Dong Yan, Dong Wei, Liang Xin, Wang Yuan-Rui, Xu Feng, Li Li, Han Lu, Jiang Li-Rui
College of Pharmacy, Qiqihar Medical University, Qiqihar 161006, PR China.
College of Pharmacy, Qiqihar Medical University, Qiqihar 161006, PR China.
Spectrochim Acta A Mol Biomol Spectrosc. 2023 May 15;293:122513. doi: 10.1016/j.saa.2023.122513. Epub 2023 Feb 16.
Thrombin (TB) plays a key role in the pathological and physiological coagulation of diseases. In this work, a TB-activated fluorescence-surface-enhanced Raman spectroscopy (SERS) dual-mode optical nanoprobe (MRAu) was constructed by linking rhodamine B (RB)-modified magnetic fluorescent nanospheres with AuNPs through TB-specific recognition peptides. In the presence of TB, the polypeptide substrate could specifically be cleaved by TB, resulting in the weakening of SERS hotspot effect and the reduction of Raman signal. Meanwhile, the fluorescence resonance energy transfer (FRET) system was destroyed, and the RB fluorescence signal originally quenched by AuNPs was recovered. Using MRAu, SERS and fluorescence methods were combined to extend the TB detection range to 1-150 pM, and the detection limit was as low as 0.35 pM. In addition, the ability to detect TB in human serum also verified the effectiveness and practicality of the nanoprobe. The probe was also successfully employed to evaluate the inhibitory effect against TB of active components in Panax notoginseng. This study provides a new technical means for the diagnosis and drug development of abnormal TB-related diseases.
凝血酶（TB）在疾病的病理和生理凝血过程中起着关键作用。在这项工作中，通过将罗丹明B（RB）修饰的磁性荧光纳米球与金纳米粒子（AuNPs）通过TB特异性识别肽连接，构建了一种TB激活的荧光-表面增强拉曼光谱（SERS）双模光学纳米探针（MRAu）。在TB存在的情况下，多肽底物可被TB特异性切割，导致SERS热点效应减弱和拉曼信号降低。同时，荧光共振能量转移（FRET）系统被破坏，原本被AuNPs淬灭的RB荧光信号得以恢复。利用MRAu，将SERS和荧光方法结合，将TB检测范围扩展到1-150 pM，检测限低至0.35 pM。此外，在人血清中检测TB的能力也验证了纳米探针的有效性和实用性。该探针还成功用于评估三七中活性成分对TB的抑制作用。本研究为TB相关异常疾病的诊断和药物开发提供了一种新的技术手段。
