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基于适配体共价连接到多壁碳纳米管/离子液体/壳聚糖纳米复合材料修饰电极的无标记电化学 IgE 适体传感器。

Label-free electrochemical IgE aptasensor based on covalent attachment of aptamer onto multiwalled carbon nanotubes/ionic liquid/chitosan nanocomposite modified electrode.

机构信息

Department of Chemistry, University of Kurdistan, P. O. Box 416, Sanandaj, Iran.

出版信息

Biosens Bioelectron. 2013 May 15;43:218-25. doi: 10.1016/j.bios.2012.12.006. Epub 2012 Dec 20.

Abstract

A simple, sensitive and label-free aptamer-based biosensor for the detection of human immunoglobulin E (IgE) is developed using the electrochemical transduction method. A special immobilization interface consisting of multiwalled carbon nanotubes/ionic liquid/chitosan nanocomposite (MWCNTs/IL/Chit) is utilized to improve the conductivity and performance characteristics of the biosensor as well as to increase the loading amount of aptamer DNA sequence. A 5'-amino-terminated aptamer is covalently attached onto MWCNTs/IL/Chit modified glassy carbon (GC) electrode via a linker of glutaraldehyde (GA). Methylene blue (MB) is used as an electrochemical indicator which is intercalated into the aptamer through the specific interaction with its guanine bases. In the absence of IgE, MB bound to the aptamer produces a strong differential pulse voltammetric (DPV) signal. But when IgE exists, the intercalated MB releases from the aptamer, resulting an obviously decreased DPV signal. This phenomenon can be applied for human IgE detection. The peak current of MB linearly decreases with the concentration of IgE over a range of 0.5-30 nM with a detection limit of 37 pM. By using Bovine serum albumin (BSA) and lysozyme, the excellent specificity of this sensing system for the detection of IgE is also demonstrated. Finally, the proposed aptasensor is successfully used to IgE analysis in human serum sample. The obtained result is well agreed with the value obtained by the standard ELISA method. The herein described approach is expected to promote the exploitation of aptamer-based biosensors for protein assays in biochemical and biomedical studies.

摘要

基于适配体的电化学生物传感器可用于检测人免疫球蛋白 E(IgE),本研究开发了一种简单、灵敏、无需标记的检测方法。该方法利用电化学转导技术,采用多壁碳纳米管/离子液体/壳聚糖纳米复合材料(MWCNTs/IL/Chit)特殊固定化界面,以提高传感器的导电性和性能特点,并增加适配体 DNA 序列的负载量。5'-氨基末端的适配体通过戊二醛(GA)连接子共价连接到 MWCNTs/IL/Chit 修饰的玻碳电极上。亚甲基蓝(MB)用作电化学指示剂,通过与碱基的特异性相互作用插入适配体中。在不存在 IgE 的情况下,与适配体结合的 MB 产生强差分脉冲伏安法(DPV)信号。但是,当存在 IgE 时,插入的 MB 从适配体中释放出来,导致 DPV 信号明显降低。这种现象可用于人 IgE 的检测。MB 的峰电流随 IgE 浓度在 0.5-30 nM 范围内呈线性下降,检测限为 37 pM。通过使用牛血清白蛋白(BSA)和溶菌酶,还证明了该传感系统对 IgE 检测具有优异的特异性。最后,该适配体传感器成功用于人血清样品中 IgE 的分析。得到的结果与标准 ELISA 方法获得的值非常吻合。该方法有望促进适配体生物传感器在生化和生物医学研究中用于蛋白质分析的开发。

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