Enzyme-linked coagulation assay: a clot-based, solid-phase assay for thrombin.
Enzyme-linked coagulation assay: a clot-based, solid-phase assay for thrombin.
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酶联凝血测定：基于凝块的凝血酶固相测定。
DOI：
10.1016/0003-2697(85)90310-0
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发表时间：
1985
影响因子：
2.9
通讯作者：
Rothberger,H
中科院分区：
文献类型：
--
作者：
Doellgast,GJ;Rothberger,H
本文提出了一种新的凝血酶固相微量滴定板测定法，以纤维蛋白原结合在微量滴定板的威尔斯孔上，过氧化物酶-纤维蛋白原溶液作为指示体系。当向混合物中加入少量凝血酶时，形成过氧化物酶-纤维蛋白和板结合纤维蛋白，并且过氧化物酶-纤维蛋白结合板结合纤维蛋白。过氧化物酶-纤维蛋白结合的量与凝血酶浓度和孵育时间成比例。使用该测定法，在150 μl样品中测定的凝血酶浓度低至0.25 ng/ml（0.006 nm）。在存在特异性抑制剂苯甲脒和d-苯丙氨酰-L-脯氨酰-L-精氨酸氯甲基酮的情况下，凝血酶活性在与其亲和力和作用机制一致的抑制剂相对浓度下降低。酶联凝血试验通常可用作常规“基于凝块”的凝血试验的高灵敏度和方便的替代方法。
A new, solid-phase microtiter plate assay for thrombin has been developed, using fibrinogen bound to wells of a microtiter plate and peroxidase-fibrinogen in solution as an indicator system. When small amounts of thrombin are added to the mixture, peroxidase-fibrin and plate-bound fibrin are formed, and the peroxidase-fibrin binds to the plate-bound fibrin. The amount of peroxidase-fibrin binding is proportional to the thrombin concentration and time of incubation. Using this assay, thrombin was measured at concentrations as low as 0.25 ng/ml (0.006 nm) in 150 μl of sample. In the presence of the specific inhibitors benzamidine and d-phenylalanyl-l-prolyl-l-arginine chloromethyl ketone, the thrombin activity is reduced, at relative concentrations of inhibitors consistent with their affinities and mechanisms of action. The enzyme-linked coagulation assay is generally useful as a highly sensitive and convenient alternative to conventional “clot-based” tests of coagulation.