Plasmonic Gold Nanostar-Based Probes with Distance-Dependent Plasmon-Enhanced Fluorescence for Ultrasensitive DNA Methyltransferase Assay,Analytical Chemistry

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Plasmonic Gold Nanostar-Based Probes with Distance-Dependent Plasmon-Enhanced Fluorescence for Ultrasensitive DNA Methyltransferase Assay
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-03-08 , DOI: 10.1021/acs.analchem.3c04122
Tingting Zhao ₁ , Xiaozhe Pang ₁ , Congkai Wang ₁ , Lei Wang ₁ , Yifan Yang _{1,

2} , Junqi Wang ₁ , Jiangfei Jia ₁ , Xinxue Liu ₁ , Shenghao Xu ₁ , Xiliang Luo ₁

Affiliation

The ultrasensitive DNA methyltransferase (Dam MTase) assay is of high significance for biomedical research and clinical diagnosis because of its profound effect on gene regulation. However, detection sensitivity is still limited by shortcomings, including photobleaching and weak signal intensities of conventional fluorophores at low concentrations. Plasmonic nanostructures with ultrastrong electromagnetic fields and fluorescence enhancement capability that can overcome these intrinsic defects hold great potential for ultrasensitive bioanalysis. Herein, a silica-coated gold nanostars (Au NSTs@SiO₂)-based plasmon-enhanced fluorescence (PEF) probe with 20 “hot spots” was developed for ultrasensitive detection of Dam MTase. Here, the Dam Mtase assay was achieved by detecting the byproduct PPi of the rolling circle amplification reaction. It is worth noting that, benefiting from the excellent fluorescence enhancement capability of Au NSTs originating from their 20 “hot spots”, the detection limit of Dam Mtase was reduced by nearly 105 times. Moreover, the proposed Au NST-based PEF probe enabled versatile evaluation of Dam MTase inhibitors as well as endogenous Dam MTase detection in GW5100 and JM110 Escherichia coli cell lysates, demonstrating its potential in biomedical analysis.

中文翻译：

基于等离激元金纳米星的探针，具有距离依赖性等离激元增强荧光，用于超灵敏 DNA 甲基转移酶测定

超灵敏DNA甲基转移酶（Dam MTase）检测由于其对基因调控的深远影响，对于生物医学研究和临床诊断具有重要意义。然而，检测灵敏度仍然受到缺点的限制，包括光漂白和低浓度下常规荧光团的信号强度弱。具有超强电磁场和荧光增强能力的等离子体纳米结构可以克服这些固有缺陷，在超灵敏生物分析方面具有巨大的潜力。在此，开发了一种具有 20 个“热点”的基于二氧化硅涂层金纳米星 (Au NSTs@SiO ₂ ) 的等离子体增强荧光 (PEF) 探针，用于超灵敏检测 Dam MTase。在这里，Dam Mtase 测定是通过检测滚环扩增反应的副产物 PPi 来实现的。值得注意的是，得益于Au NSTs源自其20个“热点”的优异荧光增强能力，Dam Mtase的检测限降低了近105倍。此外，所提出的基于 Au NST 的 PEF 探针能够对 Dam MTase 抑制剂进行多功能评估，并能够检测 GW5100 和 JM110大肠杆菌细胞裂解物中的内源性 Dam MTase，展示了其在生物医学分析中的潜力。

更新日期：2024-03-08

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