Measurement of plasminogen activator inhibitor 1 in biologic fluids with a murine monoclonal antibody-based enzyme-linked immunosorbent assay.
Measurement of plasminogen activator inhibitor 1 in biologic fluids with a murine monoclonal antibody-based enzyme-linked immunosorbent assay.
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DOI：
10.1182/blood.v71.1.220.bloodjournal711220
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发表时间：
1988
期刊：
影响因子：
20.3
通讯作者：
P. Declerck;M. Alessi;M. Verstreken;E. Kruithof;I. Juhan-vague;D. Collen
中科院分区：
文献类型：
--
作者：
P. Declerck;M. Alessi;M. Verstreken;E. Kruithof;I. Juhan-vague;D. Collen
基于从HT-1080纤维肉瘤细胞中纯化的两种抗PAI-1的小鼠单克隆抗体，建立了生物体液中纤溶酶原激活物抑制剂-1 （PAI-1）的酶联免疫吸附试验。本试验在血浆中的灵敏度下限为2 ng/mL。与游离PAI-1相比，该检测对PAI-1/人组织型纤溶酶原激活物（t-PA）复合物的敏感性低12倍。测定内变异系数、测定间变异系数和稀释间变异系数分别为5.2%、8.0%和7.1%。健康受试者血小板缺乏血浆PAI-1水平为18 +/- 10 ng/mL（平均+/- SD， n = 45）。在冷冻和解冻后的富血小板血浆中，92%的PAI-1抗原从血小板中释放出来，而在相应的贫血小板血浆中仅发现8%的PAI-1抗原。在健康受试者血小板缺乏的血浆中，PAI活性与PAI-1抗原呈线性相关（r = 0.80）。血浆中大约三分之二的PAI-1抗原具有功能活性，而血小板中释放的PAI-1抗原只有5%具有活性。在怀孕期间，PAI-1抗原水平逐渐增加，达到控制值的三到六倍。复发性深静脉血栓患者血浆PAI-1水平为44 +/- 20 ng/mL（平均+/- SD， n = 7），处于临床沉默期。其中4例高于38 ng/mL（平均正常值+/- 2 SD）。本试验基于稳定和可重复的试剂，可以在生物液体中特异性测定PAI-1抗原。它可以促进实验室间的比较，并有助于进一步研究PAI-1在与纤维蛋白溶解受损和/或血栓形成倾向相关的临床状况中的作用，以及PAI-1在血小板中的作用。
An enzyme-linked immunosorbent assay for plasminogen activator inhibitor-1 (PAI-1) in biologic fluids was developed on the basis of two murine monoclonal antibodies raised against PAI-1 purified from HT-1080 fibrosarcoma cells. The lower limit of sensitivity of the assay in plasma is 2 ng/mL. The assay is 12 times less sensitive toward the PAI-1/human tissue-type plasminogen activator (t-PA) complex as compared with free PAI-1. The intraassay, interassay, and interdilution coefficients of variation are 5.2%, 8.0%, and 7.1%, respectively. The level of PAI-1 in platelet-poor plasma of healthy subjects is 18 +/- 10 ng/mL (mean +/- SD, n = 45). In platelet-rich plasma after freezing and thawing, 92% of PAI-1 antigen is released from platelets, whereas only 8% is found in the corresponding platelet-poor plasma. In platelet-poor plasma from healthy subjects, a linear correlation (r = 0.80) was found between PAI activity and PAI-1 antigen. In plasma approximately two thirds of the PAI-1 antigen was functionally active, whereas only 5% of the PAI-1 antigen released from platelets was active. During pregnancy a progressive increase of PAI-1 antigen levels up to three- to sixfold the control value was observed. In plasma of patients with recurrent deep vein thrombosis, PAI-1 levels were 44 +/- 20 ng/mL (mean +/- SD, n = 7), during a clinically silent phase. Four of these patients had a level above 38 ng/mL (mean +/- 2 SD of normal). The present assay, based on stable and reproducible reagents, allows the specific determination of PAI-1 antigen in biologic fluids. It may facilitate interlaboratory comparisons and be useful for further investigations of the role of PAI-1 in clinical conditions associated with impaired fibrinolysis and/or a tendency to thrombosis and investigations of the role of PAI-1 in platelets.