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基于多巴胺接枝二氧化硅纳米球的生物缀合物解体引发空穴捕获的溶菌酶高通量光电流型光电化学免疫分析

High-Throughput Signal-On Photoelectrochemical Immunoassay of Lysozyme Based on Hole-Trapping Triggered by Disintegrating Bioconjugates of Dopamine-Grafted Silica Nanospheres.

机构信息

Key Laboratory of Pesticide & Chemical Biology of Ministry of Education, College of Chemistry , Central China Normal University , Wuhan 430079 , P. R. China.

出版信息

ACS Sens. 2018 Aug 24;3(8):1480-1488. doi: 10.1021/acssensors.8b00253. Epub 2018 Jul 23.

Abstract

A unique split-type photoelectrochemical (PEC) immunoassay has been constructed for detection of low-abundance biocompounds (lysozyme, Lyz, used in this case) via a new trigger strategy by disintegrating bioconjugates of dopamine-grafted silica nanospheres (DA@SiONSs) for signal amplification. The preferred electron donor assembly of DA@SiONSs is first used as a molecular printboard for positioning anti-Lyz secondary antibody (Ab) through an amide reaction. With specific immunoreactions in a high-binding microplate, a sandwich immunoassay, the DA@SiONSs-based bioconjugate is achieved. By initiating the disintegration of the bioconjugates via acid etching, numerous electron donors of DA are released, thus efficiently triggering hole-trapping with amplified signals obtained. The smart integration of ZnInS-based heterojunctions as photoactive material, a split-type detection mode, and a new trigger strategy by disintegrating the DA@SiONSs-based bioconjugate offer an attractive high-throughput signal-on PEC immunoassay for detection of Lyz. Such an unusual PEC sensor exhibits an outstanding linear response to the concentration in the range between 0.002 and 500 ng mL, and the detection limit is as low as 0.6 ppt ( S/ N = 3). The as-fabricated assay is cost-effective and sensitive. It has been successfully used for measuring Lyz in real samples, which demonstrates great promise for practical applications.

摘要

一种独特的分体式光电化学(PEC)免疫分析已被构建,用于通过一种新的触发策略检测低丰度生物化合物(在这种情况下使用溶菌酶,Lyz),该策略通过分解多巴胺接枝硅纳米球(DA@SiONSs)的生物缀合物进行信号放大。DA@SiONSs 的首选电子供体组装首先被用作分子印版,通过酰胺反应定位抗 Lyz 二级抗体(Ab)。在高结合微孔板中进行特定的免疫反应,实现了基于 DA@SiONSs 的夹心免疫分析。通过酸蚀刻启动生物缀合物的分解,释放出大量的 DA 电子供体,从而有效地触发空穴捕获并获得放大的信号。基于 ZnInS 的异质结作为光活性材料的智能集成、分体式检测模式以及通过分解基于 DA@SiONSs 的生物缀合物的新触发策略,为 Lyz 的检测提供了一种有吸引力的、高通量的信号开启型 PEC 免疫分析。这种不寻常的 PEC 传感器对 0.002 至 500 ng mL 范围内的浓度表现出出色的线性响应,检测限低至 0.6 ppt(S/N = 3)。所构建的分析方法具有成本效益和灵敏度。它已成功用于测量真实样品中的 Lyz,这为实际应用展示了巨大的前景。

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