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一种基于免疫磁分离和液滴阵列的用于自动检测癌胚抗原的微流控免疫传感器。

A microfluidic immunosensor for automatic detection of carcinoembryonic antigen based on immunomagnetic separation and droplet arrays.

机构信息

School of Ophthalmology & Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou, 325027, China.

State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China.

出版信息

Analyst. 2023 May 2;148(9):1939-1947. doi: 10.1039/d2an01922a.

Abstract

Diagnosis of cancer by biomarkers plays an important role in human health and life. However, current laboratory techniques for detecting cancer biomarkers still require laborious and time-consuming operation by skilled operators and associated laboratory instruments. This work presents a colorimetric biosensor for the rapid and sensitive detection of carcinoembryonic antigen (CEA) based on an automated immunomagnetic separation platform and a droplet array microfluidic chip with the aid of an image analysis system. Immunomagnetic nanoparticles (MNPs) were used to capture CEA in the samples. CEA-detecting antibodies and horseradish peroxidase (HRP) were modified on polystyrene microspheres (PS), catalysing hydrogen peroxide and 3,3',5,5'-tetramethylbenzidine (TMB) as signal outputs. Color reaction data were analyzed to establish a CEA concentration standard curve. The movement of MNPs between droplets in the microfluidic chip is achieved using an automatically programmable magnetic control system. This colorimetric biosensor has been used for the simultaneous detection of six CEA samples ranging from 100 pg mL to 100 ng mL with a detection limit of 14.347 pg mL in 10 min, following the linear equation: = -4.773 ln() + 156.26 with a correlation of = 0.9924, and the entire workflow can be completed within 80 minutes. The microfluidic immunosensor designed in this paper has the advantages of low cost, automation, low sample consumption, high throughput, and promising applications in biochemistry.

摘要

基于自动化免疫磁分离平台和液滴阵列微流控芯片,结合图像分析系统,本工作提出了一种用于快速灵敏检测癌胚抗原(CEA)的比色生物传感器。免疫磁珠(MNPs)用于捕获样品中的 CEA。CEA 检测抗体和辣根过氧化物酶(HRP)被修饰在聚苯乙烯微球(PS)上,作为信号输出物催化过氧化氢和 3,3',5,5'-四甲基联苯胺(TMB)的反应。通过分析颜色反应数据建立 CEA 浓度标准曲线。使用自动可编程磁控制系统实现 MNPs 在微流控芯片液滴之间的运动。该比色生物传感器用于同时检测 6 个 CEA 样本,浓度范围为 100 pg mL 至 100 ng mL,检测限为 14.347 pg mL,在 10 min 内,线性方程为:= -4.773 ln() + 156.26,相关系数为 = 0.9924,整个工作流程可在 80 分钟内完成。本文设计的微流控免疫传感器具有成本低、自动化、低样品消耗、高通量等优点,在生物化学领域具有广阔的应用前景。

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