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一种基于微流控电化学适体传感器,通过 IDAs 适体,对含有整合素 α6β4 的 A549 细胞作为整合素 α6β4 细胞模型进行高灵敏度和选择性检测。

A microfluidic electrochemical aptasensor for highly sensitive and selective detection of A549 cells as integrin α6β4-containing cell model via IDA aptamers.

机构信息

Department of Medicinal Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran; Laboratory of Microfluidics and Medical Microsystems, BuAli Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran.

Laboratory of Microfluidics and Medical Microsystems, BuAli Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Mechanical Engineering, Ferdowsi University of Mashhad, Mashhad, Iran.

出版信息

Talanta. 2023 Jan 15;252:123781. doi: 10.1016/j.talanta.2022.123781. Epub 2022 Aug 6.

Abstract

There is a growing interest in developing microfluidic biosensors for the accurate and reproducible analysis of various biomarkers obtained from liquid biopsy. This paper reports a novel microfluidic electrochemical aptasensor for determination of A549 cells as integrin α6β4-containing cell model and circulating tumor cell (CTC) model of NSCLC, based on target-induced structure switching mode. The conformational change of IDA aptamer structure with an affinity towards A549 cells, in the absence and presence of A549 cells allowed selective and sensitive detection of A549 cells. The microfluidic biosensor consisted of a microchip integrated with a screen-printed gold electrode functionalized with SH-IDA aptamers via covalent chemistry. Target solution containing A549 cells in Phosphate-buffered saline could be introduced to the microchip for analysis. Upon exposing to redox probe, a reduction in peak current occurred. Required flow sequencing for various steps of the detection protocol was performed using on-chip gas-actuated microvalves with programmable operation. The microfluidic biosensor provided a wide linear dynamic range of 50-5 × 10 cells/mL and allowed determination of A549 cells with a detection limit of 14 cells/mL. Furthermore, the microfluidic biosensor was efficiently used for detection of A549 cells in complex matrices such as human serum. Our novel microfluidic aptasensor presents an enabling analytical platform to perform various detections of low volume biomarkers with high accuracy and reproducibility.

摘要

人们越来越感兴趣的是开发微流控生物传感器,以准确和可重复地分析从液体活检中获得的各种生物标志物。本文报道了一种新颖的微流控电化学适体传感器,用于基于靶标诱导结构转换模式测定 A549 细胞作为整合素 α6β4 含细胞模型和非小细胞肺癌的循环肿瘤细胞(CTC)模型。在不存在和存在 A549 细胞的情况下,IDA 适体结构的构象变化对 A549 细胞具有亲和力,允许对 A549 细胞进行选择性和灵敏检测。微流控生物传感器由一个微芯片组成,该微芯片通过共价化学将 SH-IDA 适体集成到功能化的丝网印刷金电极上。含有 A549 细胞的磷酸盐缓冲盐水的靶溶液可被引入微芯片进行分析。在暴露于氧化还原探针时,峰电流发生减少。使用带有可编程操作的片上气动微阀对检测方案的各个步骤进行所需的流动排序。微流控生物传感器提供了 50-5×10^3 个细胞/mL 的宽线性动态范围,并允许以 14 个细胞/mL 的检测限测定 A549 细胞。此外,该微流控生物传感器有效地用于检测人血清等复杂基质中的 A549 细胞。我们的新型微流控适体传感器提供了一个启用的分析平台,可高精度和可重复性地执行各种低体积生物标志物的检测。

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