Koppert P W, Hoegee-de Nobel E, Nieuwenhuizen W
Gaubius Institute TNO, Leiden, The Netherlands.
Thromb Haemost. 1988 Apr 8;59(2):310-5.
We have developed a sandwich-type enzyme immunoassay (EIA) for the quantitation of fibrin degradation products (FbDP) in plasma with a time-to-result of only 45 minutes. The assay is based on the combination of the specificities of two monoclonal antibodies (FDP-14 and DD-13), developed in our institute. FDP-14, the capture antibody, binds both fibrinogen degradation products (FbgDP) and FbDP, but does not react with the parent fibrin(ogen) molecules. It has its epitope in the E-domain of the fibrinogen molecule on the B beta-chain between amino acids 54-118. Antibody DD-13 was raised using D-dimer as antigen and is used as a tagging antibody, conjugated with horse-radish peroxidase. A strong positive reaction is obtained with a whole blood clot lysate (lysis induced by tissue-type plasminogen activator) which is used as a standard. The EIA does virtually not detect FbgDP i.e. purified fragments X, Y, or FbgDP generated in vitro in plasma by streptokinase treatment. This indicates that the assay is specific for fibrin degradation products. We have successfully applied this assay to the plasma of patients with a variety of diseased states. In combination with the assay previously developed by us for FbgDP and for the total amount of FbgDP + FbDP (TDP) in plasma, we are now able to study the composition of TDP in patients plasma in terms of FbgDP and FbDP.
我们开发了一种夹心型酶免疫测定法（EIA），用于定量血浆中的纤维蛋白降解产物（FbDP），出结果时间仅为45分钟。该测定法基于我们研究所开发的两种单克隆抗体（FDP - 14和DD - 13）的特异性组合。捕获抗体FDP - 14可结合纤维蛋白原降解产物（FbgDP）和FbDP，但不与纤维蛋白原分子反应。它的表位位于Bβ链上纤维蛋白原分子E结构域中氨基酸54 - 118之间。以D - 二聚体为抗原制备的抗体DD - 13用作标记抗体，与辣根过氧化物酶结合。以全血凝块裂解物（由组织型纤溶酶原激活剂诱导裂解）作为标准品可获得强阳性反应。该EIA实际上检测不到FbgDP，即体外经链激酶处理血浆产生的纯化片段X、Y或FbgDP。这表明该测定法对纤维蛋白降解产物具有特异性。我们已成功将该测定法应用于患有各种疾病状态患者的血浆。结合我们之前开发的用于血浆中FbgDP以及FbgDP + FbDP总量（TDP）的测定法，我们现在能够从FbgDP和FbDP的角度研究患者血浆中TDP的组成。
