Viability of allergy (IgE) detection using an alternative aptamer receptor and electrochemical means
Viability of allergy (IgE) detection using an alternative aptamer receptor and electrochemical means
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DOI：
10.1016/j.snb.2006.09.024
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发表时间：
2007-01-30
影响因子：
8.4
通讯作者：
Guilbault, George G.
中科院分区：
文献类型：
--
作者：
Papamichael, Katerina I.;Kreuzer, Mark P.;Guilbault, George G.
目前的工作描述了一个一次性的电化学传感器的基础上的适体受体（aptasensor）的系统开发。适体执行的最佳条件已通过酶联寡核苷酸测定（ELISA）进行了彻底研究，并进行了操作以在最终传感器设置中实现足够的灵敏度。研究的参数包括缓冲液类型、试剂浓度分布、pH、离子效应和热稳定性。然后，方法转移允许利用差分脉冲伏安法作为转导模式制备适体传感器。所选的主要系统用于检测过敏（IgE），过敏是由于暴露于食物和/或环境中的多种过敏原而存在于人体中的。该系统采用生物素化形式的适体，采用竞争性形式进行IgE检测。酶的作用是通过随后使用的抗生物素蛋白-碱性磷酸酶标记比色或电化学检测完成。通过仔细选择条件，适体传感器在适合人类测试的水平（> 300 ng ml（-1））起作用，并且表现出类似于这种类型的基于抗体的传感器的灵敏度。适体的热稳定性显著优于相应的IgE抗体受体。此外，使用这种适体受体的传感器是稳健的（CV < 5%），并且每天变化很小（R.S.D. 8%）。回收率为100.5%或更好的两种类型的CNONA。最终传感器是一次性条带的方面使其成为将实验室环境的测定分散到即时护理设备的合适候选者。(c)2006 Elsevier B. V.保留所有权利。
The current work describes the systematic development of a disposable electrochemical sensor based on an aptamer receptor (aptasensor). The optimal conditions for which aptamers perform have been thoroughly investigated by Enzyme Linked OligoNucleic Assay (ELONA) and manipulated to achieve sufficient sensitivity in the final sensor set-up. Parameters investigated included buffer type, reagent concentration profiling, pH, ionic effect and thermo-stability. Method transfer then allowed for the preparation of the aptasensor utilising differential pulse voltammetry as the transduction mode. The principle system chosen was for the detection of allergy (IgE), which is present in humans due to exposure to a multitude of allergens in food and/or the environment. The system incorporates a competitive format for IgE detection using a biotinylated form of the aptamer. Enzymatic role is completed by the subsequent use of an extravidin-alkaline phosphatase label for either colorimetric, or electrochemical detection. Through the careful choice of conditions the aptasensor functions at levels suitable for human testing (> 300 ng ml(-1)) and exhibits sensitivity similar to antibody-based sensors of this type. The thermo-stability of the aptamer is significantly better than a corresponding IgE antibody receptor. In addition the sensor using this aptamer receptor is robust (CV < 5%) and varies little from day-to-day (R.S.D. 8%). Recoveries of 100 5% or better were obtained for two types of ELONA. The facet that the final sensor is a disposable strip makes it a suitable candidate for decentralisation of assaying away for the laboratory environment to a point-of-care device. (c) 2006 Elsevier B.V. All rights reserved.