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。
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
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2004年第6期381-384,共4页
Chinese Journal of Laboratory Medicine
基金
上海市科技发展基金资助项目 (9941 1 90 0 3)
