Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beads

Autores
Centi, Sonia; Messina, Germán Alejandro; Trombelli, Sara; Palchetti, Ilaria; Mascini, Marco
Año de publicación
2008
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Different assay formats based on the coupling of magnetic beads with electrochemical transduction were compared here for the detection of thrombin by using a thrombin specific aptamer. By using the thrombin-binding aptamer, a direct and an indirect competitive assay for thrombin have been developed by immobilising the aptamer or the protein, respectively. Moreover, another strategy was based on the direct measurement of the enzymatic product of thrombin captured by the immobilised aptamer. All the assays were developed by coupling the electrochemical transduction with the innovative and advantageous use of magnetic beads. The assays based on the immobilisation of the protein were not successful since no binding was recorded between thrombin and its aptamer. With the direct competitive assay, when the aptamer was immobilised onto the magnetic beads, a detection limit of 430 nM for thrombin was achieved. A lower detection limit for the protein (175 nM) was instead obtained by detecting the product of the enzymatic reaction catalysed by thrombin. All these assays were finally compared with a sandwich assay which reached a detection limit of 0.45 nM of thrombin demonstrating the best analytical performances. With this comparison the importance of a deep study on the different analytical approaches for thrombin detection to reach the performances of the best assay configuration has been demonstrated.
Fil: Centi, Sonia. Università degli Studi di Firenze; Italia
Fil: Messina, Germán Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Química de San Luis. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Instituto de Química de San Luis; Argentina. Università degli Studi di Firenze; Italia
Fil: Trombelli, Sara. Università degli Studi di Firenze; Italia
Fil: Palchetti, Ilaria. Università degli Studi di Firenze; Italia
Fil: Mascini, Marco. Università degli Studi di Firenze; Italia
Materia
APTAMER
THROMBIN
MAGNETIC BEADS
ELECTROCHEMICAL DETECTION
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/158404
Acceder
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beadsCenti, SoniaMessina, Germán AlejandroTrombelli, SaraPalchetti, IlariaMascini, MarcoAPTAMERTHROMBINMAGNETIC BEADSELECTROCHEMICAL DETECTIONhttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Different assay formats based on the coupling of magnetic beads with electrochemical transduction were compared here for the detection of thrombin by using a thrombin specific aptamer. By using the thrombin-binding aptamer, a direct and an indirect competitive assay for thrombin have been developed by immobilising the aptamer or the protein, respectively. Moreover, another strategy was based on the direct measurement of the enzymatic product of thrombin captured by the immobilised aptamer. All the assays were developed by coupling the electrochemical transduction with the innovative and advantageous use of magnetic beads. The assays based on the immobilisation of the protein were not successful since no binding was recorded between thrombin and its aptamer. With the direct competitive assay, when the aptamer was immobilised onto the magnetic beads, a detection limit of 430 nM for thrombin was achieved. A lower detection limit for the protein (175 nM) was instead obtained by detecting the product of the enzymatic reaction catalysed by thrombin. All these assays were finally compared with a sandwich assay which reached a detection limit of 0.45 nM of thrombin demonstrating the best analytical performances. With this comparison the importance of a deep study on the different analytical approaches for thrombin detection to reach the performances of the best assay configuration has been demonstrated.Fil: Centi, Sonia. Università degli Studi di Firenze; ItaliaFil: Messina, Germán Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Química de San Luis. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Instituto de Química de San Luis; Argentina. Università degli Studi di Firenze; ItaliaFil: Trombelli, Sara. Università degli Studi di Firenze; ItaliaFil: Palchetti, Ilaria. Università degli Studi di Firenze; ItaliaFil: Mascini, Marco. Università degli Studi di Firenze; ItaliaElsevier Advanced Technology2008-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/158404Centi, Sonia; Messina, Germán Alejandro; Trombelli, Sara; Palchetti, Ilaria; Mascini, Marco; Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beads; Elsevier Advanced Technology; Biosensors & Bioelectronics; 23; 11; 6-2008; 1602-16090956-5663CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S0956566308000390info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bios.2008.01.020info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:35:12Zoai:ri.conicet.gov.ar:11336/158404instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:35:12.442CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beads
title Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beads
spellingShingle Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beads
Centi, Sonia
APTAMER
THROMBIN
MAGNETIC BEADS
ELECTROCHEMICAL DETECTION
title_short Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beads
title_full Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beads
title_fullStr Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beads
title_full_unstemmed Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beads
title_sort Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beads
dc.creator.none.fl_str_mv Centi, Sonia
Messina, Germán Alejandro
Trombelli, Sara
Palchetti, Ilaria
Mascini, Marco
author Centi, Sonia
author_facet Centi, Sonia
Messina, Germán Alejandro
Trombelli, Sara
Palchetti, Ilaria
Mascini, Marco
author_role author
author2 Messina, Germán Alejandro
Trombelli, Sara
Palchetti, Ilaria
Mascini, Marco
author2_role author
author
author
author
dc.subject.none.fl_str_mv APTAMER
THROMBIN
MAGNETIC BEADS
ELECTROCHEMICAL DETECTION
topic APTAMER
THROMBIN
MAGNETIC BEADS
ELECTROCHEMICAL DETECTION
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Different assay formats based on the coupling of magnetic beads with electrochemical transduction were compared here for the detection of thrombin by using a thrombin specific aptamer. By using the thrombin-binding aptamer, a direct and an indirect competitive assay for thrombin have been developed by immobilising the aptamer or the protein, respectively. Moreover, another strategy was based on the direct measurement of the enzymatic product of thrombin captured by the immobilised aptamer. All the assays were developed by coupling the electrochemical transduction with the innovative and advantageous use of magnetic beads. The assays based on the immobilisation of the protein were not successful since no binding was recorded between thrombin and its aptamer. With the direct competitive assay, when the aptamer was immobilised onto the magnetic beads, a detection limit of 430 nM for thrombin was achieved. A lower detection limit for the protein (175 nM) was instead obtained by detecting the product of the enzymatic reaction catalysed by thrombin. All these assays were finally compared with a sandwich assay which reached a detection limit of 0.45 nM of thrombin demonstrating the best analytical performances. With this comparison the importance of a deep study on the different analytical approaches for thrombin detection to reach the performances of the best assay configuration has been demonstrated.
Fil: Centi, Sonia. Università degli Studi di Firenze; Italia
Fil: Messina, Germán Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Química de San Luis. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Instituto de Química de San Luis; Argentina. Università degli Studi di Firenze; Italia
Fil: Trombelli, Sara. Università degli Studi di Firenze; Italia
Fil: Palchetti, Ilaria. Università degli Studi di Firenze; Italia
Fil: Mascini, Marco. Università degli Studi di Firenze; Italia
description Different assay formats based on the coupling of magnetic beads with electrochemical transduction were compared here for the detection of thrombin by using a thrombin specific aptamer. By using the thrombin-binding aptamer, a direct and an indirect competitive assay for thrombin have been developed by immobilising the aptamer or the protein, respectively. Moreover, another strategy was based on the direct measurement of the enzymatic product of thrombin captured by the immobilised aptamer. All the assays were developed by coupling the electrochemical transduction with the innovative and advantageous use of magnetic beads. The assays based on the immobilisation of the protein were not successful since no binding was recorded between thrombin and its aptamer. With the direct competitive assay, when the aptamer was immobilised onto the magnetic beads, a detection limit of 430 nM for thrombin was achieved. A lower detection limit for the protein (175 nM) was instead obtained by detecting the product of the enzymatic reaction catalysed by thrombin. All these assays were finally compared with a sandwich assay which reached a detection limit of 0.45 nM of thrombin demonstrating the best analytical performances. With this comparison the importance of a deep study on the different analytical approaches for thrombin detection to reach the performances of the best assay configuration has been demonstrated.
publishDate 2008
dc.date.none.fl_str_mv 2008-06
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/158404
Centi, Sonia; Messina, Germán Alejandro; Trombelli, Sara; Palchetti, Ilaria; Mascini, Marco; Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beads; Elsevier Advanced Technology; Biosensors & Bioelectronics; 23; 11; 6-2008; 1602-1609
0956-5663
CONICET Digital
CONICET
url http://hdl.handle.net/11336/158404
identifier_str_mv Centi, Sonia; Messina, Germán Alejandro; Trombelli, Sara; Palchetti, Ilaria; Mascini, Marco; Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beads; Elsevier Advanced Technology; Biosensors & Bioelectronics; 23; 11; 6-2008; 1602-1609
0956-5663
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S0956566308000390
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bios.2008.01.020
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Advanced Technology
publisher.none.fl_str_mv Elsevier Advanced Technology
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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