Determination of intracellular esterase activity using ratiometric Raman sensing and spectral phasor analysis

Braddick, Henry J. and Tipping, William J. and Wilson, Liam T. and Jaconelli, Harry S. and Grant, Emma K. and Faulds, Karen and Graham, Duncan and Tomkinson, Nicholas C. O. (2023) Determination of intracellular esterase activity using ratiometric Raman sensing and spectral phasor analysis. Analytical Chemistry, 95 (12). 5369–5376. ISSN 0003-2700 (https://doi.org/10.1021/acs.analchem.2c05708)

[thumbnail of Braddick-etal-AC-2023-Determination-of-intracellular-esterase-activity-using-ratiometric-Raman-sensing]
Preview
 Text. Filename: Braddick_etal_AC_2023_Determination_of_intracellular_esterase_activity_using_ratiometric_Raman_sensing.pdf
Final Published Version
License: Creative Commons Attribution 4.0 logo
Download (4MB)| Preview

Abstract

Carboxylesterases (CEs) are a class of enzyme that catalyse 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 UV damage in a mixed cell population was possible using stimulated Raman scattering (SRS) 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.

CORE (COnnecting REpositories)