Assessing the Covalent Attachment and Energy Transfer Capabilities of Upconverting Phosphors With Cofactor Containing Bioactive Enzymes

Upconverting phosphors (UCPs) convert multiple low energy photons into higher energy emission via the process of photon upconversion and offer an attractive alternative to organic fluorophores for use as luminescent probes. Examples of biosensors utilizing the apparent energy transfer of UCPs and na...

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Main Authors: Letitia Burgess, Hannah Wilson, Alex R. Jones, Sam Hay, Louise S. Natrajan
Format: Article
Language:English
Published: Frontiers Media S.A. 2020-12-01
Series:Frontiers in Chemistry
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fchem.2020.613334/full
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spelling doaj-2eb78352ef57490eb897a4d2bb539a562020-12-21T07:02:51ZengFrontiers Media S.A.Frontiers in Chemistry2296-26462020-12-01810.3389/fchem.2020.613334613334Assessing the Covalent Attachment and Energy Transfer Capabilities of Upconverting Phosphors With Cofactor Containing Bioactive EnzymesLetitia Burgess0Letitia Burgess1Hannah Wilson2Hannah Wilson3Hannah Wilson4Alex R. Jones5Alex R. Jones6Alex R. Jones7Sam Hay8Sam Hay9Louise S. Natrajan10Louise S. Natrajan11Department of Chemistry, School of Natural Sciences, The University of Manchester, Manchester, United KingdomManchester Institute of Biotechnology, The University of Manchester, Manchester, United KingdomDepartment of Chemistry, School of Natural Sciences, The University of Manchester, Manchester, United KingdomManchester Institute of Biotechnology, The University of Manchester, Manchester, United KingdomPhoton Science Institute, The University of Manchester, Manchester, United KingdomDepartment of Chemistry, School of Natural Sciences, The University of Manchester, Manchester, United KingdomManchester Institute of Biotechnology, The University of Manchester, Manchester, United KingdomPhoton Science Institute, The University of Manchester, Manchester, United KingdomDepartment of Chemistry, School of Natural Sciences, The University of Manchester, Manchester, United KingdomManchester Institute of Biotechnology, The University of Manchester, Manchester, United KingdomDepartment of Chemistry, School of Natural Sciences, The University of Manchester, Manchester, United KingdomPhoton Science Institute, The University of Manchester, Manchester, United KingdomUpconverting phosphors (UCPs) convert multiple low energy photons into higher energy emission via the process of photon upconversion and offer an attractive alternative to organic fluorophores for use as luminescent probes. Examples of biosensors utilizing the apparent energy transfer of UCPs and nanophosphors (UCNPs) with biomolecules have started to appear in the literature but very few exploit the covalent anchoring of the biomolecule to the surface of the UCP to improve the sensitivity of the systems. Here, we demonstrate a robust and versatile method for the covalent attachment of biomolecules to the surface of a variety of UCPs and UCNPs in which the UCPs were capped with functionalized silica in order to provide a surface to covalently conjugate biomolecules with surface-accessible cysteines. Variants of BM3Heme, cytochrome C, glucose oxidase, and glutathione reductase were then attached via maleimide-thiol coupling. BM3Heme, glucose oxidase, and glutathione reductase were shown to retain their activity when coupled to the UCPs potentially opening up opportunities for biosensing applications.https://www.frontiersin.org/articles/10.3389/fchem.2020.613334/fulllanthanidesupconversionluminescenceprotein conjugationenergy transferenzyme kinetics
collection DOAJ
language English
format Article
sources DOAJ
author Letitia Burgess
Letitia Burgess
Hannah Wilson
Hannah Wilson
Hannah Wilson
Alex R. Jones
Alex R. Jones
Alex R. Jones
Sam Hay
Sam Hay
Louise S. Natrajan
Louise S. Natrajan
spellingShingle Letitia Burgess
Letitia Burgess
Hannah Wilson
Hannah Wilson
Hannah Wilson
Alex R. Jones
Alex R. Jones
Alex R. Jones
Sam Hay
Sam Hay
Louise S. Natrajan
Louise S. Natrajan
Assessing the Covalent Attachment and Energy Transfer Capabilities of Upconverting Phosphors With Cofactor Containing Bioactive Enzymes
Frontiers in Chemistry
lanthanides
upconversion
luminescence
protein conjugation
energy transfer
enzyme kinetics
author_facet Letitia Burgess
Letitia Burgess
Hannah Wilson
Hannah Wilson
Hannah Wilson
Alex R. Jones
Alex R. Jones
Alex R. Jones
Sam Hay
Sam Hay
Louise S. Natrajan
Louise S. Natrajan
author_sort Letitia Burgess
title Assessing the Covalent Attachment and Energy Transfer Capabilities of Upconverting Phosphors With Cofactor Containing Bioactive Enzymes
title_short Assessing the Covalent Attachment and Energy Transfer Capabilities of Upconverting Phosphors With Cofactor Containing Bioactive Enzymes
title_full Assessing the Covalent Attachment and Energy Transfer Capabilities of Upconverting Phosphors With Cofactor Containing Bioactive Enzymes
title_fullStr Assessing the Covalent Attachment and Energy Transfer Capabilities of Upconverting Phosphors With Cofactor Containing Bioactive Enzymes
title_full_unstemmed Assessing the Covalent Attachment and Energy Transfer Capabilities of Upconverting Phosphors With Cofactor Containing Bioactive Enzymes
title_sort assessing the covalent attachment and energy transfer capabilities of upconverting phosphors with cofactor containing bioactive enzymes
publisher Frontiers Media S.A.
series Frontiers in Chemistry
issn 2296-2646
publishDate 2020-12-01
description Upconverting phosphors (UCPs) convert multiple low energy photons into higher energy emission via the process of photon upconversion and offer an attractive alternative to organic fluorophores for use as luminescent probes. Examples of biosensors utilizing the apparent energy transfer of UCPs and nanophosphors (UCNPs) with biomolecules have started to appear in the literature but very few exploit the covalent anchoring of the biomolecule to the surface of the UCP to improve the sensitivity of the systems. Here, we demonstrate a robust and versatile method for the covalent attachment of biomolecules to the surface of a variety of UCPs and UCNPs in which the UCPs were capped with functionalized silica in order to provide a surface to covalently conjugate biomolecules with surface-accessible cysteines. Variants of BM3Heme, cytochrome C, glucose oxidase, and glutathione reductase were then attached via maleimide-thiol coupling. BM3Heme, glucose oxidase, and glutathione reductase were shown to retain their activity when coupled to the UCPs potentially opening up opportunities for biosensing applications.
topic lanthanides
upconversion
luminescence
protein conjugation
energy transfer
enzyme kinetics
url https://www.frontiersin.org/articles/10.3389/fchem.2020.613334/full
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