Bartolini Cosimo, Tozzetti Martina, Gellini Cristina, Ricci Marilena, Menichetti Stefano, Procacci Piero, Caminati Gabriella
Department of Chemistry "Ugo Schiff", University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, FI, Italy.
Center for Colloid and Surface Science (CSGI), University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, FI, Italy.
Nanomaterials (Basel). 2025 Aug 12;15(16):1230. doi: 10.3390/nano15161230.
We present a dual biosensing strategy integrating Quartz Crystal Microbalance (QCM) and Surface-Enhanced Raman Spectroscopy (SERS) for the quantitative and molecular-specific detection of FKBP12. Silver nanodendritic arrays were electrodeposited onto QCM sensors, optimized for SERS enhancement using Rhodamine 6G, and functionalized with a custom-designed receptor to selectively capture FKBP12. QCM measurements revealed a two-step Langmuir adsorption behavior, enabling sensitive mass quantification with a low limit of detection. Concurrently, in situ SERS analysis on the same sensor provided vibrational fingerprints of FKBP12, resolved through comparative studies of the free protein, surface-bound receptor, and surface-bound receptor-protein complex. Ethanol-induced denaturation confirmed protein-specific peaks, while shifts in receptor vibrational modes-linked to FKBP12 binding-demonstrated dynamic molecular interactions. A ratiometric parameter, derived from key peak intensities, served as a robust, concentration-dependent signature of complex formation. This platform bridges quantitative (QCM) and structural (SERS) biosensing, offering real-time mass tracking and conformational insights. The nanodendritic substrate's dual functionality, combined with the receptor's selectivity, advances label-free protein detection for applications in drug diagnostics, with potential adaptability to other target analytes.
我们提出了一种将石英晶体微天平(QCM)和表面增强拉曼光谱(SERS)相结合的双生物传感策略,用于对FKBP12进行定量和分子特异性检测。将银纳米树枝状阵列电沉积到QCM传感器上,使用罗丹明6G对其进行优化以增强SERS,并使用定制设计的受体进行功能化,以选择性捕获FKBP12。QCM测量揭示了两步朗缪尔吸附行为,能够以低检测限进行灵敏的质量定量。同时,在同一传感器上进行的原位SERS分析提供了FKBP12的振动指纹图谱,通过对游离蛋白质、表面结合受体和表面结合受体-蛋白质复合物的比较研究得以解析。乙醇诱导的变性证实了蛋白质特异性峰,而与FKBP12结合相关的受体振动模式的变化表明了动态分子相互作用。由关键峰强度得出的比例参数作为复合物形成的稳健、浓度依赖性特征。该平台将定量(QCM)和结构(SERS)生物传感相结合,提供实时质量跟踪和构象洞察。纳米树枝状基底的双重功能与受体的选择性相结合,推动了无标记蛋白质检测在药物诊断中的应用,并有可能适用于其他目标分析物。