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基于含硅纳米粒子形成的用于荧光和比色测定的α-葡萄糖苷酶触发反应。

α-Glucosidase-Triggered Reaction for Fluorometric and Colorimetric Assays Based on the Formation of Silicon-Containing Nanoparticles.

机构信息

CAS Key Laboratory of Chemistry of Northwestern Plant Resources, Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences (CAS), Lanzhou 730000, P.R. China.

University of Chinese Academy of Sciences, Beijing 100049, P.R. China.

出版信息

Anal Chem. 2021 Nov 23;93(46):15412-15419. doi: 10.1021/acs.analchem.1c03210. Epub 2021 Nov 11.

DOI:10.1021/acs.analchem.1c03210
PMID:34762397
Abstract

Designing analytical approaches for enzymatic activity monitoring with high sensitivity and selectivity is of critical value for the diagnosis of diseases and biomedical studies. In this study, we have created a facile one-step synthetic route to prepare orange-red color and yellow fluorescent silicon-containing nanoparticles (Si CNPs) by mixing 3(2-aminoethylamino) propyl (dimethoxymethylsilane) and hydroquinone (HQ) in an aqueous solution. Inspired by the HQ-regulated facile synthetic step and the generation of HQ from α-glucosidase (α-Glu)-catalyzed hydrolysis of 4-hydroxyphenyl-α-d-glucopyranosyl (4-HPαDG), we have designed a straightforward colorimetric and fluorometric α-Glu activity assay using a commercially available 4-HPαDG as the α-Glu substrate. Fluorescent and colorimetric assays for α-Glu activity measurement have been thereby established and exhibited detection limits as low as 0.0032 and 0.0046 U/mL, respectively. Under single excitation at 370 nm, the prepared Si CNPs emitted yellow fluorescence at 520 nm and exhibited an absorbance peak at 390 nm. In addition, the proposed approach reveals various advantages including easy operation, time-saving, and good anti-interference ability. Hence, it could improve the progress of fluorometric and colorimetric enzymatic activity assays with high sensitivity and simplicity. Moreover, the proposed approach was applied for α-Glu inhibitor screening, and its feasibility in real samples was measured by detecting the α-Glu activity in human serum samples.

摘要

设计具有高灵敏度和选择性的酶活性监测分析方法对于疾病诊断和生物医学研究至关重要。在本研究中,我们通过在水溶液中混合 3(2-氨乙基氨基)丙基(二甲氧基甲基硅烷)和对苯二酚(HQ),开发了一种简便的一步合成方法来制备橙红色和黄色荧光含硅纳米粒子(SiCNPs)。受 HQ 调控的简便合成步骤以及 HQ 由α-葡萄糖苷酶(α-Glu)催化 4-羟基苯基-α-d-吡喃葡萄糖基(4-HPαDG)水解生成的启发,我们设计了一种使用市售的 4-HPαDG 作为α-Glu 底物的简单比色法和荧光法α-Glu 活性测定法。因此,建立了用于测量α-Glu 活性的荧光和比色测定法,检测限分别低至 0.0032 和 0.0046 U/mL。在 370nm 单激发下,制备的 SiCNPs 在 520nm 处发射黄色荧光,并在 390nm 处显示出吸收峰。此外,该方法具有操作简单、省时、抗干扰能力强等优点。因此,它可以提高荧光法和比色法酶活性测定的灵敏度和简单性。此外,该方法还用于α-Glu 抑制剂的筛选,并通过检测人血清样品中的α-Glu 活性来衡量其在实际样品中的可行性。

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