Determination of membrane PD-L1 by SECM technique based on aptamer identification

IF 4.1 3区化学 Q1 CHEMISTRY, ANALYTICAL Journal of Electroanalytical Chemistry Pub Date : 2025-02-07 DOI:10.1016/j.jelechem.2025.118999

Yuying Du, Ziqi Wang, Jiening Wu, Liping Lu

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Abstract

Membrane proteins play crucial roles in cellular activities and are the major actors of bio-membrane functions. Programmed death ligand receptor 1 (PD-L1) is a type of transmembrane protein that is overexpressed on certain tumor cells, leading to the immune escape of cancer cells. Here, a detection method was developed using scanning electrochemical microscopy (SECM) through aptamer-specific recognition and enzyme-catalyzed reaction, which converts the PD-L1 expression into an electrical signal. The aptamer (MJ5C) modified alkaline phosphatase (ALP) can specifically capture PD-L1, and ALP catalyzes the reduction of 4-aminophenyl phosphate (PAPP) to p-aminophenol (PAP), the current response of PAP at SECM tip is positively correlated with the expression of PD-L1 on NCI-H1975 cell. The results showed that this method could real-time detect the expression of PD-L1 on a single cell stimulated by drugs and dibenzothiophene (DBT). Overall, this method provides a new feasible method for the real-time nondestructive detection of single-cell membrane protein expression and a new avenue for studying the effect of pollutants on membrane protein.

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基于适体鉴定的膜PD-L1的SECM技术测定

膜蛋白在细胞活动中起着至关重要的作用，是生物膜功能的主要参与者。程序性死亡配体受体1 （Programmed death ligand receptor 1, PD-L1）是一种跨膜蛋白，在某些肿瘤细胞上过表达，导致癌细胞免疫逃逸。本研究利用扫描电化学显微镜（SECM）通过适配体特异性识别和酶催化反应，将PD-L1表达转化为电信号，建立了一种检测方法。适配体（MJ5C）修饰的碱性磷酸酶（ALP）可以特异性捕获PD-L1， ALP催化4-氨基苯基磷酸（PAPP）还原为对氨基酚（PAP）， PAP在SECM顶端的当前反应与NCI-H1975细胞上PD-L1的表达呈正相关。结果表明，该方法可以实时检测药物和二苯并噻吩（DBT）刺激下单细胞上PD-L1的表达。总之，该方法为实时无损检测单细胞膜蛋白表达提供了一种新的可行方法，也为研究污染物对膜蛋白的影响提供了新的途径。

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来源期刊

Journal of Electroanalytical Chemistry 化学-电化学

CiteScore

7.80

自引率

6.70%

发文量

912

审稿时长

2.4 months

期刊介绍： The Journal of Electroanalytical Chemistry is the foremost international journal devoted to the interdisciplinary subject of electrochemistry in all its aspects, theoretical as well as applied. Electrochemistry is a wide ranging area that is in a state of continuous evolution. Rather than compiling a long list of topics covered by the Journal, the editors would like to draw particular attention to the key issues of novelty, topicality and quality. Papers should present new and interesting electrochemical science in a way that is accessible to the reader. The presentation and discussion should be at a level that is consistent with the international status of the Journal. Reports describing the application of well-established techniques to problems that are essentially technical will not be accepted. Similarly, papers that report observations but fail to provide adequate interpretation will be rejected by the Editors. Papers dealing with technical electrochemistry should be submitted to other specialist journals unless the authors can show that their work provides substantially new insights into electrochemical processes.