Association of high pressure and alkaline condition for solubilization of inclusion bodies and refolding of the NS1 protein from zika virus
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BMC Biotechnology
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Background: Proteins in inclusion bodies (IBs) present native-like secondary structures. However, chaotropic agents at
denaturing concentrations, which are widely used for IB solubilization and subsequent refolding, unfold these
secondary structures. Removal of the chaotropes frequently causes reaggregation and poor recovery of bioactive
proteins. High hydrostatic pressure (HHP) and alkaline pH are two conditions that, in the presence of low level of
chaotropes, have been described as non-denaturing solubilization agents. In the present study we evaluated the
strategy of combination of HHP and alkaline pH on the solubilization of IB using as a model an antigenic form of the
zika virus (ZIKV) non-structural 1 (NS1) protein.
Results: Pressure-treatment (2.4 kbar) of NS1-IBs at a pH of 11.0 induced a low degree of NS1 unfolding and led to
solubilization of the IBs, mainly into monomers. After dialysis at pH 8.5, NS1 was refolded and formed soluble
oligomers. High (up to 68 mg/liter) NS1 concentrations were obtained by solubilization of NS1-IBs at pH 11 in the
presence of arginine (Arg) with a final yield of approximately 80% of total protein content. The process proved to be
efficient, quick and did not require further purification steps. Refolded NS1 preserved biological features regarding
reactivity with antigen-specific antibodies, including sera of ZIKV-infected patients. The method resulted in an increase
of approximately 30-fold over conventional IB solubilization-refolding methods.
Conclusions: The present results represent an innovative non-denaturing protein refolding process by means of the
concomitant use of HHP and alkaline pH. Application of the reported method allowed the recovery of ZIKV NS1 at a
condition that maintained the antigenic properties of the protein.
Como referenciar
SILVA, CLEIDE M.R. da; CHURA-CHAMBI, ROSA M.; PEREIRA, LENNON R.; CORDEIRO, YRAIMA; FERREIRA, LUIS C. de S.; MORGANTI, LIGIA. Association of high pressure and alkaline condition for solubilization of inclusion bodies and refolding of the NS1 protein from zika virus. BMC Biotechnology, v. 18, n. 78, 2018. DOI: 10.1186/s12896-018-0486-2. Disponível em: http://repositorio.ipen.br/handle/123456789/29402. Acesso em: 30 Dec 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.