Rationally Designed Functionalization of Single‐Walled Carbon Nanotubes for Real‐Time Monitoring of Cholinesterase Activity and Inhibition in Plasma

Abstract Enzymes play a pivotal role in regulating numerous bodily functions. Thus, there is a growing need for developing sensors enabling real‐time monitoring of enzymatic activity and inhibition. The activity and inhibition of cholinesterase (CHE) enzymes in blood plasma are fluorometrically monitored using near‐infrared (NIR) fluorescent single‐walled carbon nanotubes (SWCNTs) as probes, strategically functionalized with myristoylcholine (MC)– the substrate of CHE. A significant decrease in the fluorescence intensity of MC‐suspended SWCNTs upon interaction with CHE is observed, attributed to the hydrolysis of the MC corona phase of the SWCNTs by CHE. Complementary measurements for quantifying choline, the product of MC hydrolysis, reveal a correlation between the fluorescence intensity decrease and the amount of released choline, rendering the SWCNTs optical sensors with real‐time feedback in the NIR biologically transparent spectral range. Moreover, when synthetic and naturally abundant inhibitors inhibit the CHE enzymes present in blood plasma, no significant modulations of the MC‐SWCNT fluorescence are observed, allowing effective detection of CHE inhibition. The rationally designed SWCNT sensors platform for monitoring of enzymatic activity and inhibition in clinically relevant samples is envisioned to not only advance the field of clinical diagnostics but also deepen further understanding of enzyme‐related processes in complex biological fluids.