國家衛生研究院 NHRI:Item 3990099045/1161
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    题名: High-throughput cell-based screening for hepatitis C virus NS3/4A protease inhibitors
    作者: Lee, JC;Yu, MC;Lien, TW;Chang, CF;Hsu, JTA
    贡献者: Division of Biotechnology and Pharmaceutical Research
    摘要: Hepatitis C virus ( HCV) encodes a viral protease, nonstructural (NS) 3/4A, that is critical for virus maturation. Although NS3/4A has emerged as a promising target for anti-HCV drug discovery, no anti-HCV therapy has succeeded yet based on inhibition of NS3/4A. We have previously shown that EG(Delta 4AB) SEAP, a reporter consisting of enhanced green fluorescent protein ( EG), the NS3-NS4A protease decapeptide recognition sequence (Delta 4AB), and secreted alkaline phosphatase ( SEAP), is an efficient reporter for reflecting NS3/4A proteolytic activity inside cells. In this study, we describe the generation and characterization of a stable cell line, 293E-EG( Delta 4AB) SEAP-NS3/4A, which constitutively expresses EG(Delta 4AB) SEAP reporter protein and NS3/4A protease. The reporter assay is validated with the compound BILN 2061, a specific and potent peptidomimetic inhibitor of the HCV NS3 protease. Additionally, we show here that this cell line allows screening for NS3/4A protease activity of living cells in 96-well plate format, with a Z factor >0.6. Thus, this cell-based assay may be used for high-throughput screening of chemical libraries.
    关键词: Biochemical Research Methods;Pharmacology & Pharmacy
    日期: 2005-08
    關聯: ASSAY and Drug Development Technologies. 2005 Aug;3(4):385-392.
    Link to: http://dx.doi.org/10.1089/adt.2005.3.385
    JIF/Ranking 2023: http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=NHRI&SrcApp=NHRI_IR&KeyISSN=1540-658X&DestApp=IC2JCR
    Cited Times(WOS): https://www.webofscience.com/wos/woscc/full-record/WOS:000232229000004
    Cited Times(Scopus): http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=27144467123
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