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Plasma prekallikrein activation and kallikrein production assay

机译:血浆前激肽释放酶激活和激肽释放酶产生测定

摘要

The present invention provides a co-activator for prekallikrein and a prekallikrein-activating method in the absence of FXIIa by using the co-activator which is selected from the group consisting of heat shock protein 90, a phosphate ion and mixtures thereof. The prekallikrein co-activator of the present invention represents a part of an in vitro system for activating prekallikrein, the system comprising said activator as well as a high-molecular weight kininogen and zinc ion. The present invention also comprises a method for activating prekallikrein in the presence of said prekallikrein activator to produce kallikrein, the method being useful in a method for screening the activity of a substance to act as a modulator of the production of kallikrein in the absence of FXIIa. The method of the present invention for activating prekallikrein in the absence of FXIIa comprises activating prekallikrein by mixing prekallikrein with the prekallikrein co-activator, a composition comprising high molecular weight kininogen, and zinc ions. A method for screening the activity of a test substance for the production of kallikrein comprises activating prekallikrein, and determining quantitatively the kallikrein produced in said reaction. In the activity measuring method of the present invention, the activity (a promoting or inhibiting ability) of the tested substance or drug against the production of kallikrein in the absence of FXIIa can be easily, simply, reliably, quickly and precisely measured.
机译:本发明提供了用于前激肽释放酶的共活化剂和在不存在FXIIa的情况下通过使用选自热激蛋白90,磷酸根离子及其混合物的共激活剂的前激肽释放酶的活化方法。本发明的前激肽释放酶共激活剂代表用于激活激肽释放酶的体外系统的一部分,该系统包含所述激活剂以及高分子量激肽原和锌离子。本发明还包括一种在所述前激肽释放酶活化剂存在下活化前激肽释放酶以产生激肽释放酶的方法,该方法可用于筛选在没有FXIIa的情况下用作激肽释放酶产生的调节剂的物质的活性的方法。 。本发明在不存在FXIIa的情况下活化前激肽释放酶的方法包括通过将前激肽释放酶与前激肽释放酶共活化剂,包含高分子量激肽原的组合物和锌离子混合来活化前激肽释放酶。筛选用于产生激肽释放酶的测试物质的活性的方法包括活化激肽释放酶,并定量测定在所述反应中产生的激肽释放酶。在本发明的活性测量方法中,在不存在FXIIa的情况下,可以容易,简单,可靠,快速和精确地测量被测物质或药物对激肽释放酶产生的活性(促进或抑制能力)。

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