【摘要】 目的　建立纤溶酶 α2 抗纤溶酶复合物 (PAP)的检测方法。方法　从血浆中纯化PAP作为免疫原制备单克隆抗体 (单抗 ) ,建立双抗体夹心法酶联免疫吸附试验 (ELISA) ,并对该法进行评价。结果　获取了特异针对PAP新抗原的单抗LW 3C10 ,和针对PAP及纤溶酶原 (Plg)的单抗LW 2B9。对PAP的亲和常数分别为 4 6 9× 10 10 mol/L、5 6 2× 10 9mol/L。以它们建立的夹心ELISA检测PAP在 0～ 15 0 μg/L范围内线性良好 ,批内、批间CV分别为 4 0 %～ 5 2 %、11 2 %～ 13 6 % ,回收率为 85%～ 10 5 % ,加入α2 抗纤溶酶 (α2 AP)及纤溶酶 α2 巨球蛋白复合物 (P α2 M)不干扰测定 ,与美国ADI公司PAP试剂盒相关良好 (r=0 96 2 9)。急性髓细胞白血病组、急性心肌梗死组、肝病组、健康老年人组的血浆PAP水平显著高于正常对照组 (P <0 0 0 1)。结论　该法可用于评估纤溶系统激活。更多还原

【Abstract】 Objective To establish a method to determine plasmin α 2 antiplasmin complexes (PAP) in plasma Methods Developing and evaluating a sandwiched enzyme linked immunosorbent assay (ELISA) using monoclonal antibodies (MoAbs) raised by immunizing BALB/c mice with PAP purified from fresh human plasma Results MoAb LW3C10 recognized PAP exclusively (K d PAP = 4 69×10 10 mol/L), while MoAbLW2B9 recognized both PAP and plasminogen (K d PAP = 5 62×10 9 mol/L ) ELISA based on these two MoAbs exhibited a good linear correlation within the range of 0 ～ 150 μg/L PAP Under these conditions, recovery of PAP added to normal plasma ranged from 85 % to 105 % Addition of α 2 antiplasmin and plasmin α 2 macroglobulin complexes did not impair detection of PAP Intra assay and inter assay variation coefficients were 4 0 % ～ 5 2 % and 11 2 % ～ 13 6 %, respectively The correlation degree between the ELISA and a commercial kit was highly significant ( r =0 962 9) As compared to controls, higher PAP concentrations ( P < 0 001) were observed in acute myelogenous leukemia, acute myocardial infarction, hepatopathy and elderly subjects Conclusion The described method is suitable in evaluating the activation rate of fibrinolytic system更多还原