A chemiluminescence flow immunosensor based on a porous monolithic metacrylate and polyethylene composite disc modified with Protein G
A chemiluminescence flow immunosensor based on a porous monolithic metacrylate and polyethylene composite disc modified with Protein G
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DOI：
10.1016/j.bios.2003.08.008
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发表时间：
2004-03-15
影响因子：
12.6
通讯作者：
Emnéus, J
中科院分区：
文献类型：
--
作者：
Jain, SR;Borowska, E;Emnéus, J
研制了一种通用、快速、灵敏的新型流动免疫传感器。基础是用蛋白G修饰的单片多孔聚（甲基丙烯酸缩水甘油酯-co-三甲基丙烯酸异丙酯）聚合物圆盘，置于喷泉型流动池隔室中，紧邻光电倍增管（PMT）。分析物和HRP标记的分析物衍生物（示踪剂）竞争抗分析物抗体结合位点。然后将混合物注射到流动免疫传感器系统中，其中所形成的分析物-和示踪物-抗体复合物被整体蛋白G盘捕获。在第二步中，通过注射鲁米诺、对碘苯酚和H2 O2，用辣根过氧化物酶（HRP）产生增强的化学发光（CL），测定与样品中分析物浓度成反比的结合示踪剂的量。第三步也是最后一步是需要再生蛋白G圆盘，以便进行新的分析循环。圆盘免疫传感器系统的性能进行了比较与一个步骤的连续流动注射免疫测定（FIIA）系统，使用相同的试剂和蛋白G柱，从各种水样（微孔，自来水和地表水）的测定灵敏度和基质效应的影响。对于FIIA，PBS和地表水（SW）中分析物阿特拉津的检测限为0.208 +/- 0.004 μ g l（-1）（PBS）和0.59 +/- 0.120 μ g l（-1）（SW），盘式免疫传感器的检测限为0.033 +/- 0.003 μ g l（-1）（PBS）和0.038 +/- 0.003 μ g l（-1）（SW）。两个系统的统计学比较表明，圆盘免疫传感器的结果显着较少的影响，样品基质，这是解释的事实，即在FIIA中的样品与基质同时到达检测器，而这些在圆盘免疫传感器系统中是分开的时间。(C)2003 Elsevier B. V.保留所有权利。
A generic, fast, sensitive and new type of flow immunosensor has been developed. The basis is a monolithic porous poly(glycidyl methacrylate-co-trimethylolpropane trimethacrylate) polymer disc modified with protein G, placed in a fountain type flow cell compartment, in close proximity to a photomultiplier tube (PMT). Analyte and HRP labelled analyte derivative (tracer) compete for anti-analyte antibody binding sites. The mixture is then injected into the flow immunosensor system where the formed analyte- and tracer-antibody complexes are trapped by the monolithic protein G disc. The amount of bound tracer, inversely related to the concentration of analyte in the sample, is determined in a second step by injection of luminol, p-iodophenol and H2O2, generating enhanced chemiluminescence (CL) with horseradish peroxidase (HRP). A third and final step is need for regeneration of the protein G disc so that a new analysis cycle can take place. The performance of the disc immunosensor system was compared with a one step continuous flow injection immunoassay (FIIA) system, using the same reagents and a protein G column, in terms of assay sensitivity and influence of matrix effects from various water samples (millipore-, tap- and surface water). The detection limit for the analyte atrazine in PBS and surface water (SW) was 0.208 +/- 0.004 mug l(-1) (PBS) and 0.59 +/- 0.120 mug l(-1) (SW) for the FIIA and 0.033 +/- 0.003 mug l(-1) (PBS) and 0.038 +/- 0.003 mug l(-1) (SW) for the disc immunosensor. Statistical comparison of the two systems shows that the disc immunosensor results were significantly less influenced by the sample matrix, which is explained by the fact that the sample in the FIIA arrives simultaneously with the matrix to the detector, whereas these are separated in time in the disc immunosensor system. (C) 2003 Elsevier B.V. All rights reserved.