<br>通过底物竞争性和仿生级联反应与高活性 NADH 氧化酶模拟物进行创新的比色 NQO1 检测策略,Analytical Chemistry

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通过底物竞争性和仿生级联反应与高活性 NADH 氧化酶模拟物进行创新的比色 NQO1 检测策略
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-07-30 , DOI: 10.1021/acs.analchem.4c03003
Qi Fang _{1,

2} , Quanyi Liu _{1,

2} , Zhimin Song _{1,

2} , Yu Wang ₃ , Xiaojun Zhang ₁ , Jun Cao ₄ , Jian Sun ₃ , Chong-Bo Ma ₄ , Yan Du _{1,

2}

Affiliation

NAD(P)H：醌氧化还原酶-1 (NQO1) 在抗氧化中发挥关键作用，并在肿瘤中异常过度表达。开发一种快速、灵敏的 NQO1 监测方法将极大地促进临床实践中的癌症诊断。本研究引入了 NQO1 的变革性比色检测策略，利用 NQO1 和新型 NADH 氧化酶 (NOX) 模拟物、钴氮掺杂碳纳米酶 (CoNC) 之间的创新竞争底物机制。该方法巧妙地利用 NQO1 存在下 NADH 的差异消耗来调节 CoNC 催化中 H ₂ O ₂的生成，然后通过涉及普鲁士蓝 (PB) 纳米粒子的二级过氧化物酶模拟级联反应进行定量。这种双阶段反应框架不仅提高了 NQO1 检测的灵敏度，实现低至 0.67 μg mL ^–1的检测限，而且还能够通过酶活性谱区分癌细胞和非癌细胞。此外，CoNC 表现出卓越的催化效率，比活性达到 5.2 U mg ^–1 ，明显优于现有的 NOX 模拟物。除了单纯的检测之外，CoNC 还具有双重作用，既充当细胞色素 c 还原酶 (Cyt c) 的强大模拟物，又充当酶再生的基石，从而扩大了其生物应用范围。这项研究不仅标志着纳米酶的生物分析应用向前迈出了重要一步，而且还为其在临床诊断和治疗监测中的广泛应用奠定了基础。

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Innovative Colorimetric NQO1 Detection Strategy via Substrate Competitive and Biomimetic Cascade Reactions with a Highly Active NADH Oxidase Mimic

NAD(P)H: quinone oxidoreductase-1 (NQO1) plays critical roles in antioxidation and abnormally overexpresses in tumors. Developing a fast and sensitive method of monitoring NQO1 will greatly promote cancer diagnosis in clinical practice. This study introduces a transformative colorimetric detection strategy for NQO1, harnessing an innovative competitive substrate mechanism between NQO1 and a new NADH oxidase (NOX) mimic, cobalt–nitrogen-doped carbon nanozyme (CoNC). This method ingeniously exploits the differential consumption of NADH in the presence of NQO1 to modulate the generation of H₂O₂ from CoNC catalysis, which is then quantified through a secondary, peroxidase-mimetic cascade reaction involving Prussian blue (PB) nanoparticles. This dual-stage reaction framework not only enhances the sensitivity of NQO1 detection, achieving a limit of detection as low as 0.67 μg mL^–1, but also enables the differentiation between cancerous and noncancerous cells by their enzymatic activity profiles. Moreover, CoNC exhibits exceptional catalytic efficiency, with a specific activity reaching 5.2 U mg^–1, significantly outperforming existing NOX mimics. Beyond mere detection, CoNC serves a dual role, acting as both a robust mimic of cytochrome c reductase (Cyt c) and a cornerstone for enzymatic regeneration, thereby broadening the scope of its biological applications. This study not only marks a significant step forward in the bioanalytical application of nanozymes but also sets the stage for their expanded use in clinical diagnostics and therapeutic monitoring.

更新日期：2024-07-30

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