Noninvasive and Highly Selective Monitoring of Intracellular Glucose via a Two-Step Recognition-Based Nanokit
Noninvasive and Highly Selective Monitoring of Intracellular Glucose via a Two-Step Recognition-Based Nanokit
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通过基于两步识别的纳米试剂盒对细胞内葡萄糖进行无创和高选择性监测
DOI：
10.1021/acs.analchem.7b01532
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发表时间：
2017
影响因子：
7.4
通讯作者：
Yang Ronghua
中科院分区：
文献类型：
--
作者：
Tang Jianru;Ma D;an;Pecic Stevan;Huang Caixia;Zheng Jing;Li Jishan;Yang Ronghua
细胞内葡萄糖的准确测定对于探索其在活细胞代谢过程中的化学和生物学功能具有重要意义。在本文中，我们开发了一种新的非侵入性和高选择性的纳米试剂盒，通过两步识别的细胞内葡萄糖监测。基于脂质体的纳米试剂盒将适配体功能化的金纳米颗粒（AuNPs）和Shinkai的受体共同封装在一起。当将所提出的纳米试剂盒转染到活细胞中时，Shinkai的受体可以首先识别葡萄糖，然后改变其构象，赋予适体以结合和传感特性，这些特性在其他情况下不易获得。然后，由细胞内葡萄糖和Shinkai的受体形成的二元复合物可以原位置换AuNPs表面上的适体的互补寡核苷酸。荧光标记的适配体远离金纳米粒子，荧光状态从“关闭”切换到“打开”。通过适配体的二次鉴定，可以大大提高Shinkai受体的选择性，同时通过适配体的荧光信号恢复来评估细胞内葡萄糖水平。在后续的应用中，该方法表现出良好的选择性，是无创的细胞内葡萄糖监测在常氧和缺氧。据我们所知，这是第一次有机受体和核酸的优势相结合，并通过两步识别实现了对细胞内葡萄糖的高选择性监测。我们希望它能开辟新的可能性，将各种代谢疾病的诊断和胰岛素输送设备集成在一起。
Accurate determination of intracellular glucose is very important for exploring its chemical and biological functions in metabolism events of living cells. In this paper, we developed a new noninvasive and highly selective nanokit for intracellular glucose monitoring via two-step recognition. The liposome-based nanokit coencapsulated the aptamer-functionalized gold nanoparticles (AuNPs) and the Shinkai’s receptor together. When the proposed nanokit was transfected into living cells, the Shinkai’s receptor could recognize glucose first and then changed its conformation to endow aptamers with binding and sensing properties which were not readily accessible otherwise. Then, the binary complexes formed by the intracellular glucose and the Shinkai’s receptor canin situdisplace the complementary oligonucleotide of the aptamer on the surface of AuNPs. The fluorophore-labeled aptamer was away from the AuNPs, and the fluorescent state switched from “off” to “on”. Through the secondary identification of aptamer, the selectivity of the Shinkai’s receptor could be greatly improved while the intracellular glucose level was assessed by fluorescence signal recovery of aptamer. In the follow-up application, the approach exhibits excellent selectivity and is noninvasive for intracellular glucose monitoring under normoxia and hypoxia. To the best of our knowledge, this is the first time that the advantages of organic receptors and nucleic acids have been combined and highly selective monitoring of intracellular glucose has been realized via two-step recognition. We expect it to open up new possibilities to integrate devices for diagnosis of various metabolic diseases and insulin delivery.