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基于一次性无试剂电化学免疫传感器阵列的电场驱动蛋白质生物标志物多重检测策略

Electric field-driven strategy for multiplexed detection of protein biomarkers using a disposable reagentless electrochemical immunosensor array.

作者信息

Wu Jie, Yan Yuetian, Yan Feng, Ju Huangxian

机构信息

Key Laboratory of Analytical Chemistry for Life Science (Ministry of Education of China), Department of Chemistry, Nanjing University, Nanjing 210093, PR China.

出版信息

Anal Chem. 2008 Aug 1;80(15):6072-7. doi: 10.1021/ac800905k. Epub 2008 Jul 2.

Abstract

A fast, simple, sensitive, and low-cost method for electrochemical multianalyte immunoassay was developed by combining newly designed electric field-driven incubation with a screen-printed reagentless immunosensor array. The disposable array was prepared by immobilizing respectively horseradish peroxidase (HRP)-labeled antibodies modified gold nanoparticles in biopolymer/sol-gel modified electrodes to obtain direct electrochemical responses of HRP. Upon the formation of immunocomplexes, the responses decreased due to increasing spatial blocking and impedance. At a driving potential of 0.5 V, the incubation process could be accomplished within 2 min. Under optimal conditions, this method could simultaneously detect carbohydrate antigens 153, 125, and 199 and carcinoembryonic antigens ranging from 0.084 to 16, 0.11 to 13, and 0.16 to 15 U mL(-1) and 0.16 to 9.2 ng mL(-1) with a detection time of less than 5 min, and the detection limits corresponding to the signals of 3SD were 0.06, 0.03, and 0.10 U mL(-1) and 0.04 ng mL(-1), respectively. The disposable immunosensor array and simple detection system for fast measurement of panels of tumor markers show significant clinical value for application in cancer screening and provide great potential for convenient point-of-care testing and commercial application.

摘要

通过将新设计的电场驱动孵育与丝网印刷无试剂免疫传感器阵列相结合,开发了一种用于电化学多分析物免疫测定的快速、简单、灵敏且低成本的方法。通过将辣根过氧化物酶(HRP)标记的抗体修饰的金纳米颗粒分别固定在生物聚合物/溶胶-凝胶修饰电极中,制备了一次性阵列,以获得HRP的直接电化学响应。免疫复合物形成后,由于空间位阻增加和阻抗增大,响应降低。在0.5 V的驱动电位下,孵育过程可在2分钟内完成。在最佳条件下,该方法可同时检测糖类抗原153、125和199以及癌胚抗原,检测范围分别为0.084至16、0.11至13和0.16至15 U mL(-1)以及0.16至9.2 ng mL(-1),检测时间小于5分钟,对应于3SD信号的检测限分别为0.06、0.03和0.10 U mL(-1)以及0.04 ng mL(-1)。这种用于快速测量肿瘤标志物组的一次性免疫传感器阵列和简单检测系统在癌症筛查应用中具有显著的临床价值,并为便捷的即时检测和商业应用提供了巨大潜力。

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