Assay of Human Tissue-Type Plasminogen Activator (t-PA) with an Enzyme-Linked Immunosorbent Assay (ELISA) Based on Three Murine Monoclonal Antibodies to t-PA
Assay of Human Tissue-Type Plasminogen Activator (t-PA) with an Enzyme-Linked Immunosorbent Assay (ELISA) Based on Three Murine Monoclonal Antibodies to t-PA
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DOI：
10.1055/s-0038-1660097
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发表时间：
1985-10
影响因子：
6.7
通讯作者：
P. Holvoet;Hugo Cleemput;D. Collen
中科院分区：
文献类型：
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作者：
P. Holvoet;Hugo Cleemput;D. Collen
建立了检测人组织型纤溶酶原激活物(t-PA)的酶联免疫吸附试验(EL ISA)。用两种单抗的混合物包被微滴板，并用与过氧化物酶相连的第三种单抗对t-PA进行定量。该方法的灵敏度下限为0.2 ng/ml，人血中t-PA浓度为3.4±0.8 ng/ml，重复性好，批内、批间和稀释间变异系数分别为3.8%、6.5%和4.9%。T-PA溶栓治疗患者血浆标本的酶联免疫吸附试验结果在较宽的浓度范围内与先前报道的双部位免疫放射分析结果有很好的相关性(r=0.96)。用单抗建立了一套稳定、重复性好的检测体液中t-PA抗原的试剂盒，避免了放射性同位素和多克隆抗体的缺点。
Summary An enzyme-linked immunosorbent assay (ELISA) for the measurement of human tissue-type plasminogen activator (t-PA) was developed. Microtiter plates were coated with a mixture of two monoclonal antibodies and bound t-PA was quantitated with a third monoclonal antibody linked to peroxidase. The lower limit of sensitivity of the assay was 0.2 ng of t-PA per ml. The concentration of antigen in citrated plasma of human subjects was found to be 3.4 ± 0.8 ng/ml. The assay had a good reproducibility with values of 3.8, 6.5 and 4.9 percent respectively for the intra-, inter-assay and inter-dilution variation coefficients. The results of the ELISA assay on plasma samples from patients during thrombolytic therapy with t-PA correlated very well, over a wide concentration range, with those obtained with a previously described two-site immuno-radiometric assay (r = 0.96). This ELISA with monoclonal antibodies constitutes a stable and reproducible set of reagents for the measurement of t-PA antigen in biological fluids, avoiding the disadvantages of the use of radioisotopes and of polyclonal antibodies.