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Cadmium-Free Quantum Dots as Fluorescent Labels for Exosomes

Quantum dots are attractive alternatives to organic fluorophores for the purposes of fluorescent labeling and the detection of biomarkers. They can also be made to specifically target a protein of interest by conjugating biomolecules, such as antibodies. However, the majority of the fluorescent labe...

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Autores principales: Dobhal, Garima, Ayupova, Deanna, Laufersky, Geoffry, Ayed, Zeineb, Nann, Thomas, Goreham, Renee V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6210340/
https://www.ncbi.nlm.nih.gov/pubmed/30279349
http://dx.doi.org/10.3390/s18103308
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author Dobhal, Garima
Ayupova, Deanna
Laufersky, Geoffry
Ayed, Zeineb
Nann, Thomas
Goreham, Renee V.
author_facet Dobhal, Garima
Ayupova, Deanna
Laufersky, Geoffry
Ayed, Zeineb
Nann, Thomas
Goreham, Renee V.
author_sort Dobhal, Garima
collection PubMed
description Quantum dots are attractive alternatives to organic fluorophores for the purposes of fluorescent labeling and the detection of biomarkers. They can also be made to specifically target a protein of interest by conjugating biomolecules, such as antibodies. However, the majority of the fluorescent labeling using quantum dots is done using toxic materials such as cadmium or lead due to the well-established synthetic processes for these quantum dots. Here, we demonstrate the use of indium phosphide quantum dots with a zinc sulfide shell for the purposes of labeling and the detection of exosomes derived from the THP-1 cell line (monocyte cell line). Exosomes are nano-sized vesicles that have the potential to be used as biomarkers due to their involvement in complex cell processes. However, the lack of standardized methodology around the detection and analysis of exosomes has made it difficult to detect these membrane-containing vesicles. We targeted a protein that is known to exist on the surface of the exosomes (CD63) using a CD63 antibody. The antibody was conjugated to the quantum dots that were first made water-soluble using a ligand-exchange method. The conjugation was done using carbodiimide coupling, and was confirmed using a range of different methods such as dynamic light scattering, surface plasmon resonance, fluorescent microscopy, and Fourier transform infrared spectroscopy. The conjugation of the quantum dot antibody to the exosomes was further confirmed using similar methods. This demonstrates the potential for the use of a non-toxic conjugate to target nano-sized biomarkers that could be further used for the detection of different diseases.
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spelling pubmed-62103402018-11-02 Cadmium-Free Quantum Dots as Fluorescent Labels for Exosomes Dobhal, Garima Ayupova, Deanna Laufersky, Geoffry Ayed, Zeineb Nann, Thomas Goreham, Renee V. Sensors (Basel) Article Quantum dots are attractive alternatives to organic fluorophores for the purposes of fluorescent labeling and the detection of biomarkers. They can also be made to specifically target a protein of interest by conjugating biomolecules, such as antibodies. However, the majority of the fluorescent labeling using quantum dots is done using toxic materials such as cadmium or lead due to the well-established synthetic processes for these quantum dots. Here, we demonstrate the use of indium phosphide quantum dots with a zinc sulfide shell for the purposes of labeling and the detection of exosomes derived from the THP-1 cell line (monocyte cell line). Exosomes are nano-sized vesicles that have the potential to be used as biomarkers due to their involvement in complex cell processes. However, the lack of standardized methodology around the detection and analysis of exosomes has made it difficult to detect these membrane-containing vesicles. We targeted a protein that is known to exist on the surface of the exosomes (CD63) using a CD63 antibody. The antibody was conjugated to the quantum dots that were first made water-soluble using a ligand-exchange method. The conjugation was done using carbodiimide coupling, and was confirmed using a range of different methods such as dynamic light scattering, surface plasmon resonance, fluorescent microscopy, and Fourier transform infrared spectroscopy. The conjugation of the quantum dot antibody to the exosomes was further confirmed using similar methods. This demonstrates the potential for the use of a non-toxic conjugate to target nano-sized biomarkers that could be further used for the detection of different diseases. MDPI 2018-10-02 /pmc/articles/PMC6210340/ /pubmed/30279349 http://dx.doi.org/10.3390/s18103308 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Dobhal, Garima
Ayupova, Deanna
Laufersky, Geoffry
Ayed, Zeineb
Nann, Thomas
Goreham, Renee V.
Cadmium-Free Quantum Dots as Fluorescent Labels for Exosomes
title Cadmium-Free Quantum Dots as Fluorescent Labels for Exosomes
title_full Cadmium-Free Quantum Dots as Fluorescent Labels for Exosomes
title_fullStr Cadmium-Free Quantum Dots as Fluorescent Labels for Exosomes
title_full_unstemmed Cadmium-Free Quantum Dots as Fluorescent Labels for Exosomes
title_short Cadmium-Free Quantum Dots as Fluorescent Labels for Exosomes
title_sort cadmium-free quantum dots as fluorescent labels for exosomes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6210340/
https://www.ncbi.nlm.nih.gov/pubmed/30279349
http://dx.doi.org/10.3390/s18103308
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