Establishment of a high throughput screening assay for identification of HCV NS3 serine protease inhibitors

• Main Authors: Chieh Yang, 楊婕
• Other Authors: Ju-Chien Cheng
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/5dp9hk

碩士 === 中國醫藥大學 === 醫學檢驗生物技術學系碩士班 === 102 === The prevalence of hepatitis C virus (HCV) infection is all around the word. Until now, over 170 million people were infected by HCV. The virus causes liver diseases such as chronic hepatitis, cirrhosis, and liver cancer. The standard therapy of HCV is pegylated-interferon (Peg-IFN) combined with ribavirin (RBV), but only 50% sustained viral response for HCV genotype 1(GT1). Boceprevir and Telaprevir, which are two linear ketoamide compounds, which covalently bind to the serine protease active-site of HCV NS3, have been approved by FDA to treat HCV infection in 2011. In clinical, these two inhibitors induce HCV genotype 1 virus NS3 protease mutation. The aim of the study is to establish a HCV GT1 NS3/4A protease antiviral screening assay. The wild-type HCV genotype 1 NS3 protease and common resistance sites in clinical of the NS3 protease had been constructed and the expressions were determined by Western blotting. The activities of wild type and mutant NS3 protease were determined by Western blotting and high throughput cell–based luciferase assay. The high throughput drug selection system will be applied to the NS3 proteinase inhibitors that exclude viral escape through common resistance sites in clinic.