Aqueous near infrared fluorescent composites based on apoferritin-encapsulated PbS quantum dots

Quantum dots offer a number of advantages over standard fluorescent dyes for monitoring biological systems including high luminescence, stability against photobleaching, and a wide range of fluorescence wavelengths from blue to infrared depending on the particle size. In this work, we investigated i...

Full description

Bibliographic Details
Main Author: Hennequin, Barbara
Format: Thesis (University of Nottingham only)
Language:English
Published: 2008
Subjects:
Online Access:https://eprints.nottingham.ac.uk/11071/
_version_ 1848791186942197760
author Hennequin, Barbara
author_facet Hennequin, Barbara
author_sort Hennequin, Barbara
building Nottingham Research Data Repository
collection Online Access
description Quantum dots offer a number of advantages over standard fluorescent dyes for monitoring biological systems including high luminescence, stability against photobleaching, and a wide range of fluorescence wavelengths from blue to infrared depending on the particle size. In this work, we investigated in using the protein cage apoferritin as a template for the synthesis of colloidal quantum dots. We obtained apoferritin after reductive dissolution of the ferritin iron core and showed that the protein structure was left intact during this process. We further studied the solubility of ferritin, apoferritin and cationized ferritin in organic and fluorinated solvents by hydrophobic ion pairing methodology in order to expand the possibility of using an apoferritin template for the synthesis of quantum dots in organic media. We then focused on the synthesis and fluorescence properties of PbS quantum dots in aqueous solution. PbS dots are thermally stable and emit in the range 1,100 to 1,300 nm depending on their size. We demonstrated the encapsulation of these PbS quantum dots within the cavity of the iron storage protein apoferritin using two routes: 1) the disassembly/reassembly of apoferritin subunits trapping previously synthesised PbS quantum dots; and 2) use of the channels present in the protein shell to allow the entrance of Pb2+ and S2- ions leading to formation of quantum dots in the apoferritin cavity. We show that PbS-apoferritin composites emit in the near infrared region which make them promising labels for biological applications. Furthermore, we demonstrated that PbS QDs can be excited via a bioluminescence resonance energy transfer (BRET) using luciferin from Luciola mingrelica which could be developed into a self-illuminating labelling system. Finally, in order to make PbS-apoferritin composites selectively attachable to biomolecules during labelling experiments, the apoferritin was modified by the incorporation of analogues of methionine introducing azido groups absent in the proteins. The azido groups can then be selectively modified in complex mixtures e.g. cell lysates using `bio-orthogonal' reactions such as the Cu(I) catalysed Staudinger ligation and Huisgen cycloaddition. This would allow highly selective addition of receptor targeting or cellular permeation of peptides to the outer surface of the apoferritin shell.
first_indexed 2025-11-14T18:24:31Z
format Thesis (University of Nottingham only)
id nottingham-11071
institution University of Nottingham Malaysia Campus
institution_category Local University
language English
last_indexed 2025-11-14T18:24:31Z
publishDate 2008
recordtype eprints
repository_type Digital Repository
spelling nottingham-110712025-02-28T11:11:05Z https://eprints.nottingham.ac.uk/11071/ Aqueous near infrared fluorescent composites based on apoferritin-encapsulated PbS quantum dots Hennequin, Barbara Quantum dots offer a number of advantages over standard fluorescent dyes for monitoring biological systems including high luminescence, stability against photobleaching, and a wide range of fluorescence wavelengths from blue to infrared depending on the particle size. In this work, we investigated in using the protein cage apoferritin as a template for the synthesis of colloidal quantum dots. We obtained apoferritin after reductive dissolution of the ferritin iron core and showed that the protein structure was left intact during this process. We further studied the solubility of ferritin, apoferritin and cationized ferritin in organic and fluorinated solvents by hydrophobic ion pairing methodology in order to expand the possibility of using an apoferritin template for the synthesis of quantum dots in organic media. We then focused on the synthesis and fluorescence properties of PbS quantum dots in aqueous solution. PbS dots are thermally stable and emit in the range 1,100 to 1,300 nm depending on their size. We demonstrated the encapsulation of these PbS quantum dots within the cavity of the iron storage protein apoferritin using two routes: 1) the disassembly/reassembly of apoferritin subunits trapping previously synthesised PbS quantum dots; and 2) use of the channels present in the protein shell to allow the entrance of Pb2+ and S2- ions leading to formation of quantum dots in the apoferritin cavity. We show that PbS-apoferritin composites emit in the near infrared region which make them promising labels for biological applications. Furthermore, we demonstrated that PbS QDs can be excited via a bioluminescence resonance energy transfer (BRET) using luciferin from Luciola mingrelica which could be developed into a self-illuminating labelling system. Finally, in order to make PbS-apoferritin composites selectively attachable to biomolecules during labelling experiments, the apoferritin was modified by the incorporation of analogues of methionine introducing azido groups absent in the proteins. The azido groups can then be selectively modified in complex mixtures e.g. cell lysates using `bio-orthogonal' reactions such as the Cu(I) catalysed Staudinger ligation and Huisgen cycloaddition. This would allow highly selective addition of receptor targeting or cellular permeation of peptides to the outer surface of the apoferritin shell. 2008-12-11 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en arr https://eprints.nottingham.ac.uk/11071/1/Thesis_B._Hennequin.pdf Hennequin, Barbara (2008) Aqueous near infrared fluorescent composites based on apoferritin-encapsulated PbS quantum dots. PhD thesis, University of Nottingham. Quantum dots PbS apoferritin photoluminescence colloidal nanocrystals template near infrared BRET un-natural amino acid.
spellingShingle Quantum dots
PbS
apoferritin
photoluminescence
colloidal nanocrystals
template
near infrared
BRET
un-natural amino acid.
Hennequin, Barbara
Aqueous near infrared fluorescent composites based on apoferritin-encapsulated PbS quantum dots
title Aqueous near infrared fluorescent composites based on apoferritin-encapsulated PbS quantum dots
title_full Aqueous near infrared fluorescent composites based on apoferritin-encapsulated PbS quantum dots
title_fullStr Aqueous near infrared fluorescent composites based on apoferritin-encapsulated PbS quantum dots
title_full_unstemmed Aqueous near infrared fluorescent composites based on apoferritin-encapsulated PbS quantum dots
title_short Aqueous near infrared fluorescent composites based on apoferritin-encapsulated PbS quantum dots
title_sort aqueous near infrared fluorescent composites based on apoferritin-encapsulated pbs quantum dots
topic Quantum dots
PbS
apoferritin
photoluminescence
colloidal nanocrystals
template
near infrared
BRET
un-natural amino acid.
url https://eprints.nottingham.ac.uk/11071/