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基于生物矿化作用的集成免疫探针及其在免疫层析分析中的应用。

Biomineralization-powered integrated immunoprobe and its application in Immunochromatographic assay.

机构信息

State Key Laboratory of Food Science and Resources, Nanchang University, Nanchang, 330047, China.

College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, China.

出版信息

Biosens Bioelectron. 2024 Mar 15;248:115945. doi: 10.1016/j.bios.2023.115945. Epub 2023 Dec 20.

Abstract

Immunochromatographic assay (ICA) has attracted widespread attention owing to its advantages of economy, simplicity, and rapidity. However, the synthesis of immunoprobes is still limited by complicated design ideas and multistep operations from preparing nanoparticles to conjugating monoclonal antibodies (mAb) onto nanoparticles. Inspired by the biomineralization of zeolitic imidazolate framework-8 (ZIF-8), we proposed a strategy for the rapid synthesis of an integrated immunoprobe (ZIF-8@QDs-mAb), achieving a one-step integration with strong fluorescent signal output capability and specific recognition ability. In addition, different fluorescent colors of ZIF-8@QDs-mAb were generated by doping red and green quantum dots (QDs) in various ratios. With a smart detection platform, the developed ZIF-8@QDs-mAb-based multiplex ICA (ZIF-8@QDs-mAb-mICA) achieved the on-site quantitative detection of enrofloxacin, sulfamethazine, and kanamycin in milk within 15 min, with the limit of detection (LOD) of 0.052, 0.186 and 0.216 ng mL, which were 5.69, 2.20 and 4.40 times higher than that of gold nanoparticles-based mICA, respectively. The quantitative detection of alpha-fetoprotein and human chorionic gonadotropin was also achieved with LOD of 0.516 ng mL and 0.225 mIU mL, respectively, which verified the universality of the strategy. This work provides a novel idea for the design of an efficient integrated immunoprobe and has broad application prospects in ICA.

摘要

免疫胶体金分析(ICA)因其经济、简便、快速的优点而受到广泛关注。然而,免疫探针的合成仍然受到复杂的设计理念和从制备纳米粒子到将单克隆抗体(mAb)偶联到纳米粒子的多步操作的限制。受沸石咪唑酯骨架-8(ZIF-8)生物矿化的启发,我们提出了一种快速合成一体化免疫探针(ZIF-8@QDs-mAb)的策略,实现了具有强荧光信号输出能力和特异性识别能力的一步集成。此外,通过以不同比例掺杂红色和绿色量子点(QDs),生成了具有不同荧光颜色的 ZIF-8@QDs-mAb。通过智能检测平台,开发的基于 ZIF-8@QDs-mAb 的多重 ICA(ZIF-8@QDs-mAb-mICA)在 15 分钟内实现了牛奶中恩诺沙星、磺胺甲恶唑和卡那霉素的现场定量检测,检测限(LOD)分别为 0.052、0.186 和 0.216 ng mL,分别比基于金纳米粒子的 mICA 高 5.69、2.20 和 4.40 倍。对甲胎蛋白和人绒毛膜促性腺激素的定量检测也分别达到了 0.516 ng mL 和 0.225 mIU mL 的检测限,验证了该策略的通用性。这项工作为高效一体化免疫探针的设计提供了新的思路,并在 ICA 中具有广阔的应用前景。

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