血浆中组织型纤溶酶原激活物及其“快速”抑制剂的测定。

Determination of tissue plasminogen activator and its "fast" inhibitor in plasma.

作者信息

Chmielewska J, Wiman B

出版信息

Clin Chem. 1986 Mar;32(3):482-5.

Abstract

We describe efficient, accurate methods for specific determination of tissue plasminogen activator (t-PA, EC 3.4.21.31) and its "fast" inhibitor in plasma. In this coupled assay, a sample containing t-PA is incubated with plasminogen, a plasmin (EC 3.4.21.7) substrate of low Km and high Kcat, and fibrin as a stimulator. The inhibitor of t-PA is determined by incubating the sample with a known amount of t-PA in excess, then determining the residual t-PA. Both t-PA and t-PA inhibitor can be determined in many samples simultaneously within a few hours. These assays are modifications of procedures described by us (Clin Chim Acta 1983;127:279-88 and Thromb Res 1983;31:427-36). Their accuracy as assessed by analytical recovery of pure t-PA added to blood samples (91 +/- 4%) or of partly purified inhibitor added to plasma samples (102 +/- 10%) is satisfactory, as is their precision. For the t-PA assay the CV was 1.6% (within run) or 4.1% (between run). The corresponding values for the inhibitor assay were 4.5% (within run) or 8.4% (between run) if the inhibitor concentration exceeded 3 arb. units/mL.

摘要

我们描述了高效、准确地特异性测定血浆中组织纤溶酶原激活物（t-PA，EC 3.4.21.31）及其“快速”抑制剂的方法。在这种偶联测定中，将含有t-PA的样品与纤溶酶原、一种低Km和高Kcat的纤溶酶（EC 3.4.21.7）底物以及作为刺激物的纤维蛋白一起孵育。通过将样品与已知过量的t-PA一起孵育，然后测定残留的t-PA来确定t-PA的抑制剂。t-PA和t-PA抑制剂均可在数小时内同时在多个样品中进行测定。这些测定方法是我们之前描述的方法的改进（《临床化学学报》1983年；127：279 - 288以及《血栓研究》1983年；31：427 - 36）。通过向血样中添加纯t-PA的分析回收率（91±4%）或向血浆样品中添加部分纯化的抑制剂的分析回收率（102±10%）评估，其准确性令人满意，精密度也是如此。对于t-PA测定，批内变异系数为1.6%，批间变异系数为4.1%。如果抑制剂浓度超过3 arb.单位/mL，抑制剂测定的相应值批内为4.5%，批间为8.4%。