公开(公告)号：US07015023B1

公开(公告)日：2006-03-21

申请号：US10009472

申请日：2000-05-02

申请人： Eric Lam ,  Olga del Pozo

发明人： Eric Lam ,  Olga del Pozo

IPC分类号： C12P21/06 ,  C12N9/00 ,  C12N9/42 ,  C07H21/04

CPC分类号： C12Y302/01031 ,  C07K14/721 ,  C07K2319/00 ,  C12N9/2402 ,  C12Q1/37 ,  C12Q1/6897

摘要： A novel assay system is disclosed for detecting the presence or amount of selected active proteases in biological samples. The assay system utilizes a chimeric protease detector protein composed of three domains: (1) a repressor domain, (2) a protease cleavage domain specific for the protease to be assayed, and a reporter domain. The reporter domain is not detectable when linked to the repressor domain, but becomes detectable upon release from the repressor domain by protease-mediated cleavage. Thus, the activity of the selected protein can be determined by measuring the amount of detectable reporter in the sample. Methods and test kits for using the novel assay system in a variety of in vitro and in vivo applications are also disclosed.

摘要翻译： 公开了一种用于检测生物样品中所选择的活性蛋白酶的存在或量的新型测定系统。 测定系统利用由三个结构域组成的嵌合蛋白酶检测蛋白：（1）阻遏物结构域，（2）对要测定的蛋白酶特异的蛋白酶切割结构域和报道结构域。 当与阻遏物结构域连接时，报道结构域不可检测，但通过蛋白酶介导的切割从抑制子结构域释放时可以检测到。 因此，可以通过测量样品中可检测的报告物的量来确定所选蛋白质的活性。 还公开了在多种体外和体内应用中使用新型测定系统的方法和测试试剂盒。