METHOD FOR MASS PURIFYING NONSTRUCTURAL PROTEIN 3-4A OF HEPATITIS C VIRUS

摘要

PURPOSE: Provided are methods for mass purifying and activating nonstructural protein 3-4A of hepatitis C virus, so as to develop an inhibitor of nonstructural protein 3-4A of hepatitis C virus, thereby providing nonstructural proteins in a great quantity required for development of an inhibitor of nonstructural protein 3-4A of hepatitis C virus. CONSTITUTION: A method for mass purifying nonstructural protein 3-4A of hepatitis C virus comprises steps of: (a) separating an inactive inclusion body including nonstructural protein 3-4A by crushing and centrifuging E.coli cell; (b) solubilizing the inactive inclusion body with buffer solution including Tris (pH 8.0) of 20mM and urea of 8M; (c) diluting inactive nonstructural protein 3-4A by using buffer solution including urea and refolding this by dialysis with buffer solution, so as to activate this; and (d) separating inactive nonstructural protein 3-4A by using Ni- chelating histidine-affinity gel chromatography and then obtaining this by elution thereof with buffer solution including imidazole.