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基于原位形成硅纳米粒子的荧光测定法测定β-葡萄糖醛酸酶。

Fluorometric assay based on the in situ formation of silicon nanoparticles for the determination of β-glucuronidase.

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, People's Republic of China.

University of Chinese Academy of Sciences, Beijing, 100049, People's Republic of China.

出版信息

Mikrochim Acta. 2022 Nov 2;189(11):436. doi: 10.1007/s00604-022-05528-7.

Abstract

As a prodrug-converting enzyme, β-glucuronidase (β-GCase) is a lysosomal enzyme participating in the release of glucose from glucopyranosyl glycoside. In this work, for the first time, we have developed an analytical method exhibiting fluorometric signals for straightforward determination of β-GCase using silicon nanoparticles (Si NPs). Via hydrothermal treatment, in the water bath of 70 °C for 50 min, dopamine (DA) reacts with (3-[2-(2-aminoethylamino) ethylamino] propyltrimethoxysilane) (AEEA) to produce green fluorescent Si NPs. Enlightened by such easy reaction and β-GCase-triggered specific hydrolysis of dopamine-4-β-D-glucuronide (DA-GCU) into DA, we have designed an analytical method for β-GCase sensing through the production of Si NPs. Therefore, through the designed sensing platform, β-GCase activity was monitored, and the limit of detection (LOD) for this study was 0.02 U/L. Furthermore, the feasibility of the method was assessed by measuring β-GCase activity in human serum where recoveries and RSD were in the ranges 99-104% and 1.37-3.44, respectively.

摘要

作为一种前体药物转化酶,β-葡糖苷酸酶(β-GCase)是一种溶酶体酶,参与葡萄糖从吡喃葡萄糖苷糖苷中的释放。在这项工作中,我们首次开发了一种分析方法,该方法使用硅纳米粒子(Si NPs)显示出荧光信号,可直接测定β-GCase。通过水热处理,在 70°C 的水浴中反应 50 分钟,多巴胺(DA)与(3-[2-(2-氨乙基氨基)乙基氨基]丙基三甲氧基硅烷)(AEEA)反应生成绿色荧光 Si NPs。受这种简单反应和β-GCase 触发的多巴胺-4-β-D-葡糖苷酸(DA-GCU)特异性水解的启发,我们设计了一种通过 Si NPs 生成来检测β-GCase 的分析方法。因此,通过设计的传感平台,监测了β-GCase 的活性,本研究的检测限(LOD)为 0.02 U/L。此外,通过测量人血清中的β-GCase 活性评估了该方法的可行性,回收率和 RSD 的范围分别为 99-104%和 1.37-3.44。

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