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Single-walled carbon nanotubes (SWCNTs) emit photostable near-infrared fluorescence that is conducive for optical glucose monitoring. Such SWCNT-based optical sensors often require the immobilization of proteins that can confer glucose selectivity and reactivity. In this work, we immobilize a glucose-reactive enzyme, glucose oxidase, onto SWCNTs using a N (1 pyrenyl)maleimide (PM) crosslinker via thiol bioconjugation of engineered cysteine residues. We compare the conjugation of several glucose oxidase variants containing rationally-engineered cysteines and identify a D70C variant that shows effective bioconjugation. The bioconjugation was characterized through both absorption and fluorescence spectroscopy. Furthermore, we demonstrate an application for continuous glucose monitoring in the NIR-II optical region using the bioconjugated reaction solution, which shows a reversible response to physiological concentrations of glucose. Finally, we develop a miniaturized reader for monitoring the sensor response which will enable continuous glucose monitoring in cell cultures.
Christian Enz, Sandro Carrara, Assim Boukhayma, Antonino Caizzone, Ata Jedari Golparvar
Kei Sakamoto, Katyayanee Neopane