<br>目标驱动的环形 DNA Walker 与电化学发光硅量子点相结合用于 APE1 生物分析,Analytical Chemistry

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目标驱动的环形 DNA Walker 与电化学发光硅量子点相结合用于 APE1 生物分析
Analytical Chemistry ( IF 6.7 ) Pub Date : 2023-12-06 , DOI: 10.1021/acs.analchem.3c04747
Ying-Huan Li ₁ , Mei-Ling Zhao ₁ , Pu Zhang ₁ , Xia Yang ₁ , Ruo Yuan ₁ , Ying Zhuo ₁

Affiliation

具有大量纳米结构的功能性 DNA 行走器已被广泛研究；然而，当它们暴露于临床生物样本中的多种核酸酶时，其稳定性仍然面临挑战，导致实际评估的不可靠性。这项工作提出了一种具有增强稳定性的靶标驱动的环形 DNA walker，通过制备硅量子点 (SiQD) 作为电化学发光 (ECL) 发射器，能够对不同浓度的脱嘌呤/脱嘧啶核酸内切酶 1 (APE1) 做出灵敏可靠的响应。具体来说，通过透析去除 SiQD 周围的凝胶状层来纯化 SiQD 的微环境，SiQD 显示出显着强且稳定的 ECL 信号。与未经透析的 SiQD 相比，连续扫描下 ECL 信号的相对标准偏差 (RSD) 提高了 16.59 倍，表明 ECL 稳定性显着提高。随后，在 APE1 存在的情况下，设计的环形 DNA walker 被激活，沿着连续交叉 DNA 轨道内的众多淬灭探针移动，这些探针固定在 SiQD 修饰的电极上，提供 ECL 读出信号。该ECL生物传感器的线性范围为1.0 × 10 ^–13 U·μL ^–1至1.0 × 10 ^–7 U·μL ^–1 ，检测限(LOD)低至1.766 × 10 ^–14 U·μL ^–1 。该工作为临床样本检测提供了一种具有核酸酶抗性的DNA walker的新颖结构，并设计了合成具有良好ECL性能的SiQD的新策略，极大地扩展了SiQD的ECL应用。

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Target-Driven Annular DNA Walker Coupled with Electrochemiluminescent Silicon Quantum Dots for APE1 Bioanalysis

Functional DNA walkers with substantial nanostructures have been extensively investigated; however, their stability still faces challenges when exposed to diverse nuclease in clinical biological samples, resulting in the unreliability of actual assessment. This work proposed a target-driven annular DNA walker with enhanced stability enabling the sensitive and reliable response to different concentrations of apurinic/apyrimidinic endonuclease 1 (APE1), by preparing silicon quantum dots (SiQDs) as electrochemiluminescence (ECL) emitters. Specifically, the SiQDs showed significant strong and stable ECL signals by purifying the microenvironment of SiQDs through the dialysis removal of the gel-like layers surrounding the SiQDs. The relative standard deviation (RSD) of their ECL signal had been improved 16.59 times under consecutive scanning compared to that of SiQDs without dialysis, demonstrating a significant improvement in ECL stability. Subsequently, in the presence of APE1, the designed annular DNA walker was activated to move along the numerous quenching probes within the continuous cross-based DNA orbits, which were immobilized to the SiQD-modified electrode, providing ECL readout signals. The linear range of this ECL biosensor was 1.0 × 10^–13 U·μL^–1 to 1.0 × 10^–7 U·μL^–1, and the limit of detection (LOD) was as low as 1.766 × 10^–14 U·μL^–1. This work provides a novel structure of a DNA walker with nuclease resistance for clinical sample detection and designs a new strategy for synthesizing SiQDs with favorable ECL performance, tremendously expanding the ECL application of SiQDs.

更新日期：2023-12-06

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