Disposable amperometric immunosensor for the detection of polycyclic aromatic hydrocarbons (PAHs) using screen-printed electrodes
Disposable amperometric immunosensor for the detection of polycyclic aromatic hydrocarbons (PAHs) using screen-printed electrodes
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DOI：
10.1016/s0956-5663(02)00112-4
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发表时间：
2003-01-01
影响因子：
12.6
通讯作者：
Guilbault, GG
中科院分区：
文献类型：
--
作者：
Fähnrich, KA;Pravda, M;Guilbault, GG
研制了一种多环芳烃(PAHs)电流型免疫传感器。该免疫传感器是基于一次性丝网印刷碳电极。包被抗原为菲-9-甲醛通过己二酸二肼偶联牛血清白蛋白(BSA)。抗体为鼠抗菲单抗。碱性磷酸酶(AP)和底物对氨基苯基磷酸(PAPP)在+300 mV(vs.Ag/AgCl)下检测。对不同的检测方法进行比较。间接共暴露竞争法检测菲的最低检出限为0.8 ng/ml(800 Ppt)，IC50为7.1 ng/ml(7.1 ppb)。以8.8mug/ml菲-牛血清白蛋白为底物，间接竞争法检测菲的最低检出限为2 ng/ml(IC50：15 ng/ml)，间接置换法的最低检出限为2 ng/ml(IC50：11 ng/ml)。当包被浓度为2.2µg/ml时，间接竞争法检测的最低检出限为0.25 ng/ml(250 Ppt)。研究了涂层浓度对传感器性能的影响。对16种重要的多环芳烃进行了交叉反应测试。菲和大黄烯有很强的交叉反应，而苯并[g，h，i]茂和二苯并[a，h]茂没有交叉反应。(C)2002 Elsevier Science B.V.保留所有权利。
An amperometric immunosensor for polycyclic aromatic hydrocarbons (PAHs) was developed. The immunosensor was based on disposable screen-printed carbon electrodes. The coating antigen used was phenanthrene-9-carboxaldehyde coupled to bovine serum albumin (BSA) via adipic acid dihydrazide. Antibodies were monoclonal mouse anti-phenanthrene. The enzyme alkaline phosphatase (AP) was used in combination with the substrate p-aminophenyl phosphate (pAPP) for detection at +300 mV (vs. Ag/AgCl). Various assay types were compared. Good results were achieved with an indirect co-exposure competition assay with a LOD of 0.8 ng/ml (800 ppt) and an IC50 of 7.1 ng/ml (7.1 ppb) for phenanthrene. An indirect competition assay could detect phenanthrene with a LOD of 2 ng/ml (IC50: 15 ng/ml) and an indirect displacement assay with a LOD of 2 ng/ml (IC50: 11 ng/ml) at a 5 mul surface coating of 8.8 mug/ml phenanthrene-BSA conjugate. A coating concentration of 2.2 mug/ml allowed detection with a LOD of 0.25 ng/ml (250 ppt) with the indirect competition assay. The influence of the coating concentration on the sensor performance was investigated. Cross-reactivities were tested for 16 important PAHs. Anthracene and chrysene showed strong cross-reactivity, whereas benzo[g,h,i]perylene and dibenzo[a,h]anthracene showed no cross-reactivity. (C) 2002 Elsevier Science B.V. All rights reserved.