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基于肽的 QCM 生物传感器用于胰蛋白酶的无标记分析性能。

Label-free analytical performances of a peptide-based QCM biosensor for trypsin.

机构信息

College of Environmental Science and Engineering, Anhui Normal University, Wuhu 241000, PR China.

出版信息

Analyst. 2020 May 7;145(9):3329-3338. doi: 10.1039/d0an00308e. Epub 2020 Mar 24.

Abstract

A label-free biosensor was fabricated for the detection of trypsin by using a peptide-functionalized quartz crystal microbalance gold electrode. The synthetized peptide chains were immobilized tightly on the QCM electrode via a self-assembly method, which formed a thin and approximate rigid layer of peptides. The detection signal was achieved by calculating the mass changes on the QCM electrode because the peptide chains could be specifically cleaved in the carboxyl terminuses of arginine and lysine by trypsin. When gold nanoparticles were coupled to the peptide chains, the sensing signal would be amplified 10.9 times. Furthermore, the sensor interface shows a lower resonance resistance change when the peptide chain is immobilized horizontally. Independent detections in parallel on different electrodes have a wide linear range. Under the optimum conditions, the signal-amplified biosensor allowed the measurement of trypsin over the range of 0-750 ng mL with a detection limit of 8.6 ng mL. Moreover, for screening the inhibitor of trypsin, the IC values were obtained to be 1.85 μg mL for benzamidine hydrochloride and 20.5 ng mL for the inhibitor from soybean.

摘要

一种无标记的生物传感器通过使用肽功能化的石英晶体微天平金电极来检测胰蛋白酶。合成的肽链通过自组装方法紧密固定在 QCM 电极上,形成了一层薄且近似刚性的肽层。检测信号是通过计算 QCM 电极上的质量变化来实现的,因为肽链可以通过胰蛋白酶在精氨酸和赖氨酸的羧基末端特异性切割。当金纳米粒子与肽链偶联时,传感信号会放大 10.9 倍。此外,当肽链水平固定时,传感器接口显示出较低的共振电阻变化。在不同电极上平行进行独立检测具有较宽的线性范围。在最佳条件下,信号放大生物传感器允许在 0-750ng/mL 的范围内测量胰蛋白酶,检测限为 8.6ng/mL。此外,为了筛选胰蛋白酶抑制剂,获得了苯甲脒盐酸盐的 IC 值为 1.85μg/mL,大豆抑制剂的 IC 值为 20.5ng/mL。

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