ConA-based glucose sensing using the long-lifetime azadioxatriangulenium fluorophore

Brian Cummins, Jonathan Simpson, Zygmunt Gryczynski, Thomas Just Sørensen, Bo W. Laursen, Duncan Graham, David Birch, Gerard Coté

Research output: Chapter in Book/Report/Conference proceedingConference contributionResearchpeer-review

2 Citations (Scopus)

Abstract

Fluorescent glucose sensing technologies have been identified as possible alternatives to current continuous glucose monitoring approaches. We have recently introduced a new, smart fluorescent ligand to overcome the traditional problems of ConA-based glucose sensors. For this assay to be translated into a continuous glucose monitoring device where both components are free in solution, the molecular weight of the smart fluorescent ligand must be increased. We have identified ovalbumin as a naturally-occurring glycoprotein that could serve as the core-component of a 2nd generation smart fluorescent ligand. It has a single asparagine residue that is capable of displaying an N-linked glycan and a similar isoelectric point to ConA. Thus, binding between ConA and ovalbumin can potentially be monovalent and sugar specific. This work is the preliminary implementation of fluorescently-labeled ovalbumin in the ConA-based assay. We conjugate the red-emitting, long-lifetime azadioxatriangulenium (ADOTA+) dye to ovalbumin, as ADOTA have many advantageous properties to track the equilibrium binding of the assay. The ADOTA-labeled ovalbumin is paired with Alexa Fluor 647-labeled ConA to create a Förster Resonance Energy Transfer (FRET) assay that is glucose dependent. The assay responds across the physiologically relevant glucose range (0-500 mg/dL) with increasing intensity from the ADOTA-ovalbumin, showing that the strategy may allow for the translation of the smart fluorescent ligand concept into a continuous glucose monitoring device.

Original languageEnglish
Title of host publicationOptical Diagnostics and Sensing XIV
Subtitle of host publicationToward Point-of-Care Diagnostics
PublisherSPIE
ISBN (Print)9780819498649
DOIs
StatePublished - 1 Jan 2014
EventOptical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics - San Francisco, CA, United States
Duration: 3 Feb 20146 Feb 2014

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume8951
ISSN (Print)1605-7422

Other

OtherOptical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics
CountryUnited States
CitySan Francisco, CA
Period3/02/146/02/14

Fingerprint

Fluorophores
Ovalbumin
glucose
Glucose
Assays
life (durability)
Ligands
ligands
Monitoring
Glucose sensors
Glycoproteins
Asparagine
Equipment and Supplies
Sugars
Energy Transfer
Isoelectric Point
Energy transfer
Polysaccharides
sugars
Coloring Agents

Keywords

  • Glucose sensing
  • competitive binding
  • energy transfer
  • fluorescence

Cite this

Cummins, B., Simpson, J., Gryczynski, Z., Sørensen, T. J., Laursen, B. W., Graham, D., ... Coté, G. (2014). ConA-based glucose sensing using the long-lifetime azadioxatriangulenium fluorophore. In Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics [89510A] (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 8951). SPIE. https://doi.org/10.1117/12.2039824
Cummins, Brian ; Simpson, Jonathan ; Gryczynski, Zygmunt ; Sørensen, Thomas Just ; Laursen, Bo W. ; Graham, Duncan ; Birch, David ; Coté, Gerard. / ConA-based glucose sensing using the long-lifetime azadioxatriangulenium fluorophore. Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics. SPIE, 2014. (Progress in Biomedical Optics and Imaging - Proceedings of SPIE).
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abstract = "Fluorescent glucose sensing technologies have been identified as possible alternatives to current continuous glucose monitoring approaches. We have recently introduced a new, smart fluorescent ligand to overcome the traditional problems of ConA-based glucose sensors. For this assay to be translated into a continuous glucose monitoring device where both components are free in solution, the molecular weight of the smart fluorescent ligand must be increased. We have identified ovalbumin as a naturally-occurring glycoprotein that could serve as the core-component of a 2nd generation smart fluorescent ligand. It has a single asparagine residue that is capable of displaying an N-linked glycan and a similar isoelectric point to ConA. Thus, binding between ConA and ovalbumin can potentially be monovalent and sugar specific. This work is the preliminary implementation of fluorescently-labeled ovalbumin in the ConA-based assay. We conjugate the red-emitting, long-lifetime azadioxatriangulenium (ADOTA+) dye to ovalbumin, as ADOTA have many advantageous properties to track the equilibrium binding of the assay. The ADOTA-labeled ovalbumin is paired with Alexa Fluor 647-labeled ConA to create a F{\"o}rster Resonance Energy Transfer (FRET) assay that is glucose dependent. The assay responds across the physiologically relevant glucose range (0-500 mg/dL) with increasing intensity from the ADOTA-ovalbumin, showing that the strategy may allow for the translation of the smart fluorescent ligand concept into a continuous glucose monitoring device.",
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Cummins, B, Simpson, J, Gryczynski, Z, Sørensen, TJ, Laursen, BW, Graham, D, Birch, D & Coté, G 2014, ConA-based glucose sensing using the long-lifetime azadioxatriangulenium fluorophore. in Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics., 89510A, Progress in Biomedical Optics and Imaging - Proceedings of SPIE, vol. 8951, SPIE, Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics, San Francisco, CA, United States, 3/02/14. https://doi.org/10.1117/12.2039824

ConA-based glucose sensing using the long-lifetime azadioxatriangulenium fluorophore. / Cummins, Brian; Simpson, Jonathan; Gryczynski, Zygmunt; Sørensen, Thomas Just; Laursen, Bo W.; Graham, Duncan; Birch, David; Coté, Gerard.

Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics. SPIE, 2014. 89510A (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 8951).

Research output: Chapter in Book/Report/Conference proceedingConference contributionResearchpeer-review

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T1 - ConA-based glucose sensing using the long-lifetime azadioxatriangulenium fluorophore

AU - Cummins, Brian

AU - Simpson, Jonathan

AU - Gryczynski, Zygmunt

AU - Sørensen, Thomas Just

AU - Laursen, Bo W.

AU - Graham, Duncan

AU - Birch, David

AU - Coté, Gerard

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N2 - Fluorescent glucose sensing technologies have been identified as possible alternatives to current continuous glucose monitoring approaches. We have recently introduced a new, smart fluorescent ligand to overcome the traditional problems of ConA-based glucose sensors. For this assay to be translated into a continuous glucose monitoring device where both components are free in solution, the molecular weight of the smart fluorescent ligand must be increased. We have identified ovalbumin as a naturally-occurring glycoprotein that could serve as the core-component of a 2nd generation smart fluorescent ligand. It has a single asparagine residue that is capable of displaying an N-linked glycan and a similar isoelectric point to ConA. Thus, binding between ConA and ovalbumin can potentially be monovalent and sugar specific. This work is the preliminary implementation of fluorescently-labeled ovalbumin in the ConA-based assay. We conjugate the red-emitting, long-lifetime azadioxatriangulenium (ADOTA+) dye to ovalbumin, as ADOTA have many advantageous properties to track the equilibrium binding of the assay. The ADOTA-labeled ovalbumin is paired with Alexa Fluor 647-labeled ConA to create a Förster Resonance Energy Transfer (FRET) assay that is glucose dependent. The assay responds across the physiologically relevant glucose range (0-500 mg/dL) with increasing intensity from the ADOTA-ovalbumin, showing that the strategy may allow for the translation of the smart fluorescent ligand concept into a continuous glucose monitoring device.

AB - Fluorescent glucose sensing technologies have been identified as possible alternatives to current continuous glucose monitoring approaches. We have recently introduced a new, smart fluorescent ligand to overcome the traditional problems of ConA-based glucose sensors. For this assay to be translated into a continuous glucose monitoring device where both components are free in solution, the molecular weight of the smart fluorescent ligand must be increased. We have identified ovalbumin as a naturally-occurring glycoprotein that could serve as the core-component of a 2nd generation smart fluorescent ligand. It has a single asparagine residue that is capable of displaying an N-linked glycan and a similar isoelectric point to ConA. Thus, binding between ConA and ovalbumin can potentially be monovalent and sugar specific. This work is the preliminary implementation of fluorescently-labeled ovalbumin in the ConA-based assay. We conjugate the red-emitting, long-lifetime azadioxatriangulenium (ADOTA+) dye to ovalbumin, as ADOTA have many advantageous properties to track the equilibrium binding of the assay. The ADOTA-labeled ovalbumin is paired with Alexa Fluor 647-labeled ConA to create a Förster Resonance Energy Transfer (FRET) assay that is glucose dependent. The assay responds across the physiologically relevant glucose range (0-500 mg/dL) with increasing intensity from the ADOTA-ovalbumin, showing that the strategy may allow for the translation of the smart fluorescent ligand concept into a continuous glucose monitoring device.

KW - Glucose sensing

KW - competitive binding

KW - energy transfer

KW - fluorescence

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PB - SPIE

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Cummins B, Simpson J, Gryczynski Z, Sørensen TJ, Laursen BW, Graham D et al. ConA-based glucose sensing using the long-lifetime azadioxatriangulenium fluorophore. In Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics. SPIE. 2014. 89510A. (Progress in Biomedical Optics and Imaging - Proceedings of SPIE). https://doi.org/10.1117/12.2039824