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Nano Research 2019, 12(8): 1862-1870 https://doi.org/10.1007/s12274-019-2449-x
Research Article | Issue | Published: 18 June 2019

Amine-rich carbon nitride mnanoparticles: Synthesis, covalent functionalization with proteins and application in a fluorescence quenching assay

Show Author's Information Gabriele Capilli†,  Simone Cavalera,  Laura Anfossi( ),  Cristina Giovannoli,  Marco Minella( ),  Claudio Baggiani,  Claudio Minero
Department of Chemistry and NIS Center of Excellence,University of Torino,10125,Torino, Italy;

†Present address: Mining & Materials Engineering, McGill University, Montreal, Quebec H3A 0C5, Canada

Keywords:
g-C3N4, fluorescent probe, carbon nitride nanoparticles, quenching assay, fluorescent materials, Forster resonant energy transfer (FRET)
Topics:
BiosynthesisCarbon nitrideFluorescence quenchingFluorescence spectroscopyFluorophoresHigh resolution transmission electron microscopyIrradiationMetabolismNanoparticlesNitridesOptical propertiesProbesProteinsQuantum yield
Cite this article:
Capilli G, Cavalera S, Anfossi L, et al. Amine-rich carbon nitride mnanoparticles: Synthesis, covalent functionalization with proteins and application in a fluorescence quenching assay. Nano Research, 2019, 12(8): 1862-1870. https://doi.org/10.1007/s12274-019-2449-x
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Abstract Full text Electronic supplementary material About this article

Carbon nitride nanoparticles (CNNPs) have been employed as fluorescent sensing tools owing to their unique features, e.g. low cost production, high stability in water and high photoluminescence quantum yield. Here, an easy and versatile synthetic approach was exploited to design fluorescent nanoparticles with surface functionalities suitable for covalent binding to bioligands. High hydrophilic, brightly fluorescent CNNPs, rich of superficial amines, were obtained from the thermal condensation of urea and lysine (CNNPLys) and by tuning the precursor ratio and the heating time. Structure and size of the functionalized nanoparticles were characterized through infrared (IR) spectroscopy, transmission electron microscopy (TEM) and dynamic light scattering (DLS). Their optical properties were studied by ultravioletɃvisible (UVɃVis) and fluorescence spectroscopy. The superficial primary amino groups, furnished by the lysine co-precursor, enabled for covalently linking CNNPLys to model proteins. The CNNPLys-protein conjugates excited under UV irradiation emit in the 400Ƀ450 nm visible range (quantum yield 24%). The applicability of CNNPLys as novel fluorescent probes was demonstrated by a fluorescence quenching assay, in which gold nanoparticles (GNPs) were attached to Staphylococcal protein A and employed to quench the CNNPLys fluorescence by Forster resonant energy transfer (FRET). The quenching occurred upon formation of the specific binding between the GNP-linked protein A and CNNPLys-tagged immunoglobulins, while the inhibition of the binding resulted in the recovery of CNNPLys luminescence. The synthetic strategy, based on combining a pconjugated polymerq-forming unit (urea) and a co-precursor able to provide the desired functional group (lysine), allows designing innovative materials for the development of new generation fluorescence biosensors in which easily functionalized fluorophores are needed.


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Electronic supplementary material
About this article

Amine-rich carbon nitride mnanoparticles: Synthesis, covalent functionalization with proteins and application in a fluorescence quenching assay

Show Author's information Gabriele Capilli†, Simone Cavalera, Laura Anfossi( ), Cristina Giovannoli, Marco Minella( ), Claudio Baggiani, Claudio Minero
Department of Chemistry and NIS Center of Excellence,University of Torino,10125,Torino, Italy;

†Present address: Mining & Materials Engineering, McGill University, Montreal, Quebec H3A 0C5, Canada

Abstract

Carbon nitride nanoparticles (CNNPs) have been employed as fluorescent sensing tools owing to their unique features, e.g. low cost production, high stability in water and high photoluminescence quantum yield. Here, an easy and versatile synthetic approach was exploited to design fluorescent nanoparticles with surface functionalities suitable for covalent binding to bioligands. High hydrophilic, brightly fluorescent CNNPs, rich of superficial amines, were obtained from the thermal condensation of urea and lysine (CNNPLys) and by tuning the precursor ratio and the heating time. Structure and size of the functionalized nanoparticles were characterized through infrared (IR) spectroscopy, transmission electron microscopy (TEM) and dynamic light scattering (DLS). Their optical properties were studied by ultravioletɃvisible (UVɃVis) and fluorescence spectroscopy. The superficial primary amino groups, furnished by the lysine co-precursor, enabled for covalently linking CNNPLys to model proteins. The CNNPLys-protein conjugates excited under UV irradiation emit in the 400Ƀ450 nm visible range (quantum yield 24%). The applicability of CNNPLys as novel fluorescent probes was demonstrated by a fluorescence quenching assay, in which gold nanoparticles (GNPs) were attached to Staphylococcal protein A and employed to quench the CNNPLys fluorescence by Forster resonant energy transfer (FRET). The quenching occurred upon formation of the specific binding between the GNP-linked protein A and CNNPLys-tagged immunoglobulins, while the inhibition of the binding resulted in the recovery of CNNPLys luminescence. The synthetic strategy, based on combining a pconjugated polymerq-forming unit (urea) and a co-precursor able to provide the desired functional group (lysine), allows designing innovative materials for the development of new generation fluorescence biosensors in which easily functionalized fluorophores are needed.

Keywords: g-C3N4, fluorescent probe, carbon nitride nanoparticles, quenching assay, fluorescent materials, Forster resonant energy transfer (FRET)

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Publication history
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Acknowledgements

Publication history

Received: 20 December 2018
Revised: 28 May 2019
Accepted: 30 May 2019
Published: 18 June 2019
Issue date: August 2019

Copyright

© Tsinghua University Press and Springer-Verlag GmbH Germany, part of Springer Nature 2019

Acknowledgements

Acknowledgements

The financial support from project Ricerca Locale-Torino University is gratefully acknowledged.

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