A Nanozyme- and Ambient Light-Based Smartphone Platform for Simultaneous Detection of Dual Biomarkers from Exposure to Organophosphorus Pesticides
A Nanozyme- and Ambient Light-Based Smartphone Platform for Simultaneous Detection of Dual Biomarkers from Exposure to Organophosphorus Pesticides
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DOI：
10.1021/acs.analchem.8b00837
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发表时间：
2018-06-19
影响因子：
7.4
通讯作者：
Du, Dan
中科院分区：
文献类型：
--
作者：
Zhao, Yuting;Yang, Mingming;Du, Dan
首先提出了一种与基于智能手机的环境光传感器耦合的透明侧向流测试条，用于检测酶抑制和磷酸化。该平台的原理是基于同时测量酶的总量和酶活性来生物监测有机磷农药的暴露。以丁酰胆碱酯酶（BChE）为模型酶，乙基对氧磷为有机磷农药的分析物。BChE的总量进行了定量的灵敏的比色信号源自夹心免疫层析测定，利用PtPd纳米粒子作为比色探针，表现出优异的催化活性酚。在夹心免疫分析中，由于天然BChE的四聚体结构，仅一种针对BChE的抗体同时用作识别抗体和标记抗体。使用Ellman测定通过另一个比色信号估计BChE活性。通过基于智能手机的环境光传感器检测两个单独的测试条上的两个比色信号。所提出的传感器在3D打印基板中使用LED进行操作，该基板发出激发光并将其传输通过透明的侧向流测试条。随着测试条的测试线中的比色信号的增加，透射光的强度降低。基于智能手机的传感器显示出优异的线性响应，用于测定BChE和活性BChE的总量，范围分别为0.05至6.4 nM和0.1至6.4 nM。在普通的基于智能手机的设备中同时实现了高便携性和低检测限。这种低成本、便携式、易于操作和灵敏的免疫测定策略显示出在线检测OP暴露和监测其他疾病生物标志物的巨大潜力。
A transparent, lateral-flow test strip coupled with a smartphone-based ambient light sensor was first proposed for detecting enzymatic inhibition and phosphorylation. The principle of the platform is based on the simultaneous measurement of the total amount of the enzyme and enzyme activity to biomonitor exposure to organophosphorus (OP) pesticides. In this study, butyrylcholinesterase (BChE) was adopted as the model enzyme and ethyl paraoxon was chosen as an analyte representing OP pesticides. The total amount of BChE was quantified by a sensitive colorimetric signal originating from a sandwich immunochromatographic assay utilizing PtPd nanoparticles as a colorimetric probe, which exhibited excellent catalytic activity for phenols. In the sandwich immunoassay, only one antibody against BChE was simultaneously utilized as the recognition antibody and the labeling antibody due to the tetrameric structure of native BChE. The BChE activity was estimated by another colorimetric signal using the Ellman assay. Both colorimetric signals on two separated test strips were detected by the smartphone-based ambient light sensor. The proposed sensor operated with an LED in a 3D-printed substrate, which emitted excitation light and transmitted it through a transparent, lateral-flow test strip. With the increase in the colorimetric signal in the test line of the test strip, the intensity of the transmitted light decreased. The smartphone-based sensor showed excellent linear responses for assaying the total amount of BChE and active BChE ranging from 0.05 to 6.4 nM and from 0.1 to 6.4 nM, respectively. A high portability and low detection limit were simultaneously realized in the common smartphone-based device. This low-cost, portable, easy-operation, and sensitive immunoassay strategy shows great potential for online detection of OP exposure and monitoring other disease biomarkers.