Integrated amperometric affinity biosensors using Co2+-tetradentane nitrilotriacetic acid modified disposable carbon electrodes. Application to the determination of beta-lactam antibiotics
A novel strategy for the construction of disposable amperometric affinity biosensors is described in this work. The approach uses a recombinant bacterial penicillin binding protein (PBP) tagged by an N-terminal hexahistidine tail which was immobilized onto Co2+−tetradentate nitrilotriacetic acid (NTA)-modified screen-printed carbon electrodes (SPCEs). The biosensor was employed for the specific detection and quantification of β-lactam antibiotics residues in milk, which was accomplished by means of a direct competitive assay using a tracer with horseradish peroxidase (HRP) for the enzymatic labeling. The amperometric response measured at −0.20 V versus the Ag pseudoreference electrode of the SPCE upon the addition of H2O2 in the presence of hydroquinone (HQ) as redox mediator was used as the transduction signal. The developed affinity sensor allowed limits of detection to be obtained in the low part-per-billion level for the antibiotics tested in untreated milk samples. Moreover, the biosensor exhibited a good selectivity against other antibiotics residues frequently detected in milk and dairy products. The analysis time was of approximately 30 min.
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American Chemical Society
2013
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Online Access: | http://hdl.handle.net/10261/89711 http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100003176 http://dx.doi.org/10.13039/100012818 |
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dig-ictan-es-10261-897112021-06-30T07:49:21Z Integrated amperometric affinity biosensors using Co2+-tetradentane nitrilotriacetic acid modified disposable carbon electrodes. Application to the determination of beta-lactam antibiotics Conzuelo, F. Gamella, María Campuzano, Susana Martínez-Ruiz, Paloma Esteban-Torres, María Rivas, Blanca de las Reviejo , A. J. Muñoz, Rosario Pingarrón, José M. Ministerio de Economía y Competitividad (España) Comunidad de Madrid Ministerio de Educación, Cultura y Deporte (España) A novel strategy for the construction of disposable amperometric affinity biosensors is described in this work. The approach uses a recombinant bacterial penicillin binding protein (PBP) tagged by an N-terminal hexahistidine tail which was immobilized onto Co2+−tetradentate nitrilotriacetic acid (NTA)-modified screen-printed carbon electrodes (SPCEs). The biosensor was employed for the specific detection and quantification of β-lactam antibiotics residues in milk, which was accomplished by means of a direct competitive assay using a tracer with horseradish peroxidase (HRP) for the enzymatic labeling. The amperometric response measured at −0.20 V versus the Ag pseudoreference electrode of the SPCE upon the addition of H2O2 in the presence of hydroquinone (HQ) as redox mediator was used as the transduction signal. The developed affinity sensor allowed limits of detection to be obtained in the low part-per-billion level for the antibiotics tested in untreated milk samples. Moreover, the biosensor exhibited a good selectivity against other antibiotics residues frequently detected in milk and dairy products. The analysis time was of approximately 30 min. The financial support of the Spanish Ministerio de Economía y Competitividad Research Projects, CTQ2012-34238, and the AVANSENS Program from the Comunidad de Madrid (S2009PPQ-1642) are gratefully acknowledged. Felipe Conzuelo acknowledges an FPU fellowship from the Spanish Ministry of Education. Peer Reviewed 2014-01-21T15:41:50Z 2014-01-21T15:41:50Z 2013 2014-01-21T15:41:50Z artículo http://purl.org/coar/resource_type/c_6501 issn: 0003-2700 Analytical Chemistry 85(6): 3246-3254 (2013) http://hdl.handle.net/10261/89711 10.1021/ac303604b http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100003176 http://dx.doi.org/10.13039/100012818 en #PLACEHOLDER_PARENT_METADATA_VALUE# S2009PPQ-164/AVANSENS Postprint http://dx.doi.org/10.1021/ac303604b Sí open American Chemical Society |
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A novel strategy for the construction of disposable amperometric affinity biosensors is described in this work. The approach uses a recombinant bacterial penicillin binding protein (PBP) tagged by an N-terminal hexahistidine tail which was immobilized onto Co2+−tetradentate nitrilotriacetic acid (NTA)-modified screen-printed carbon electrodes (SPCEs). The biosensor was employed for the specific detection and quantification of β-lactam antibiotics residues in milk, which was accomplished by means of a direct competitive assay using a tracer with horseradish peroxidase (HRP) for the enzymatic labeling. The amperometric response measured at −0.20 V versus the Ag pseudoreference electrode of the SPCE upon the addition of H2O2 in the presence of hydroquinone (HQ) as redox mediator was used as the transduction signal. The developed affinity sensor allowed limits of detection to be obtained in the low part-per-billion level for the antibiotics tested in untreated milk samples. Moreover, the biosensor exhibited a good selectivity against other antibiotics residues frequently detected in milk and dairy products. The analysis time was of approximately 30 min. |
author2 |
Ministerio de Economía y Competitividad (España) |
author_facet |
Ministerio de Economía y Competitividad (España) Conzuelo, F. Gamella, María Campuzano, Susana Martínez-Ruiz, Paloma Esteban-Torres, María Rivas, Blanca de las Reviejo , A. J. Muñoz, Rosario Pingarrón, José M. |
format |
artículo |
author |
Conzuelo, F. Gamella, María Campuzano, Susana Martínez-Ruiz, Paloma Esteban-Torres, María Rivas, Blanca de las Reviejo , A. J. Muñoz, Rosario Pingarrón, José M. |
spellingShingle |
Conzuelo, F. Gamella, María Campuzano, Susana Martínez-Ruiz, Paloma Esteban-Torres, María Rivas, Blanca de las Reviejo , A. J. Muñoz, Rosario Pingarrón, José M. Integrated amperometric affinity biosensors using Co2+-tetradentane nitrilotriacetic acid modified disposable carbon electrodes. Application to the determination of beta-lactam antibiotics |
author_sort |
Conzuelo, F. |
title |
Integrated amperometric affinity biosensors using Co2+-tetradentane nitrilotriacetic acid modified disposable carbon electrodes. Application to the determination of beta-lactam antibiotics |
title_short |
Integrated amperometric affinity biosensors using Co2+-tetradentane nitrilotriacetic acid modified disposable carbon electrodes. Application to the determination of beta-lactam antibiotics |
title_full |
Integrated amperometric affinity biosensors using Co2+-tetradentane nitrilotriacetic acid modified disposable carbon electrodes. Application to the determination of beta-lactam antibiotics |
title_fullStr |
Integrated amperometric affinity biosensors using Co2+-tetradentane nitrilotriacetic acid modified disposable carbon electrodes. Application to the determination of beta-lactam antibiotics |
title_full_unstemmed |
Integrated amperometric affinity biosensors using Co2+-tetradentane nitrilotriacetic acid modified disposable carbon electrodes. Application to the determination of beta-lactam antibiotics |
title_sort |
integrated amperometric affinity biosensors using co2+-tetradentane nitrilotriacetic acid modified disposable carbon electrodes. application to the determination of beta-lactam antibiotics |
publisher |
American Chemical Society |
publishDate |
2013 |
url |
http://hdl.handle.net/10261/89711 http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100003176 http://dx.doi.org/10.13039/100012818 |
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