p. 1-14, 2019. DOI: 10.1371/journal.pone.0211162. Disponível em: http://repositorio.ipen.br/handle/123456789/30068. Acesso em: 28 Mar 2025.
Esta referência é gerada automaticamente de acordo com as normas do estilo IPEN/SP (ABNT NBR 6023) e recomenda-se uma verificação final e ajustes caso necessário.