基于碱性磷酸酶介导的纳米银颗粒生长和荧光内滤效应的比色-荧光双模纳米传感器驱动酶免疫测定赭曲霉毒素A

IF 11.3 1区环境科学与生态学 Q1 ENGINEERING, ENVIRONMENTAL

Journal of Hazardous Materials Pub Date : 2025-05-10 DOI:10.1016/j.jhazmat.2025.138539

Xincheng Chen, Zhenyun He, Sujia Jiao, Zhichang Sun, Sihang Zhang, Xing Liu

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引用次数: 0

摘要

赭曲霉毒素A （Ochratoxin A， OTA）严重威胁公众健康和农业生态系统，需要灵敏的检测方法。鉴于传统的免疫检测依赖于动物源性抗体和有毒底物，我们开发了一种利用碱性磷酸酶（ALP）介导的银纳米颗粒（AgNPs）生长和内部过滤效应（IFE）集成比色和荧光检测的双模式纳米传感器。为了构建双模纳米传感器，研究了银离子触发金纳米团簇（aunc）的聚集诱导发射（AIE）增强。该纳米传感器采用ota特异性纳米体- alp融合（Nb-ALP）将抗坏血酸2-磷酸去磷酸化为抗坏血酸（AA），从而将Ag +还原为含有nabh4还原AgNPs种子的丰富AgNPs。AgNPs放大等离子体吸光度用于比色ALP检测，同时通过IFE猝灭Ag + @AuNC荧光用于荧光检测。该纳米传感器进一步与基于nb - alp的免疫分析相结合，开发了一种双模式纳米传感器驱动的酶免疫分析（DMN-EIA）。DMN-EIA对OTA的检出限分别为0.14 ng/mL（比色法）和0.28 ng/mL（荧光法），具有较高的选择性，峰回收率为93.5% ~ 108.7%，相对标准偏差不超过20%。比色法（R2 = 0.96）和荧光DMN-EIA法（R2 = 0.93）与高效液相色谱法检测10种辣椒样品具有良好的相关性。因此，所开发的双模纳米传感器和DMN-EIA代表了检测OTA的可靠和有前途的工具。

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Colorimetric-fluorescent dual-mode nanosensor-powered enzyme immunoassay for ochratoxin A via alkaline phosphatase-mediated silver nanoparticle growth and fluorescence inner filter effect

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Colorimetric-fluorescent dual-mode nanosensor-powered enzyme immunoassay for ochratoxin A via alkaline phosphatase-mediated silver nanoparticle growth and fluorescence inner filter effect

Ochratoxin A (OTA) seriously threatens public health and agricultural ecosystems, necessitating sensitive detection methods. In view of that traditional immunoassays rely on animal-derived antibodies and toxic substrates, we developed a dual-mode nanosensor integrating colorimetric and fluorescent detection via alkaline phosphatase (ALP)-mediated silver nanoparticles (AgNPs) growth and inner filter effect (IFE). The Ag⁺-triggered aggregation-induced emission (AIE) enhancement of gold nanoclusters (AuNCs) was investigated for constructing a dual-mode nanosensor. The nanosensor employs an OTA-specific nanobody-ALP fusion (Nb-ALP) to dephosphorylate ascorbic acid 2-phosphate into ascorbic acid (AA), which reduces Ag⁺ to abundant AgNPs with NaBH₄-reduced AgNPs seeds. AgNPs amplify plasmonic absorbance for colorimetric ALP detection, while quenching Ag⁺@AuNC fluorescence via IFE for fluorescent detection. The nanosensor was further combined with the Nb-ALP-based immunoassay to develop a dual-mode nanosensor-powered enzyme immunoassay (DMN-EIA). The DMN-EIA exhibited detection limits of 0.14 ng/mL (colorimetric) and 0.28 ng/mL (fluorescent) with high selectivity for OTA, with spike recovery rates of 93.5%–108.7% and relative standard deviations not exceeding 20%. Furthermore, the colorimetric (R² = 0.96) and fluorescent DMN-EIA (R² = 0.93) correlated well with high-performance liquid chromatography in detection of ten real pepper samples. Therefore, the developed dual-mode nanosensor and DMN-EIA represent reliable and promising tools for detecting OTA.

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来源期刊

Journal of Hazardous Materials 工程技术-工程：环境

CiteScore

25.40

自引率

5.90%

发文量

3059

审稿时长

58 days

期刊介绍： The Journal of Hazardous Materials serves as a global platform for promoting cutting-edge research in the field of Environmental Science and Engineering. Our publication features a wide range of articles, including full-length research papers, review articles, and perspectives, with the aim of enhancing our understanding of the dangers and risks associated with various materials concerning public health and the environment. It is important to note that the term "environmental contaminants" refers specifically to substances that pose hazardous effects through contamination, while excluding those that do not have such impacts on the environment or human health. Moreover, we emphasize the distinction between wastes and hazardous materials in order to provide further clarity on the scope of the journal. We have a keen interest in exploring specific compounds and microbial agents that have adverse effects on the environment.