國家衛生研究院 NHRI:Item 3990099045/1161

NHRI > Institute of Biotechnology and Pharmaceutical Research > John Tsu-An Hsu > Periodical Articles > Item 3990099045/1161

Please use this identifier to cite or link to this item: http://ir.nhri.org.tw/handle/3990099045/1161

Title:	High-throughput cell-based screening for hepatitis C virus NS3/4A protease inhibitors
Authors:	Lee, JC;Yu, MC;Lien, TW;Chang, CF;Hsu, JTA
Contributors:	Division of Biotechnology and Pharmaceutical Research
Abstract:	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.
Keywords:	Biochemical Research Methods;Pharmacology & Pharmacy
Date:	2005-08
Relation:	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
Appears in Collections:	[John Tsu-An Hsu] Periodical Articles

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