Xu Xinqiu, Jia Chaofan, Zhang Fengjiao, Li Hao, Gong Weilei, Wang Changqin, Wei Yin
School of Public Health, Shandong First Medical University and Shandong Academy of Medical Sciences, Jinan, Shandong, China.
School of Public Health, Jining Medical University, Jining, China.
J Fluoresc. 2025 Mar 28. doi: 10.1007/s10895-025-04282-2.
This study presents a rapid, simple aptamer-based fluorescence sensor using fullerenes as a quencher for detecting vitamin D3. Fullerenes, with its π-electron cloud, acts as a powerful electron acceptor, facilitating Förster resonance energy transfer (FRET) from carboxyfluorescein (FAM). The aptamer, labeled with 5'6-FAM, adsorbs onto the fullerenes surface through hydrogen bonding and π-π stacking interactions, leading to fluorescence quenching. Upon vitamin D3 binding, the aptamer forms a hairpin structure that prevents adsorption onto the fullerenes surface, restoring fluorescence. The fullerenes were characterized using UV-Vis, FT-IR, Raman spectroscopy, SEM, TEM, and zeta potential measurements. The sensor's response to varying vitamin D3 concentrations was analyzed with a fluorescence spectrometer, revealing a linear detection range of 0-600 nM and a detection limit of 200 nM, which improved to 50 nM with exonuclease I. The sensor demonstrated a recovery rate of 88.4%-96.3% for vitamin D3 in water samples, confirming its feasibility for practical applications. In addition, our proposed sensor exhibited good repeatability (3.27%) and stability (90%). The innovative use of fullerenes as a fluorescence quencher, along with the aptamer's specific binding to vitamin D3, provides a novel and highly sensitive avenue for vitamin D3 detection.
本研究提出了一种基于适体的快速、简单的荧光传感器，该传感器使用富勒烯作为猝灭剂来检测维生素D3。富勒烯具有π电子云，可作为强大的电子受体，促进羧基荧光素（FAM）的荧光共振能量转移（FRET）。用5'6-FAM标记的适体通过氢键和π-π堆积相互作用吸附在富勒烯表面，导致荧光猝灭。当维生素D3结合时，适体形成发夹结构，阻止其吸附在富勒烯表面，从而恢复荧光。使用紫外可见光谱、傅里叶变换红外光谱、拉曼光谱、扫描电子显微镜、透射电子显微镜和zeta电位测量对富勒烯进行了表征。用荧光光谱仪分析了该传感器对不同浓度维生素D3的响应，结果表明其线性检测范围为0-600 nM，检测限为200 nM，用核酸外切酶I处理后检测限提高到50 nM。该传感器对水样中维生素D3的回收率为88.4%-96.3%，证实了其实际应用的可行性。此外，我们提出的传感器具有良好的重复性（3.27%）和稳定性（90%）。富勒烯作为荧光猝灭剂的创新应用，以及适体与维生素D3的特异性结合，为维生素D3的检测提供了一条新颖且高度灵敏的途径。
