TCM and Ethnomedicine Innovation & Development International Laboratory, School of Pharmacy, Hunan University of Chinese Medicine, 410208, Changsha, China.
The First Affiliated Hospital of Hunan University of Traditional Chinese Medicine, 410021, Changsha, China.
Talanta. 2024 Dec 1;280:126758. doi: 10.1016/j.talanta.2024.126758. Epub 2024 Aug 24.
DNA walkers have emerged as a powerful tool in various biosensors, enabling the detection of low-abundance analytes with their precise programmability and efficient signal amplification capacity. However, many existing approaches are hampered by limited reaction kinetics. Herein, we designed a stochastic bipedal dual-DNA walkers (SBDW) that can traverse at high speed on AuNP-based three-dimensional (3D) tracks powered by Exo III. The SBDW exhibited superior reaction kinetics and are up to least 2.25 times faster than traditional DNA walkers, reaching a plateau within 40 min. This advancement allows for rapid and highly sensitive fluorescence detection of a significant base excision repair enzyme of APE1 with a detection limit of 0.001 U/mL. In comparison to traditional DNA walkers, this platform enables highly sensitive and specific APE1 assays in cell lysate and facilitates rapid and accurate screening of APE1 inhibitors. Given its rapid, sensitive, specific, and reliable analysis features, the strategy shows great promise in drug discovery and clinical diagnosis.
DNA walker 已成为各种生物传感器中的强大工具，其精确的可编程性和高效的信号放大能力使其能够检测低丰度分析物。然而，许多现有方法受到有限的反应动力学的限制。在此，我们设计了一种随机双足双 DNA walker (SBDW)，它可以在 Exo III 驱动的基于 AuNP 的三维 (3D) 轨道上高速穿越。SBDW 表现出优越的反应动力学，比传统的 DNA walker 快至少 2.25 倍，在 40 分钟内达到平台期。这一进展使得可以快速且高度灵敏地检测 APE1 的一个重要碱基切除修复酶，检测限为 0.001 U/mL。与传统的 DNA walker 相比，该平台能够在细胞裂解物中进行高度敏感和特异的 APE1 分析，并促进 APE1 抑制剂的快速准确筛选。鉴于其快速、灵敏、特异和可靠的分析特点，该策略在药物发现和临床诊断中具有广阔的应用前景。
