Signal amplification for thrombin impedimetric aptasensor

In this work, we report a highly specific amplification strategy demonstrated for the ultrasensitive biosensing of thrombin with the use of gold-streptavidin nanoparticles (strep-AuNPs) and silver reduction enhancement. The biotinylated aptamer of thrombin was immobilized onto an avidin-graphite epo...

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Autores: Ocaña Tejada, Cristina, Valle, Manel del|||0000-0002-1032-8611
Tipo de recurso: artículo
Fecha de publicación:2014
País:España
Institución:Universitat Autònoma de Barcelona
Repositorio:Dipòsit Digital de Documents de la UAB
Idioma:inglés
OAI Identifier:oai:ddd.uab.cat:154803
Acceso en línea:https://ddd.uab.cat/record/154803
https://dx.doi.org/urn:doi:10.1016/j.bios.2013.10.068
Access Level:acceso abierto
Palabra clave:Impedance
Nanoparticles
Aptamer
Biosensor
Thrombin
Quantum dots
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spelling Signal amplification for thrombin impedimetric aptasensorsandwich protocol and use of gold-streptavidin nanoparticlesOcaña Tejada, CristinaValle, Manel del|||0000-0002-1032-8611ImpedanceNanoparticlesAptamerBiosensorThrombinQuantum dotsIn this work, we report a highly specific amplification strategy demonstrated for the ultrasensitive biosensing of thrombin with the use of gold-streptavidin nanoparticles (strep-AuNPs) and silver reduction enhancement. The biotinylated aptamer of thrombin was immobilized onto an avidin-graphite epoxy composite (AvGEC) electrode surface by affinity interaction between biotin and avidin; electrochemical impedance measurements were performed in a solution containing the redox marker ferrocyanide/ferricyanide. The change in interfacial charge transfer resistance (Rct) experimented by the redox marker, was recorded to confirm aptamer complex formation with target protein, thrombin (Thr), in a label-free first stage. A biotinylated second thrombin aptamer, with complementary recognition properties was then used in a sandwich approach. The addition of strep-AuNPs and silver enhancement treatment led to a further increment of Rct thus obtaining significant signal amplification. The AptThrBio1-Thr-AptThrBio2 sandwich formation was inspected by confocal microcopy after incubation with streptavidin quantum dots. In order to visualize the presence of gold nanoparticles, the same silver enhancement treatment was applied to electrodes already modified with the nanoparticle-sandwich conjugate, allowing direct observation by scanning electron microscopy (SEM). Results showed high sensitivity and selectivity for thrombin detection, with an improvement from ca. 4.7 pM in a simple assay to 0.3 pM in the amplified reported scheme. 22014-01-0120142014-01-01Articlehttp://purl.org/coar/resource_type/c_6501SMURhttp://purl.org/coar/version/c_71e4c1898caa6e32info:eu-repo/semantics/articleapplication/pdfhttps://ddd.uab.cat/record/154803https://dx.doi.org/urn:doi:10.1016/j.bios.2013.10.068reponame:Dipòsit Digital de Documents de la UABinstname:Universitat Autònoma de BarcelonaInglésengMinisterio de Ciencia e Innovación https://doi.org/10.13039/501100004837 CTQ2010-17099open accesshttp://purl.org/coar/access_right/c_abf2Aquest material està protegit per drets d'autor i/o drets afins. Podeu utilitzar aquest material en funció del que permet la legislació de drets d'autor i drets afins d'aplicació al vostre cas. Per a d'altres usos heu d'obtenir permís del(s) titular(s) de drets.https://rightsstatements.org/vocab/InC/1.0/info:eu-repo/semantics/openAccessoai:ddd.uab.cat:1548032026-06-06T12:50:31Z
dc.title.none.fl_str_mv Signal amplification for thrombin impedimetric aptasensor
sandwich protocol and use of gold-streptavidin nanoparticles
title Signal amplification for thrombin impedimetric aptasensor
spellingShingle Signal amplification for thrombin impedimetric aptasensor
Ocaña Tejada, Cristina
Impedance
Nanoparticles
Aptamer
Biosensor
Thrombin
Quantum dots
title_short Signal amplification for thrombin impedimetric aptasensor
title_full Signal amplification for thrombin impedimetric aptasensor
title_fullStr Signal amplification for thrombin impedimetric aptasensor
title_full_unstemmed Signal amplification for thrombin impedimetric aptasensor
title_sort Signal amplification for thrombin impedimetric aptasensor
dc.creator.none.fl_str_mv Ocaña Tejada, Cristina
Valle, Manel del|||0000-0002-1032-8611
author Ocaña Tejada, Cristina
author_facet Ocaña Tejada, Cristina
Valle, Manel del|||0000-0002-1032-8611
author_role author
author2 Valle, Manel del|||0000-0002-1032-8611
author2_role author
dc.subject.none.fl_str_mv Impedance
Nanoparticles
Aptamer
Biosensor
Thrombin
Quantum dots
topic Impedance
Nanoparticles
Aptamer
Biosensor
Thrombin
Quantum dots
description In this work, we report a highly specific amplification strategy demonstrated for the ultrasensitive biosensing of thrombin with the use of gold-streptavidin nanoparticles (strep-AuNPs) and silver reduction enhancement. The biotinylated aptamer of thrombin was immobilized onto an avidin-graphite epoxy composite (AvGEC) electrode surface by affinity interaction between biotin and avidin; electrochemical impedance measurements were performed in a solution containing the redox marker ferrocyanide/ferricyanide. The change in interfacial charge transfer resistance (Rct) experimented by the redox marker, was recorded to confirm aptamer complex formation with target protein, thrombin (Thr), in a label-free first stage. A biotinylated second thrombin aptamer, with complementary recognition properties was then used in a sandwich approach. The addition of strep-AuNPs and silver enhancement treatment led to a further increment of Rct thus obtaining significant signal amplification. The AptThrBio1-Thr-AptThrBio2 sandwich formation was inspected by confocal microcopy after incubation with streptavidin quantum dots. In order to visualize the presence of gold nanoparticles, the same silver enhancement treatment was applied to electrodes already modified with the nanoparticle-sandwich conjugate, allowing direct observation by scanning electron microscopy (SEM). Results showed high sensitivity and selectivity for thrombin detection, with an improvement from ca. 4.7 pM in a simple assay to 0.3 pM in the amplified reported scheme.
publishDate 2014
dc.date.none.fl_str_mv 2
2014-01-01
2014
2014-01-01
dc.type.none.fl_str_mv Article
http://purl.org/coar/resource_type/c_6501
SMUR
http://purl.org/coar/version/c_71e4c1898caa6e32
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://ddd.uab.cat/record/154803
https://dx.doi.org/urn:doi:10.1016/j.bios.2013.10.068
url https://ddd.uab.cat/record/154803
https://dx.doi.org/urn:doi:10.1016/j.bios.2013.10.068
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Ministerio de Ciencia e Innovación https://doi.org/10.13039/501100004837 CTQ2010-17099
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
https://rightsstatements.org/vocab/InC/1.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
https://rightsstatements.org/vocab/InC/1.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Dipòsit Digital de Documents de la UAB
instname:Universitat Autònoma de Barcelona
instname_str Universitat Autònoma de Barcelona
reponame_str Dipòsit Digital de Documents de la UAB
collection Dipòsit Digital de Documents de la UAB
repository.name.fl_str_mv
repository.mail.fl_str_mv
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