Chen Lizhen, Wu Jie, Yan Feng, Ju Huangxian
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, PR China.
Department of Applied Physics, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong.
Biosens Bioelectron. 2021 Mar 1;175:112878. doi: 10.1016/j.bios.2020.112878. Epub 2020 Dec 3.
This work designed a facile sensing strategy for quantitation of glycans on cell surface using organic electrochemical transistors (OECTs). The sensing strategy was performed by covalently binding target cells on mercaptopropionic acid modified gate electrode for the recognition of horseradish peroxidase (HRP) labeled lectins to specific glycans on cell surface, which led to sensitive channel current signal responding to the captured HRP and thus the expression of cell surface glycans. The quantitation of glycans on cell surface was achieved by using glycan modified microspheres to simulate the cells, on which the amounts of glycans were detected with enzyme linked immunosorbent assay. The proposed sensing strategy had been used for the detection of mannose and galactose on HeLa cells with Concanavalin A and peanut agglutinin as the specific lectins, which showed 1.37 × 10 mannose and 1.13 × 10 galactose on each cell, respectively. By treating HeLa cells with exoglycosidases, the mannose and galactose expression levels showed the same changes as those detected with flow cytometric analysis, indicating the practical application of the OECT-based biosensor in cell surface glycan expression analysis.
这项工作设计了一种简便的传感策略，用于使用有机电化学晶体管（OECT）对细胞表面的聚糖进行定量分析。该传感策略是通过将靶细胞共价结合在巯基丙酸修饰的栅电极上，以识别辣根过氧化物酶（HRP）标记的凝集素与细胞表面特定聚糖的结合，从而导致敏感的通道电流信号对捕获的HRP作出响应，进而反映细胞表面聚糖的表达。通过使用聚糖修饰的微球模拟细胞，利用酶联免疫吸附测定法检测其上的聚糖量，实现了对细胞表面聚糖的定量分析。所提出的传感策略已用于以伴刀豆球蛋白A和花生凝集素作为特异性凝集素检测HeLa细胞上的甘露糖和半乳糖，结果显示每个细胞上分别有1.37×10个甘露糖和1.13×10个半乳糖。通过用外切糖苷酶处理HeLa细胞，甘露糖和半乳糖的表达水平显示出与流式细胞术分析检测到的变化相同，这表明基于OECT的生物传感器在细胞表面聚糖表达分析中的实际应用。
