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 contribution book

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. 

LanguageEnglish
Title of host publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Subtitle of host publicationOptical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics
EditorsGerard L. Coté
Place of PublicationBellingham
Volume8591
DOIs
Publication statusPublished - 28 Feb 2014
EventOptical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics - San Francisco, CA, United States
Duration: 3 Feb 20146 Feb 2014

Publication series

NameSPIE Proceedings
PublisherSPIE
Volume8951

Conference

ConferenceOptical 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

  • competitive binding
  • energy transfer
  • fluorescence
  • glucose sensing

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 G. L. Coté (Ed.), Progress in Biomedical Optics and Imaging - Proceedings of SPIE: Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics (Vol. 8591). [89510A] (SPIE Proceedings; Vol. 8951). Bellingham . 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. Progress in Biomedical Optics and Imaging - Proceedings of SPIE: Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics . editor / Gerard L. Coté. Vol. 8591 Bellingham , 2014. (SPIE Proceedings).
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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 GL Coté (ed.), Progress in Biomedical Optics and Imaging - Proceedings of SPIE: Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics . vol. 8591, 89510A, SPIE Proceedings, vol. 8951, Bellingham , 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.

Progress in Biomedical Optics and Imaging - Proceedings of SPIE: Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics . ed. / Gerard L. Coté. Vol. 8591 Bellingham , 2014. 89510A (SPIE Proceedings; Vol. 8951).

Research output: Chapter in Book/Report/Conference proceedingConference contribution book

TY - GEN

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

PY - 2014/2/28

Y1 - 2014/2/28

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 - competitive binding

KW - energy transfer

KW - fluorescence

KW - glucose sensing

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DO - 10.1117/12.2039824

M3 - Conference contribution book

SN - 9780819498649

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BT - Progress in Biomedical Optics and Imaging - Proceedings of SPIE

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ER -

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 Coté GL, editor, Progress in Biomedical Optics and Imaging - Proceedings of SPIE: Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics . Vol. 8591. Bellingham . 2014. 89510A. (SPIE Proceedings). https://doi.org/10.1117/12.2039824