Solution-phase vs surface-phase aptamer-protein affinity from a label-free kinetic biosensor.

Aptamers are selected DNA ligands that target biomolecules such as proteins. In recent years, they are showing an increasing interest as potential therapeutic agents or recognition elements in biosensor applications. In both cases, the need for characterizing the mating between the target and the ap...

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Autores principales:	Camille Daniel, Yoann Roupioz, Didier Gasparutto, Thierry Livache, Arnaud Buhot
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/576a4e0622734ecf8d724d58ccded4c0
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spelling	oai:doaj.org-article:576a4e0622734ecf8d724d58ccded4c02021-11-18T08:54:51ZSolution-phase vs surface-phase aptamer-protein affinity from a label-free kinetic biosensor.1932-620310.1371/journal.pone.0075419https://doaj.org/article/576a4e0622734ecf8d724d58ccded4c02013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24069412/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Aptamers are selected DNA ligands that target biomolecules such as proteins. In recent years, they are showing an increasing interest as potential therapeutic agents or recognition elements in biosensor applications. In both cases, the need for characterizing the mating between the target and the aptamer either in solution or immobilized on a surface, is pressing. In this context, we have developed a kinetic biosensor made of micro-arrayed anti-thrombin aptamers to assess the kinetic parameters of this interaction. The binding of label-free thrombin on the biosensor was monitored in real-time by Surface Plasmon Resonance imaging. Remarkable performances were obtained for the quantification of thrombin without amplification (sub-nanomolar limit of detection and linear range of quantification to two orders of magnitude). The independent determinations of both the solution- and surface-phase affinities, respectively KD(Sol) and KD(Surf), revealed distinct values illustrating the importance of probes, targets or surface interactions in biosensors. Interestingly, KD(Surf) values depend on the aptamer grafting density and linearly extrapolate towards KD(Sol) for highly diluted probes. This suggests a lesser impact of the surface compared to the probe or target cooperativity interactions since the latter decrease with a reduced grafting density.Camille DanielYoann RoupiozDidier GasparuttoThierry LivacheArnaud BuhotPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 9, p e75419 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Camille Daniel Yoann Roupioz Didier Gasparutto Thierry Livache Arnaud Buhot Solution-phase vs surface-phase aptamer-protein affinity from a label-free kinetic biosensor.
description	Aptamers are selected DNA ligands that target biomolecules such as proteins. In recent years, they are showing an increasing interest as potential therapeutic agents or recognition elements in biosensor applications. In both cases, the need for characterizing the mating between the target and the aptamer either in solution or immobilized on a surface, is pressing. In this context, we have developed a kinetic biosensor made of micro-arrayed anti-thrombin aptamers to assess the kinetic parameters of this interaction. The binding of label-free thrombin on the biosensor was monitored in real-time by Surface Plasmon Resonance imaging. Remarkable performances were obtained for the quantification of thrombin without amplification (sub-nanomolar limit of detection and linear range of quantification to two orders of magnitude). The independent determinations of both the solution- and surface-phase affinities, respectively KD(Sol) and KD(Surf), revealed distinct values illustrating the importance of probes, targets or surface interactions in biosensors. Interestingly, KD(Surf) values depend on the aptamer grafting density and linearly extrapolate towards KD(Sol) for highly diluted probes. This suggests a lesser impact of the surface compared to the probe or target cooperativity interactions since the latter decrease with a reduced grafting density.
format	article
author	Camille Daniel Yoann Roupioz Didier Gasparutto Thierry Livache Arnaud Buhot
author_facet	Camille Daniel Yoann Roupioz Didier Gasparutto Thierry Livache Arnaud Buhot
author_sort	Camille Daniel
title	Solution-phase vs surface-phase aptamer-protein affinity from a label-free kinetic biosensor.
title_short	Solution-phase vs surface-phase aptamer-protein affinity from a label-free kinetic biosensor.
title_full	Solution-phase vs surface-phase aptamer-protein affinity from a label-free kinetic biosensor.
title_fullStr	Solution-phase vs surface-phase aptamer-protein affinity from a label-free kinetic biosensor.
title_full_unstemmed	Solution-phase vs surface-phase aptamer-protein affinity from a label-free kinetic biosensor.
title_sort	solution-phase vs surface-phase aptamer-protein affinity from a label-free kinetic biosensor.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/576a4e0622734ecf8d724d58ccded4c0
work_keys_str_mv	AT camilledaniel solutionphasevssurfacephaseaptamerproteinaffinityfromalabelfreekineticbiosensor AT yoannroupioz solutionphasevssurfacephaseaptamerproteinaffinityfromalabelfreekineticbiosensor AT didiergasparutto solutionphasevssurfacephaseaptamerproteinaffinityfromalabelfreekineticbiosensor AT thierrylivache solutionphasevssurfacephaseaptamerproteinaffinityfromalabelfreekineticbiosensor AT arnaudbuhot solutionphasevssurfacephaseaptamerproteinaffinityfromalabelfreekineticbiosensor
_version_	1718421141632057344

Solution-phase vs surface-phase aptamer-protein affinity from a label-free kinetic biosensor.

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