Engineered Glucose Oxidase-Carbon Nanotube Conjugates for Tissue-Translatable Glucose Nanosensors

Continuous and non-invasive glucose monitoring and imaging is important for disease diagnosis, treatment, and management. However, glucose monitoring remains a technical challenge owing to the dearth of tissue-transparent glucose sensors. In this study, we present the development of near-infrared fluorescent single-walled carbon nanotube (SWCNT) based nanosensors directly functionalized with glucose oxidase (GOx) capable of immediate and reversible glucose imaging in biological fluids and tissues. We prepared GOx-SWCNT nanosensors by facile sonication of SWCNT with GOx in a manner that-surprisingly-does not compromise the ability of GOx to detect glucose. Importantly, we find by using denatured GOx that the fluorescence modulation of GOx-SWCNT is not associated with the catalytic oxidation of glucose but rather triggered by glucose-GOx binding. Leveraging the unique response mechanism of GOx-SWCNT nanosensors, we developed catalytically inactive apo-GOx-SWCNT that enables both sensitive and reversible glucose imaging, exhibiting a ΔF/F₀ of up to 40 % within 1 s of exposure to glucose without consuming the glucose analyte. We finally demonstrate the potential applicability of apo-GOx-SWCNT in biomedical applications by glucose quantification in human plasma and glucose imaging in mouse brain slices.