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1. 发明申请

US20070172936A1 Purified active HCV NS2/3 protease 审中-公开
公开(公告)号：US20070172936A1
公开(公告)日：2007-07-26
申请号：US11223638
申请日：2005-09-09
申请人： Diane Thibeault , Daniel Lamarre , Roger Maurice , Louise Pilote , Armin Pause
发明人： Diane Thibeault , Daniel Lamarre , Roger Maurice , Louise Pilote , Armin Pause
IPC分类号： C12N9/99 , C12N9/48 , C07K7/08
CPC分类号： C07K14/005 , C12N9/506 , C12N2770/24222 , C12Q1/37 , G01N33/6893 , G01N2333/18 , G01N2500/00 , G01N2500/04
摘要： A method for producing a refolded, inactive form of recombinantly produced NS2/3 protease which comprises the steps of: a) purifying the protease from inclusion bodies in the presence of a chaotropic agent; and b) refolding the purified protease by contacting it with a reducing agent and lauryldiethylamine oxide (LDAO) in the presence of reduced concentration of chaotropic agent or polar additive. The invention further comprises a method for activating this refolded inactive NS2/3 protease by adding an activation detergent. This method produces large amounts of the active NS2/3 protease to allow small molecules and ligands to be screened as potential inhibitors of NS2/3 protease, which may be useful as therapeutic agents against HCV.

2. 发明授权

US06815159B2 Purified active HCV NS2/3 protease 有权
标题翻译：纯化的活性HCV NS2 / 3蛋白酶
公开(公告)号：US06815159B2
公开(公告)日：2004-11-09
申请号：US10017736
申请日：2001-12-14
申请人： Diane Thibeault , Daniel Lamarre , Roger Maurice , Louise Pilote , Armin Pause
发明人： Diane Thibeault , Daniel Lamarre , Roger Maurice , Louise Pilote , Armin Pause
IPC分类号： C12Q170
CPC分类号： C07K14/005 , C12N9/506 , C12N2770/24222 , C12Q1/37 , G01N33/6893 , G01N2333/18 , G01N2500/00 , G01N2500/04
摘要： A method for producing a refolded, inactive form of recombinantly produced NS2/3 protease which comprises the steps of: a) purifying the protease from inclusion bodies in the presence of a chaotropic agent; and b) refolding the purified protease by contacting it with a reducing agent and lauryldiethylamine oxide (LDAO) in the presence of reduced concentration of chaotropic agent or polar additive. The invention further comprises a method for activating this refolded inactive NS2/3 protease by adding an activation detergent. This method produces large amounts of the active NS2/3 protease to allow small molecules and ligands to be screened as potential inhibitors of NS2/3 protease, which may be useful as therapeutic agents against HCV.

3. 发明授权

US07655394B2 Hepatitis C virus NS2/3 activity assay 有权
标题翻译：丙型肝炎病毒NS2 / 3活性测定
公开(公告)号：US07655394B2
公开(公告)日：2010-02-02
申请号：US11553531
申请日：2006-10-27
申请人： Roger Maurice , Diane Thibeault
发明人： Roger Maurice , Diane Thibeault
IPC分类号： C12Q1/00
CPC分类号： C12Q1/37 , G01N2500/02
摘要： The invention provides a novel assay for NS2/3 cleavage detection, particularly, a novel assay for the detection of the NS3 cleavage product in the presence of uncleaved NS2/3 based on discrimination of the activity of the resulting NS3 protease product from the NS3 protease activity of the NS2/3 uncleaved protein. Certain reaction conditions allow this discrimination in situ without having to resort to physical separation of both proteases by decreasing the NS3 protease activity of the NS2/3 protein and increasing the NS3 protease activity of the resulting NS3 protease product, thereby producing a “signal window” upon which it is feasible to distinguish and measure NS3 protease activity produced from the cleavage of NS2/3.
摘要翻译：本发明提供了一种用于NS2 / 3切割检测的新型测定法，特别是基于鉴别来自NS3蛋白酶的所得NS3蛋白酶产物的活性，在未切割的NS2 / 3存在下检测NS3切割产物的新型测定法 NS2 / 3未切割蛋白的活性。某些反应条件允许这种现象的鉴别，而不必通过降低NS2 / 3蛋白的NS3蛋白酶活性并增加所得NS3蛋白酶产物的NS3蛋白酶活性而使两种蛋白酶物理分离，从而产生“信号窗” 从而可以区分和测量NS2 / 3裂解产生的NS3蛋白酶活性。

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