组织型纤溶酶原激活剂的发光分析法及其应用

 本文建立了组织型纤溶酶原激活剂(t-PA)活力的发光固相测定方法。用氨基乙基丁基异鲁米诺(ABEI)标记纤维蛋白原(Fg),在一定条件下,t-PA作用于固相(包被ABEI-Fg),产生纤维蛋白的降解产物。测定可溶性降解产物的发光强度,即能计算t-PA活性。该方法的标准曲线范围对t-PA为0.156IU/mL～40IU/mL。灵敏度可达0.156IU/mL。回收率为98.6％(n=27)。批内批间变异系数分别为6.6％及10.3％。该方法曾用于检测细胞培养液中提取t-PA样品及t-PA基因表达时培养液中t-PA的活性。也曾用于检测从组织中纯化t-PA样品及血浆中t-PA活性的测定。文中讨论了该方法与其它方法优缺点的比较。

Luminescence Assay for Determing the Activity of Tissue-type Plasminogen Activator and Its Applications

A luminescence solid phase assay for tissue type plasminogen activator(t-PA) was developed. Amino ethyl butyl isoluminol (ABED was used to label fibrinogen. Under certain Condition ABEI-fibrin degradation products was produced by t-PA which reacted with the sold phase ABEI-fibrin. By detarming the luminescence intensity of ABEI-fibrin degradation products, t-PA activity can be calculated. The range of the standard curve for t-PA was 0.156-40 IU/mL, the sensitivity was about 0.156 IU/mL. The intra and inter assay coefficients of variation were 6.6% and 10.3% respectively. Recovery tests were determined to be 98.6%(n = 27). The adventages and disadvantages of this assay compared with other assays was discussed in this paper.