通过杂交链式反应提高球形核酸酶的催化活性及其在黄曲霉毒素 B1 灵敏度分析中的应用

Wenjun Wang, Xuesong Li, Kun Zeng, Yanyan Lu, Boyuan Jia, Jianxia Lv, Chenghao Wu, Xinyu Wang, Xinshuo Zhang, Zhen Zhang
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引用次数: 0

摘要

传统的球形核酸酶(SNAzymes)以金纳米粒子(AuNPs)为核心,以DNA为外壳,因其优异的物理化学特性而广泛应用于生物分析领域。尽管如此,有限的 AuNPs 表面上拥挤的催化单元(如 G-四联体,G4)不可避免地会影响其催化活性。因此,我们采用杂交链反应(HCR)来扩大 G4 酶的数量和空间,从而提高其催化能力。通过系统研究,我们发现当不完整的 G4 序列连接在具有分裂模式(3:1 和 2:2)的发夹的粘性末端时,由于增加了立体阻碍,会显著降低 HCR 杂交能力。相反,在发夹的非粘性末端直接修饰完整的 G4 序列后,由于避免了立体阻碍,HCR 杂交能力明显增强。因此,我们将利用 HCR 改进的 SNAzymes 应用于食品样品中 AFB1 的检测,作为概念验证,结果表明其性能优异(检测限为 0.08 ng/mL)。重要的是,我们的策略为利用 G4 作为信号分子提高 SNAzymes 的催化活性提供了新的思路。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Improved Catalytic Activity of Spherical Nucleic Acid Enzymes by Hybridization Chain Reaction and Its Application for Sensitive Analysis of Aflatoxin B1
Conventional spherical nucleic acid enzymes (SNAzymes), made with gold nanoparticle (AuNPs) cores and DNA shells, are widely applied in bioanalysis owing to their excellent physicochemical properties. Albeit important, the crowded catalytic units (such as G-quadruplex, G4) on the limited AuNPs surface inevitably influence their catalytic activities. Herin, a hybridization chain reaction (HCR) is employed as a means to expand the quantity and spaces of G4 enzymes for their catalytic ability enhancement. Through systematic investigations, we found that when an incomplete G4 sequence was linked at the sticky ends of the hairpins with split modes (3:1 and 2:2), this would significantly decrease the HCR hybridization capability due to increased steric hindrance. In contrast, the HCR hybridization capability was remarkably enhanced after the complete G4 sequence was directly modified at the non-sticky end of the hairpins, ascribed to the steric hindrance avoided. Accordingly, the improved SNAzymes using HCR were applied for the determination of AFB1 in food samples as a proof-of-concept, which exhibited outstanding performance (detection limit, 0.08 ng/mL). Importantly, our strategy provided a new insight for the catalytic activity improvement in SNAzymes using G4 as a signaling molecule.
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