| 摘要 |
PURPOSE: A purification method of protease from hepatitis C virus (HCV) is provided which results high yields and purity of purified protease and saves times for the process of purification. CONSTITUTION: Supernatant of disintegrated E. coli cells expressing nonstructural protein 3 of Korean type HCV is applied to CM-Sepharose column chromatography. Protein is eluted with NaCl gradient. Eluted protein is pooled and fractionated with 30-50% of ammonium sulfate. Fractionated protein precipitate is dissolved in 1M ammonium sulfate, 20mM Tris, 20% of glycerol, 1mM DTT, and 0.1% of beta OG solution and applied to phenyl-Sepharose column and eluted with ammonium sulfate gradient. The final yield of protease is 4mg/ml and purity of protease is 97% based on size exclusion chromatography of HPLC. The improved purification method is more efficient than another purification method. |
