A new easy method for specific measurement of active myeloperoxidase in human biological fluids and tissue extracts
A new easy method for specific measurement of active myeloperoxidase in human biological fluids and tissue extracts
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DOI：
10.1016/j.talanta.2009.07.052
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发表时间：
2009-12-15
期刊：
影响因子：
6.1
通讯作者：
Serteyn, D.
中科院分区：
文献类型：
--
作者：
Franck, T.;Kohnen, S.;Serteyn, D.
SIEFED（“特异性免疫提取后进行酶检测”）方法已经开发用于特异性检测马髓过氧化物酶（MPO）的活性，该方法适用于特异性测量生物体液或组织提取物中的活性人 MPO。该方法包括通过固定化抗 MPO 抗体从水溶液中提取 MPO，然后进行洗涤（以消除提取介质和生物液体及其可能的干扰分子），并使用含有荧光底物、H2O2 和亚硝酸根离子作为反应增强剂的检测系统测量 MPO 的活性。 SIEFED 用于研究人体生物体液（血浆、支气管肺泡灌洗液和颈动脉提取物上清液）中的活性 MPO。人 MPO 的 SIEFED 的灵敏度限值为 0.080 mU/mL，在 0.156 至 473 mU/mL 的广泛 MPO 活性范围内显示出良好的精度，测定内和测定间变异系数分别低于 10% 和 20%。人 MPO 的 SIEFED 将有助于特异性检测复杂液体中的活性 MPO，并且可以与 ELISA 互补，以确定健康中的活性/总 MPO 比率。志愿者和患者，尤其是患有慢性或急性炎症性疾病的患者。 (C) 2009 Elsevier B.V. 保留所有权利。
The SIEFED ("Specific Immunological Extraction Followed by Enzymatic Detection") method already developed for the specific detection of the activity of equine myeloperoxidase (MPO) was adapted for the specific measurement of active human MPO in biological fluids or tissue extracts. The method consists of the extraction of MPO from aqueous solutions by immobilized anti-MPO antibodies followed by a washing(to eliminate the extraction medium and the biological fluid with their possible interfering molecules) and the measurement of the activity of MPO with a detection system containing a fluorogenic substrate, H2O2 and nitrite ions as reaction enhancer. The SIEFED was applied to study active MPO in human biological fluids (plasma, bronchoalveolar lavage fluid and supernatant from carotids extracts). The SIEFED for human MPO has a sensitivity limit of 0.080 mU/mL and showed good precision with intra- and inter-assay coefficients of variation below 10 and 20% respectively within a broad range of MPO activities establish from 0.156 to 473 mU/mL The SIEFED for human MPO will be useful for the specific detection of active MPO in complex fluids and can be complementary to an ELISA to determine an active/total MPO ratio in healthy volunteers and patients especially in case of chronic or acute inflammatory diseases. (C) 2009 Elsevier B.V. All rights reserved.