Single probe-based catalytic quantum dot FRET nanosensor for human alkyladenine DNA glycosylase detection

Meng Liu; Nan Zhong; Lingfei Zhang; Qian Zhang; Xiaorui Tian; Fei Ma; Chun-Yang Zhang

doi:10.1016/j.talanta.2023.125089

Single probe-based catalytic quantum dot FRET nanosensor for human alkyladenine DNA glycosylase detection

Talanta. 2024 Jan 1;266(Pt 2):125089. doi: 10.1016/j.talanta.2023.125089. Epub 2023 Aug 18.

Authors

Meng Liu¹, Nan Zhong¹, Lingfei Zhang², Qian Zhang¹, Xiaorui Tian¹, Fei Ma³, Chun-Yang Zhang⁴

Affiliations

¹ College of Chemistry, Chemical Engineering and Materials Science, Shandong Normal University, Jinan, 250014, China.
² Center for Disease Control and Prevention of Weihai City, Weihai, 264200, China.
³ School of Chemistry and Chemical Engineering, Southeast University, Nanjing, 211189, China. Electronic address: fei@seu.edu.cn.
⁴ College of Chemistry, Chemical Engineering and Materials Science, Shandong Normal University, Jinan, 250014, China; School of Chemistry and Chemical Engineering, Southeast University, Nanjing, 211189, China. Electronic address: zhangcy@seu.edu.cn.

PMID: 37604071
DOI: 10.1016/j.talanta.2023.125089

Abstract

Human alkyladenine DNA glycosylase (hAAG) is essential for repairing alkylated and deaminated bases, and it has become a prospective diagnosis biomarker and a therapeutic target for disease treatment. However, most of hAAG assays suffer from complicated reaction scheme, poor specificity, long assay time, and limited sensitivity. Herein, we report a novel single probe-based catalytic quantum dot (QD) Förster resonance energy transfer (FRET) nanosensor for simple and sensitive detection of hAAG activity. In this assay, hAAG induces the generation of 3' OH terminus via the excision of I base and the cleavage of AP site by APE1, subsequently initiating strand displacement reaction to produce numerous ssDNA signal probes. These probes can self-assemble on the QD surface to induce efficient FRET between QD and Cy5. This assay is very simple with the involvement of only a single probe for the achievement of both specific sensing and efficient signal amplification. Moreover, each signal probe contains multiple Cy5 moieties, and multiple signal probes can assemble on a single QD to greatly enhance the FRET efficiency. This nanosensor exhibits a detection limit of 3.60 × 10^-10 U/μL and it is suitable for measuring enzymatic kinetics, screening inhibitor, and quantifying cellular hAAG activity with single-cell sensitivity.

Keywords: Fluorescence resonance energy transfer; Human alkyladenine DNA glycosylase; Quantum dot; Single-molecule detection.

MeSH terms

Fluorescence Resonance Energy Transfer*
Humans
Prospective Studies
Quantum Dots*

Substances

3-methyladenine-DNA glycosylase
cyanine dye 5