Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China.
Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China; Shanghai Key Laboratory of Bio-Energy Crops, Shanghai University, Shanghai 200444, China.
Biosens Bioelectron. 2016 Apr 15;78:100-105. doi: 10.1016/j.bios.2015.11.034. Epub 2015 Nov 14.
Herein, a binding-regulated click ligation (BRCL) strategy for endowing selective detection of proteins is developed with the incorporation of small-molecule ligand and clickable DNA probes. The fundamental principle underlying the strategy is the regulating capability of specific protein-ligand binding against the ligation between clickable DNA probes, which could efficiently combine the detection of particular protein with enormous DNA-based sensing technologies. In this work, the feasibly of the BRCL strategy is first verified through agarose gel electrophoresis and electrochemical impedance spectroscopy measurements, and then confirmed by transferring it to a nanomaterial-assisted fluorescence assay. Significantly, the BRCL strategy-based assay is able to respond to target protein with desirable selectivity, attributing to the specific recognition between small-molecule ligand and its target. Further experiments validate the general applicability of the sensing method by tailoring the ligand toward different proteins (i.e., avidin and folate receptor), and demonstrate its usability in complex biological samples. To our knowledge, this work pioneers the practice of click chemistry in probing specific small-molecule ligand-protein binding, and therefore may pave a new way for selective detection of proteins.
本文提出了一种结合小分子配体和可点击 DNA 探针的结合调控点击连接（BRCL）策略，用于赋予蛋白质选择性检测能力。该策略的基本原理是特定蛋白-配体结合对可点击 DNA 探针之间连接的调控能力，这可以有效地将特定蛋白质的检测与大量基于 DNA 的传感技术结合起来。在这项工作中，首先通过琼脂糖凝胶电泳和电化学阻抗谱测量验证了 BRCL 策略的可行性，然后通过将其转化为纳米材料辅助荧光分析进一步得到证实。值得注意的是，基于 BRCL 策略的分析能够对目标蛋白进行理想的选择性响应，这归因于小分子配体与其靶蛋白之间的特异性识别。进一步的实验通过将配体定制为不同的蛋白质（即亲和素和叶酸受体），验证了该传感方法的通用性，并证明了其在复杂生物样品中的可用性。据我们所知，这项工作开创了点击化学在探测特定小分子配体-蛋白质结合方面的应用，因此可能为蛋白质的选择性检测开辟新途径。
