A method for producing a refolded, inactive form of recombinantly produced
NS2/3 protease which comprises the steps of: a) purifying the protease
from inclusion bodies in the presence of a chaotropic agent; and b)
refolding the purified protease by contacting it with a reducing agent and
lauryldiethylamine oxide (LDAO) in the presence of reduced concentration
of chaotropic agent or polar additive. The invention further comprises a
method for activating this refolded inactive NS2/3 protease by adding an
activation detergent. This method produces large amounts of the active
NS2/3 protease to allow small molecules and ligands to be screened as
potential inhibitors of NS2/3 protease, which may be useful as therapeutic
agents against HCV.
Um método para produzir a refolded, o formulário inativo do protease NS2/3 recombinantly produzido de que compreende as etapas: a) purifying o protease dos corpos do inclusion na presença de um agente chaotropic; e b) refolding o protease purified contatando o com um agente reduzindo-se e um óxido do lauryldiethylamine (LDAO) na presença da concentração reduzida do agente chaotropic ou do aditivo polar. A invenção mais adicional compreende um método para ativar isto refolded o protease NS2/3 inativo adicionando um detergente da ativação. Este método produz quantidades grandes do protease NS2/3 ativo para permitir moléculas pequenas e os ligands a ser selecionados como os inibidores potenciais do protease NS2/3, que podem ser úteis como agentes therapeutic de encontro a HCV.
