Metal–Organic Framework-Based Cascade Catalysis-Enabled Fluorescent ELISA with Higher Enzymatic Stability for Zearalenone Detection in Maize

IF 5.7 1区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY
Chaochao Chen, Liang Luo, Jianzhong Shen, Zhanhui Wang, Yantong Pan
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Abstract

The enzyme-linked immunosorbent assay (ELISA) stands as one of the most frequently employed rapid detection techniques for both chemical and biological contaminants. Horseradish peroxidase (HRP) and hydrogen peroxide (H2O2), serving as signal generators, play indispensable roles throughout the entire ELISA process. However, HRP and H2O2 exhibit high sensitivity to elevated temperatures, hindering the broader utilization and transportation of ELISA. Hereby, being inspired by the cascade catalysis of glucose by glucose oxidase (GOx) and HRP as well as the advantageous properties of metal–organic frameworks (MOFs), we developed a cascade catalysis-based competitive fluorescent ELISA (CCF-ELISA) for mycotoxin detection in maize using zearalenone (ZEN) as a model. The results revealed the remarkable protective effects of MOFs on enzymatic activities. Based on upon characteristics, the built CCF-ELISA enabled the detection of ZEN with a half-maximal inhibitory concentration of 0.54 ng/mL and allowed a detection range from 0.19 to 1.51 ng/mL. When applied to naturally contaminated maize samples, the outcomes obtained from the CCF-ELISA showed a high level of comparability with high-performance liquid chromatography–tandem mass spectrometry, demonstrating the promising potential of the developed CCF-ELISA in the rapid detection for food security.

Abstract Image

金属-有机框架级联催化荧光酶联免疫吸附法检测玉米玉米赤霉烯酮
酶联免疫吸附试验(ELISA)是化学和生物污染物最常用的快速检测技术之一。辣根过氧化物酶(HRP)和过氧化氢(H2O2)作为信号发生器,在整个ELISA过程中起着不可或缺的作用。然而,HRP和H2O2对高温敏感,阻碍了ELISA的广泛利用和运输。因此,受葡萄糖氧化酶(GOx)和HRP对葡萄糖的级联催化作用的启发,以及金属有机框架(mof)的优势特性,我们以玉米赤霉烯酮(ZEN)为模型,开发了一种基于级联催化的竞争性荧光ELISA (CCF-ELISA)检测玉米真菌毒素。结果表明,mof对酶活性具有显著的保护作用。基于以上特点,构建的CCF-ELISA检测ZEN的半数最大抑制浓度为0.54 ng/mL,检测范围为0.19 ~ 1.51 ng/mL。CCF-ELISA与高效液相色谱-串联质谱分析结果具有较高的可比性,证明了CCF-ELISA在食品安全快速检测中的应用前景。
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来源期刊
Journal of Agricultural and Food Chemistry
Journal of Agricultural and Food Chemistry 农林科学-农业综合
CiteScore
9.90
自引率
8.20%
发文量
1375
审稿时长
2.3 months
期刊介绍: The Journal of Agricultural and Food Chemistry publishes high-quality, cutting edge original research representing complete studies and research advances dealing with the chemistry and biochemistry of agriculture and food. The Journal also encourages papers with chemistry and/or biochemistry as a major component combined with biological/sensory/nutritional/toxicological evaluation related to agriculture and/or food.
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阿拉丁
fluorescein isothiocyanate isomer I (FITC)
阿拉丁
rhodamine B isothiocyanate (RhB)
阿拉丁
fluorescein isothiocyanate isomer I (FITC)
阿拉丁
Rhodamine B isothiocyanate (RhB)
Sigma
ochratoxin A (OTA)
Sigma
aflatoxin B1 (AFB1)
Sigma
fumonisin B1 (FB1)
Sigma
deoxynivalenol (DON)
Sigma
zinc nitrate hexahydrate (Zn(NO3)2·6H2O)
Sigma
GOx
Sigma
glucose
Sigma
bovine serum albumin (BSA)
Sigma
ethylenediamine tetraacetic acid (EDTA)
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