Investigating lysozyme amyloid fibrillization by electrochemical impedance spectroscopy for application in lysozyme sensor

化学 溶菌酶 介电谱 纤维 生物物理学 电化学 电极 生物化学 物理化学 生物
作者
Ibrar Alam,Benchaporn Lertanantawong,Worapot Prongmanee,Tossaporn Lertvanithphol,Mati Horprathum,Thana Sutthibutpong,Piyapong Asanithi
出处
期刊:Journal of Electroanalytical Chemistry [Elsevier BV]
卷期号:901: 115799-115799 被引量:8
标识
DOI:10.1016/j.jelechem.2021.115799
摘要

Amyloid fibrils have garnered increasing attention as viable building blocks for designing and synthesizing of biomedical material. The structure of amyloid fibril of most of the proteins are well known, but the electrical property of their conformational changes during synthesis is not fully understood. Herein, we investigated this relationship by using non-Faradaic electrochemical impedance spectroscopy (EIS). Lysozyme (Lyz) from hen egg white was used as a model protein and incubated under heat (60 °C) at pH 2 which led to the amyloid fibrillization. Morphological changes during fibrillization were confirmed by AFM. Raman spectroscopy was used to distinguish amyloid fibrils from their native and amorphous structures. For EIS measurement, graphene oxide coated screen-printed electrode (GO/SPE) was immobilized with Lyz aggregates obtained from different incubation times. The charge transfer resistance (Rct) of incubated Lyz was measured to determine the fibrillization pathway of amyloid fibril i.e., unfolding and nucleation, protofibril (PF, a short fibril) and mature fibril. Next, PF-modified GO/SPE (PF/GO/SPE) was demonstrated to determine Lyz molecules dissolved in PBS and in wine using EIS. The dependence of Rct of PF/GO/SPE on Lyz concentration was reported. Linear range and limit of detection (LOD) of detecting Lyz molecules in PBS and in wine were the same. The linear range and LOD were 0.005–0.08 μM and 0.001 μM, respectively. PF/GO/SPE sensor was found to have good selectivity to recognize Lyz. These findings show that PF/GO/SPE sensor provides an attractive component for future electrochemical sensing devices.
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