A novel fluorescent sensing platform for insulin detection based on competitive recognition of cationic pillar[6]arene
A novel fluorescent sensing platform for insulin detection based on competitive recognition of cationic pillar[6]arene
复制标题
基于阳离子柱[6]芳烃竞争识别的新型胰岛素检测荧光传感平台
DOI：
10.1016/j.talanta.2019.01.004
复制
发表时间：
2019-05-15
期刊：
影响因子：
6.1
通讯作者：
Li, Can-Peng
中科院分区：
文献类型：
--
作者：
Tan, Shuang;Han, Rui;Li, Can-Peng
竞争主客体识别已被用于利用大环超分子宿主来确定小分子，而较少的研究集中在蛋白质的特异性识别和传感上。在本工作中，我们首次报道了一种基于阳离子柱[6]芳烃(CP6)与胰岛素的超分子识别的无标记荧光测定胰岛素的方法。该方法基于CP6官能化还原氧化石墨烯(CP6@rGO)与探针/胰岛素分子竞争识别的荧光共振能量转移(FRET)。探针分子(RhB)具有很强的荧光信号，其荧光被基于FRET的rGO猝灭。当靶蛋白分子(胰岛素)被加入到CP6@rGO中时，探针被胰岛素取代，形成主客体复合体CP6@rGO/胰岛素，从而产生一个“开启”的荧光信号。络合物的荧光强度分别在0.01-0.50和1.0-16.0 mU M范围内与胰岛素浓度呈线性关系，检出限为3 nM。该传感器已成功应用于人工血清中胰岛素的测定。分子对接结果表明，胰岛素B链的N-末端Phe通过静电作用进入CP6空腔，形成稳定的主客体络合物。
Competitive host-guest recognition has been utilized to determine small molecules using macrocyclic supra molecular host, while less studies focused on the specific recognition and sensing of protein. In the present work, we are the first time to report a label-free fluorescent assay for insulin determination based on the supramolecular recognition between cationic pillar[6]arene (CP6) and insulin. The approach is based on fluorescence resonance energy transfer (FRET) through competitive recognition between CP6 functionalized reduced graphene oxide (CP6@rGO) and probe/insulin molecules. Probe molecule (RhB) has strong fluorescent signal, and its fluorescent is quenched by rGO based on FRET. When target protein molecule (insulin) is added to CP6@rGO, the probe is displaced by insulin and a host-guest complex CP6@rGO/insulin is formed, resulting in a "turn-on" fluorescence signal. The fluorescence intensity of complex increased linearly with the increase of insulin concentration ranging 0.01-0.50 and 1.0-16.0 mu M, respectively with a detection limit of 3 nM. The sensor was successfully utilized to determine insulin in artificial serum. The molecular docking result showed that the N-terminal Phe of insulin's B chain was included in the CP6 cavity through electrostatic interaction and formed a stable host-guest complex.