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基于壳聚糖/SnO-SiC 空心球纳米链/金纳米材料的用于丙烯酰胺的超灵敏免疫传感器作为信号放大。

Ultrasensitive immunosensor for acrylamide based on chitosan/SnO-SiC hollow sphere nanochains/gold nanomaterial as signal amplification.

机构信息

Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University, Guangzhou, 510642, China; Foshan Polytechnic, Foshan, 528137, China.

Guangzhou Institute for Food Control, Guangzhou, 10410, China.

出版信息

Anal Chim Acta. 2019 Feb 21;1049:188-195. doi: 10.1016/j.aca.2018.10.041. Epub 2018 Oct 21.

DOI:10.1016/j.aca.2018.10.041
PMID:30612650
Abstract

An electrochemical immunosensor for ultrasensitive detection of acrylamide (AA) in water and food samples was developed. SnO-SiC hollow sphere nanochains with high surface area and gold nanoparticles with good electroconductivity were fabricated onto the surface of a glassy carbon electrode pre-coated with chitosan. The coating antigen (AA-4-mercaptophenylacetic acid-ovalbumin conjugate, AA-4-MPA-OVA) was immobilized on the electrode. Polyclonal antibody specific for AA-4-MPA was conjugated to gold nanorod (AuNR) as primary antibody (AuNR-Ab). Horseradish peroxidase labelled anti-rabbit antibody produced in goat was conjugated to AuNR as secondary antibody (HRP-AuNR-Ab). For detection, the analyte (AA-4-MPA) in sample competed with coating antigen for binding with AuNR-Ab. After washing, HRP-AuNR-Ab was added to capture the AuNR-Ab, and the electrical signal was obtained by addition of hydroquinone and HO. After investigation of the binding ability on nanomaterials and optimization of competitive immunoassay conditions, the proposed immunosensor exhibited a sensitive response to AA with a detection limit of 45.9 ± 2.7 ng kg, and working range of 187 ± 12.3 ng kg to 104 ± 8.2 μg kg for drinking water samples. Recoveries of AA from spiked samples were ranged from 86.0% to 115.0%. The specificity, repeatability and stability of the immunosensor were also proved to be acceptable, indicating its potential application in AA monitoring.

摘要

电化学免疫传感器用于超灵敏检测水中和食品样品中的丙烯酰胺 (AA)。SnO-SiC 空心球纳米链具有高比表面积,金纳米粒子具有良好的导电性,被制备到预先涂有壳聚糖的玻碳电极表面。将包被抗原 (AA-4-巯基苯乙酸-卵清蛋白偶联物,AA-4-MPA-OVA) 固定在电极上。针对 AA-4-MPA 的多克隆抗体与金纳米棒 (AuNR) 偶联作为一级抗体 (AuNR-Ab)。辣根过氧化物酶标记的山羊抗兔抗体与 AuNR 偶联作为二级抗体 (HRP-AuNR-Ab)。用于检测,样品中的分析物 (AA-4-MPA) 与包被抗原竞争与 AuNR-Ab 结合。洗涤后,加入 HRP-AuNR-Ab 捕获 AuNR-Ab,并通过加入对苯二酚和 HO 获得电信号。在研究了纳米材料的结合能力并优化了竞争性免疫分析条件后,该免疫传感器对 AA 表现出灵敏的响应,检测限为 45.9±2.7ngkg,工作范围为 187±12.3ngkg 至 104±8.2μgkg,用于饮用水样品。从加标样品中回收的 AA 范围为 86.0%至 115.0%。还证明了该免疫传感器的特异性、重复性和稳定性是可以接受的,表明其在 AA 监测中的潜在应用。

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