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基于特异性结合生物受体和碳量子点的组胺荧光检测。

Fluorescence detection of histamine based on specific binding bioreceptors and carbon quantum dots.

机构信息

Department of Chemistry, Research Institute of Chem-Bio Diagnostic Technology, Chung-Ang University, 84 Heukseok-ro, Dongjak-gu, Seoul 06974, Republic of Korea.

Department of Food Science and Biotechnology, School of Food Science and Technology, Chung-Ang University, 4726 Seodong-daero, Anseong 17546, Republic of Korea.

出版信息

Biosens Bioelectron. 2020 Nov 1;167:112519. doi: 10.1016/j.bios.2020.112519. Epub 2020 Aug 14.

Abstract

Histamine is primarily found in spoiled food and often used as an indicator of food safety. Compared to various existing methods for analyzing histamine, high-performance liquid chromatography (HPLC) which is accurate but time-consuming, and immunochemical methods that are difficult to produce high specificity and affinity antibodies towards small molecules have been used. In this study, we developed a newly designed, sensitive, and selective fluorescence detection platform for histamine sensing, utilizing carbon quantum dots (CQDs) and synthetic peptides. Specifically, through biopanning approaches, a series of peptides having a high affinity towards immobilized histamine hapten were selected from phage-displayed libraries. Then, CQDs were synthesized by one-pot hydrothermal treatment enabling their fluorescence to be effectively quenched by peptides via the electron transfer interactions. While, in the presence of histamine, fluorescence will be recovered because of the stronger interaction between peptide and target. In this study, from the selectivity tests towards histamine and in contrast to structurally similar compounds, peptide Hisp3 (DIDRAGKASHWP) along with its dipeptide repeat derivative (Hisp3-2-C) were chemically synthesized to be used as promising histamine receptors. Furthermore, the application of peptide along with gold-coated magnetic nanoparticles (MNP@Au NPs) was designed for purification and analysis of fish samples. These results indicate that the CQDs and peptide sensor system could detect histamine at lower concentrations with high sensitivity and selectivity.

摘要

组氨酸主要存在于变质的食物中,常被用作食品安全的指标。与各种现有的组胺分析方法相比,高效液相色谱(HPLC)虽然准确但耗时,免疫化学方法难以产生针对小分子的高特异性和亲和力抗体。在这项研究中,我们开发了一种新设计的、灵敏且选择性的荧光检测平台,用于组胺传感,利用碳量子点(CQDs)和合成肽。具体来说,通过生物淘选方法,从噬菌体展示文库中筛选出一系列对固定化组氨酸半抗原具有高亲和力的肽。然后,通过一锅水热处理合成 CQDs,使它们的荧光能够通过电子转移相互作用有效地被肽猝灭。然而,在存在组胺的情况下,由于肽与靶标之间更强的相互作用,荧光将恢复。在这项研究中,从对组胺的选择性测试来看,与结构相似的化合物相比,肽 Hisp3(DIDRAGKASHWP)及其二肽重复衍生物(Hisp3-2-C)被化学合成,用作有前途的组氨酸受体。此外,肽与金涂覆的磁性纳米颗粒(MNP@Au NPs)的应用被设计用于鱼样的纯化和分析。这些结果表明,CQDs 和肽传感器系统可以在较低浓度下以高灵敏度和选择性检测组胺。

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