Modular fluorescent nanoparticle DNA probes for detection of peptides and proteins

Abstract Fluorescently labeled antibody and aptamer probes are used in biological studies to characterize binding interactions, measure concentrations of analytes, and sort cells. Fluorescent nanoparticle labels offer an excellent alternative to standard fluorescent labeling strategies due to their...

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Autores principales:	Cassandra M. Stawicki, Torri E. Rinker, Markus Burns, Sonal S. Tonapi, Rachel P. Galimidi, Deepthi Anumala, Julia K. Robinson, Joshua S. Klein, Parag Mallick
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1d1df3ba9f9349db96cddc7b9dd3b6da
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spelling	oai:doaj.org-article:1d1df3ba9f9349db96cddc7b9dd3b6da2021-12-02T18:37:10ZModular fluorescent nanoparticle DNA probes for detection of peptides and proteins10.1038/s41598-021-99084-42045-2322https://doaj.org/article/1d1df3ba9f9349db96cddc7b9dd3b6da2021-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-99084-4https://doaj.org/toc/2045-2322Abstract Fluorescently labeled antibody and aptamer probes are used in biological studies to characterize binding interactions, measure concentrations of analytes, and sort cells. Fluorescent nanoparticle labels offer an excellent alternative to standard fluorescent labeling strategies due to their enhanced brightness, stability and multivalency; however, challenges in functionalization and characterization have impeded their use. This work introduces a straightforward approach for preparation of fluorescent nanoparticle probes using commercially available reagents and common laboratory equipment. Fluorescent polystyrene nanoparticles, Thermo Fisher Scientific FluoSpheres, were used in these proof-of-principle studies. Particle passivation was achieved by covalent attachment of amine-PEG-azide to carboxylated particles, neutralizing the surface charge from − 43 to − 15 mV. A conjugation-annealing handle and DNA aptamer probe were attached to the azide-PEG nanoparticle surface either through reaction of pre-annealed handle and probe or through a stepwise reaction of the nanoparticles with the handle followed by aptamer annealing. Nanoparticles functionalized with DNA aptamers targeting histidine tags and VEGF protein had high affinity (EC50s ranging from 3 to 12 nM) and specificity, and were more stable than conventional labels. This protocol for preparation of nanoparticle probes relies solely on commercially available reagents and common equipment, breaking down the barriers to use nanoparticles in biological experiments.Cassandra M. StawickiTorri E. RinkerMarkus BurnsSonal S. TonapiRachel P. GalimidiDeepthi AnumalaJulia K. RobinsonJoshua S. KleinParag MallickNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Cassandra M. Stawicki Torri E. Rinker Markus Burns Sonal S. Tonapi Rachel P. Galimidi Deepthi Anumala Julia K. Robinson Joshua S. Klein Parag Mallick Modular fluorescent nanoparticle DNA probes for detection of peptides and proteins
description	Abstract Fluorescently labeled antibody and aptamer probes are used in biological studies to characterize binding interactions, measure concentrations of analytes, and sort cells. Fluorescent nanoparticle labels offer an excellent alternative to standard fluorescent labeling strategies due to their enhanced brightness, stability and multivalency; however, challenges in functionalization and characterization have impeded their use. This work introduces a straightforward approach for preparation of fluorescent nanoparticle probes using commercially available reagents and common laboratory equipment. Fluorescent polystyrene nanoparticles, Thermo Fisher Scientific FluoSpheres, were used in these proof-of-principle studies. Particle passivation was achieved by covalent attachment of amine-PEG-azide to carboxylated particles, neutralizing the surface charge from − 43 to − 15 mV. A conjugation-annealing handle and DNA aptamer probe were attached to the azide-PEG nanoparticle surface either through reaction of pre-annealed handle and probe or through a stepwise reaction of the nanoparticles with the handle followed by aptamer annealing. Nanoparticles functionalized with DNA aptamers targeting histidine tags and VEGF protein had high affinity (EC50s ranging from 3 to 12 nM) and specificity, and were more stable than conventional labels. This protocol for preparation of nanoparticle probes relies solely on commercially available reagents and common equipment, breaking down the barriers to use nanoparticles in biological experiments.
format	article
author	Cassandra M. Stawicki Torri E. Rinker Markus Burns Sonal S. Tonapi Rachel P. Galimidi Deepthi Anumala Julia K. Robinson Joshua S. Klein Parag Mallick
author_facet	Cassandra M. Stawicki Torri E. Rinker Markus Burns Sonal S. Tonapi Rachel P. Galimidi Deepthi Anumala Julia K. Robinson Joshua S. Klein Parag Mallick
author_sort	Cassandra M. Stawicki
title	Modular fluorescent nanoparticle DNA probes for detection of peptides and proteins
title_short	Modular fluorescent nanoparticle DNA probes for detection of peptides and proteins
title_full	Modular fluorescent nanoparticle DNA probes for detection of peptides and proteins
title_fullStr	Modular fluorescent nanoparticle DNA probes for detection of peptides and proteins
title_full_unstemmed	Modular fluorescent nanoparticle DNA probes for detection of peptides and proteins
title_sort	modular fluorescent nanoparticle dna probes for detection of peptides and proteins
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/1d1df3ba9f9349db96cddc7b9dd3b6da
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Modular fluorescent nanoparticle DNA probes for detection of peptides and proteins

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