Department of Pure and Applied Chemistry, Thomas Graham Building, University of Strathclyde, 295 Cathedral Street, Glasgow G1 1XL, U.K.
Centre for Molecular Nanometrology, Department of Pure and Applied Chemistry, Technology and Innovation Centre, University of Strathclyde, 99 George Street, Glasgow G1 1RD, U.K.
Anal Chem. 2023 Mar 28;95(12):5369-5376. doi: 10.1021/acs.analchem.2c05708. Epub 2023 Mar 16.
Carboxylesterases (CEs) are a class of enzymes that catalyze the hydrolysis of esters in a variety of endogenous and exogenous molecules. CEs play an important role in drug metabolism, in the onset and progression of disease, and can be harnessed for prodrug activation strategies. As such, the regulation of CEs is an important clinical and pharmaceutical consideration. Here, we report the first ratiometric sensor for CE activity using Raman spectroscopy based on a bisarylbutadiyne scaffold. The sensor was shown to be highly sensitive and specific for CE detection and had low cellular cytotoxicity. In hepatocyte cells, the ratiometric detection of esterase activity was possible, and the result was validated by multimodal imaging with standard viability stains used for fluorescence microscopy within the same cell population. In addition, we show that the detection of localized ultraviolet damage in a mixed cell population was possible using stimulated Raman scattering microscopy coupled with spectral phasor analysis. This sensor demonstrates the practical advantages of low molecular weight sensors that are detected using ratiometric Raman imaging and will have applications in drug discovery and biomedical research.
羧酸酯酶（Carboxylesterases，CEs）是一类能够催化多种内源性和外源性分子中酯键水解的酶。CEs 在药物代谢、疾病的发生和进展中发挥着重要作用，并且可以被用于前药激活策略。因此，CE 的调控是临床和药物学方面的重要考虑因素。在这里，我们报道了首个基于双芳基丁二炔支架的比率型 CE 活性拉曼光谱传感器。该传感器对 CE 检测具有高度的灵敏性和特异性，并且细胞毒性低。在肝细胞中，可以实现酯酶活性的比率型检测，并且通过与同一细胞群体中荧光显微镜中使用的标准活力染色剂相结合的多模态成像对结果进行了验证。此外，我们还表明，使用结合了光谱相分析的受激拉曼散射显微镜，可以检测混合细胞群体中的局部紫外损伤。该传感器展示了使用比率型拉曼成像检测的低分子量传感器的实际优势，将在药物发现和生物医学研究中得到应用。
