Bioengineering a glucose oxidase nanosensor for near-infrared continuous glucose monitoring

Vitalijs Zubkovs; Hanxuan Wang; Nils Schuergers; Astrid Weninger; Anton Glieder; Stefano Cattaneo; Ardemis A. Boghossian

doi:10.1039/d2na00092j

Bioengineering a glucose oxidase nanosensor for near-infrared continuous glucose monitoring

Vitalijs Zubkovs, Hanxuan Wang, Nils Schuergers, Astrid Weninger, Anton Glieder, Stefano Cattaneo, Ardemis A. Boghossian^*

^*Corresponding author for this work

Institute of Molecular Biotechnology (6550)

Research output: Contribution to journal › Article › peer-review

Abstract

Single-walled carbon nanotubes (SWCNTs) emit photostable near-infrared (NIR) 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 (GOx), 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 NIR-II reader to be used for cell cultures that require continuous glucose monitoring.

Original language	English
Pages (from-to)	2420-2427
Number of pages	8
Journal	Nanoscale Advances
Volume	4
Issue number	11
DOIs	https://doi.org/10.1039/d2na00092j
Publication status	Published - 4 May 2022

ASJC Scopus subject areas

Bioengineering
Atomic and Molecular Physics, and Optics
Chemistry(all)
Materials Science(all)
Engineering(all)

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title = "Bioengineering a glucose oxidase nanosensor for near-infrared continuous glucose monitoring",

abstract = "Single-walled carbon nanotubes (SWCNTs) emit photostable near-infrared (NIR) 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 (GOx), 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 NIR-II reader to be used for cell cultures that require continuous glucose monitoring.",

author = "Vitalijs Zubkovs and Hanxuan Wang and Nils Schuergers and Astrid Weninger and Anton Glieder and Stefano Cattaneo and Boghossian, {Ardemis A.}",

note = "Funding Information: The authors are thankful for support from the Swiss National Science Foundation Assistant Professor (AP) Energy Grant (Project No. PYAPP2_154269) and Swiss National Science Foundation Project No. 200021_184822. This project has received funding from the European Research Council (ERC) under the European Union{\textquoteright}s Horizon 2020 research and innovation programme (grant agreement No 853005). The authors acknowledge E. Ahunbay and M. Moradel-Casellas, who helped establish protocols for glucose oxidase expression and purification, as well as S. Y. Rahnamaee, who helped to express and purify the proteins. G. Orawez developed the electronics design of the miniaturized device and its firmware, while T. Tatar developed the device software. E. Vuille-Dit-Bille performed sensor monitoring experiments. Publisher Copyright: {\textcopyright} 2022 RSC",

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AU - Zubkovs, Vitalijs

AU - Wang, Hanxuan

AU - Schuergers, Nils

AU - Weninger, Astrid

AU - Glieder, Anton

AU - Cattaneo, Stefano

AU - Boghossian, Ardemis A.

N1 - Funding Information: The authors are thankful for support from the Swiss National Science Foundation Assistant Professor (AP) Energy Grant (Project No. PYAPP2_154269) and Swiss National Science Foundation Project No. 200021_184822. This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No 853005). The authors acknowledge E. Ahunbay and M. Moradel-Casellas, who helped establish protocols for glucose oxidase expression and purification, as well as S. Y. Rahnamaee, who helped to express and purify the proteins. G. Orawez developed the electronics design of the miniaturized device and its firmware, while T. Tatar developed the device software. E. Vuille-Dit-Bille performed sensor monitoring experiments. Publisher Copyright: © 2022 RSC

PY - 2022/5/4

Y1 - 2022/5/4

N2 - Single-walled carbon nanotubes (SWCNTs) emit photostable near-infrared (NIR) 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 (GOx), 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 NIR-II reader to be used for cell cultures that require continuous glucose monitoring.

AB - Single-walled carbon nanotubes (SWCNTs) emit photostable near-infrared (NIR) 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 (GOx), 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 NIR-II reader to be used for cell cultures that require continuous glucose monitoring.

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Bioengineering a glucose oxidase nanosensor for near-infrared continuous glucose monitoring

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