Pedersen A N, Brünner N, Høyer-Hansen G, Hamer P, Jarosz D, Larsen B, Nielsen H J, Stephens R W
The Finsen Laboratory, Rigshospitalet, Strandboulevarden 49, 2100 Copenhagen, Denmark.
Clin Chem. 1999 Aug;45(8 Pt 1):1206-13.
The complex between urokinase (uPA) and its type-1 inhibitor (PAI-1) is formed exclusively from the active forms of these components; thus, the complex concentration in a biological sample may reflect the ongoing degree of plasminogen activation. Our aim was to establish an ELISA for specific quantification of the uPA:PAI-1 complex in plasma of healthy donors and breast cancer patients.
A kinetic sandwich format immunoassay was developed, validated, and applied to plasma from 19 advanced-stage breast cancer patients, 39 age-matched healthy women, and 31 men.
The assay detection limit was <2 ng/L, and the detection of complex in plasma was validated using immunoabsorption, competition, and recovery tests. Eighteen cancer patients had a measurable complex concentration (median, 68 ng/L; range, <16 to 8700 ng/L), whereas for healthy females and males the median signal values were below the detection limit (median, <16 ng/L; range, <16 to 200 ng/L; P <0.0001). For patient plasma, a comparison with total uPA and PAI-1 showed that the complex represented a variable, minor fraction of the uPA and PAI-1 concentrations of each sample.
The reported ELISA enables detection of the uPA:PAI-1 complex in blood and, therefore, the evaluation of the complex as a prognostic marker in cancer.
尿激酶（uPA）与其1型抑制剂（PAI-1）之间的复合物仅由这些成分的活性形式形成；因此，生物样品中的复合物浓度可能反映纤溶酶原激活的进行程度。我们的目的是建立一种酶联免疫吸附测定法（ELISA），用于特异性定量健康供体和乳腺癌患者血浆中的uPA:PAI-1复合物。
开发、验证了一种动力学夹心式免疫测定法，并将其应用于19例晚期乳腺癌患者、39例年龄匹配的健康女性和31例男性的血浆。
该测定法的检测限<2 ng/L，通过免疫吸附、竞争和回收率测试验证了血浆中复合物的检测。18例癌症患者的复合物浓度可测量（中位数为68 ng/L；范围为<16至8700 ng/L），而健康女性和男性的中位信号值低于检测限（中位数<16 ng/L；范围为<16至200 ng/L；P<0.0001）。对于患者血浆，与总uPA和PAI-1的比较表明，复合物在每个样品的uPA和PAI-1浓度中占可变的小部分。
所报道的ELISA能够检测血液中的uPA:PAI-1复合物，因此可将该复合物评估为癌症的预后标志物。
