Label-free colorimetric detection platform based on catalytic hairpin self-assembly and G-quadruplex/hemin DNAzyme for comprehensive biomarker profiling

化学 脱氧核酶 血红素 G-四倍体 适体 核酸内切酶 生物标志物 纳米技术 荧光染料 生物传感器 组合化学 色谱法 检出限 分子生物学 实时聚合酶链反应 生物化学 血红素 DNA 生物 基因 材料科学
作者
Changjiang Li,Yuqiang Hu,Tianzi Shi,Kejun Dong,Tongbo Wu
出处
期刊:Talanta [Elsevier BV]
卷期号:272: 125835-125835 被引量:4
标识
DOI:10.1016/j.talanta.2024.125835
摘要

The expression level of human apurinic/apyrimidinic endonuclease 1 (APE1) is closely associated with the onset of various diseases, establishing it as a crucial clinical biomarker and a target in anti-cancer efforts. This study accomplished colorimetric and visual detection of APE1 by harnessing its endonuclease activity through catalytic hairpin self-assembly (CHA) and G-quadruplex/hemin DNAzyme. Optimization of the freedom degrees of the G-rich sequence significantly improved the detection performance of the strategy by influencing DNAzyme formation. Additionally, we replaced the signal reporting system with a molecular beacon to develop a fluorescence detection strategy, which served as an extension of the signal amplification system for validation and signal readout. The fluorescent probe method achieved a detection limit of 3.37 × 10−4 U/mL, while the colorimetric method yielded a detection limit of 6.5 × 10−3 U/mL, with a linear range spanning from 0.01 to 0.25 U/mL. Subsequently, the colorimetric approach effectively assessed APE1 activity in biological samples and facilitated the screening of APE1 activity inhibitors. Furthermore, this CHA/G-quadruplex/hemin DNAzyme strategy was adapted for the colorimetric detection of adenosine, showcasing its broad applicability across various biomarkers. The developed colorimetric analytical strategy represents a pivotal biosensing platform for diagnosing and treating diseases.
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