纤溶酶-α_2-抗纤溶酶复合物单克隆抗体的制备及其应用

Preparation and Application of Monoclone Antibodies of Plasmin-α 2-antiplasmin Complexes.

目的:制备纤溶酶-α2-抗纤溶酶复合物（plasmin-α2-antiplasmin complex,PAP）的单克隆抗体并建立其酶免疫检测方法。方法:从人血浆中提纯的PAP作为免疫原制备特异性单克隆抗体,通过双抗体夹心法酶联免疫吸附试验（ELISA）对该单克隆抗体应用价值进行评估。结果:获得特异针对PAP新抗原的单抗3C10和针对PAP及纤溶酶原（Plg）的单抗2B9,效价均为1.25X10-6,其对PAP的亲和常数分别为4.69x10-10M和5.62X10-9M。以此建立的双抗体夹心ELISA检测PAP在0～160ng/ml范围内呈良好的线性关系,批内CV<7％,检测敏感度为5ng/ml,与AD公司的PAP试剂盒相关性良好（r=0.9629）。结论:本法可应用于临床评估纤溶系统的激活状态。

Objective: To prepare the monoclone antibodies(McAbs)of plasmin-α2- antiplasmin complexes (PAP) and use its McAb to establish an ELISA detective method in plasma. Methods: Preparing the purified PAP from fresh human plasma to be used as the monoclonal antibodies, then evaluating the value of application by the method of sandwiched ELISA. Results: Specific McAb 3C10 to PAP, as well as McAb 2B9 to PAP and plasminogen was obtained. Their liter was 1. 25x10-6, and the affinity constant was 4. 69x10-10 M and 5. 62 X 10-9 M respectively. ELISA based on these two McAbs showed a good linear correlation within the range of 0-160ng/ml PAP and intra-assay variation coefficients were less than 7. 0% . The sensitivity of this test was about 5ng/ml and the correlation between ELISA and a commercial PAP kit (ADI) was good (r=0. 9629). Conclusions: This method is suitable for evaluating the activation rate of the fibrinolytic system.