Cell-based assay systems for examining hcv ns3 protease activity

摘要

Assays for identifying levels of hepatitis C virus NS3 protease activity are provided herein, and involve the transfection into mammalian cells of cDNA corresponding to HCV nucleic acid sequences encoding both the viral NS3 protease activity and an NS3 polyprotein precursor substrate including an NS3 cleavage site. The presence of mature, processed substrate in lysates obtained from the transfected cells is indicative of the level of activity of the NS3 protease in the cells. These assays can thus also be used as a screen to examine the activity of a particular compound in inhibiting NS3 protease activity, by conducting the assays in the absence and presence of the compound. The degree to which NS3 protease activity is expressed in the former assays over the latter is indicative of the compound's activity in inhibiting the NS3 protein substrate cleavage, and thus also, of inhibiting HCV replication.