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Split Aptamers Immobilized on Polymer Brushes Integrated in a Lab-on-Chip System Based on an Array of Amorphous Silicon Photosensors: A Novel Sensor Assay

Innovative materials for the integration of aptamers in Lab-on-Chip systems are important for the development of miniaturized portable devices in the field of health-care and diagnostics. Herein we highlight a general method to tailor an aptamer sequence in two subunits that are randomly immobilized...

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Autores principales: Nandimandalam, Manasa, Costantini, Francesca, Lovecchio, Nicola, Iannascoli, Lorenzo, Nascetti, Augusto, de Cesare, Giampiero, Caputo, Domenico, Manetti, Cesare
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8658169/
https://www.ncbi.nlm.nih.gov/pubmed/34885364
http://dx.doi.org/10.3390/ma14237210
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author Nandimandalam, Manasa
Costantini, Francesca
Lovecchio, Nicola
Iannascoli, Lorenzo
Nascetti, Augusto
de Cesare, Giampiero
Caputo, Domenico
Manetti, Cesare
author_facet Nandimandalam, Manasa
Costantini, Francesca
Lovecchio, Nicola
Iannascoli, Lorenzo
Nascetti, Augusto
de Cesare, Giampiero
Caputo, Domenico
Manetti, Cesare
author_sort Nandimandalam, Manasa
collection PubMed
description Innovative materials for the integration of aptamers in Lab-on-Chip systems are important for the development of miniaturized portable devices in the field of health-care and diagnostics. Herein we highlight a general method to tailor an aptamer sequence in two subunits that are randomly immobilized into a layer of polymer brushes grown on the internal surface of microfluidic channels, optically aligned with an array of amorphous silicon photosensors for the detection of fluorescence. Our approach relies on the use of split aptamer sequences maintaining their binding affinity to the target molecule. After binding the target molecule, the fragments, separately immobilized to the brush layer, form an assembled structure that in presence of a “light switching” complex [Ru(phen)(2)(dppz)](2+), emit a fluorescent signal detected by the photosensors positioned underneath. The fluorescent intensity is proportional to the concentration of the target molecule. As proof of principle, we selected fragments derived from an aptamer sequence with binding affinity towards ATP. Using this assay, a limit of detection down to 0.9 µM ATP has been achieved. The sensitivity is compared with an assay where the original aptamer sequence is used. The possibility to re-use both the aptamer assays for several times is demonstrated.
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spelling pubmed-86581692021-12-10 Split Aptamers Immobilized on Polymer Brushes Integrated in a Lab-on-Chip System Based on an Array of Amorphous Silicon Photosensors: A Novel Sensor Assay Nandimandalam, Manasa Costantini, Francesca Lovecchio, Nicola Iannascoli, Lorenzo Nascetti, Augusto de Cesare, Giampiero Caputo, Domenico Manetti, Cesare Materials (Basel) Article Innovative materials for the integration of aptamers in Lab-on-Chip systems are important for the development of miniaturized portable devices in the field of health-care and diagnostics. Herein we highlight a general method to tailor an aptamer sequence in two subunits that are randomly immobilized into a layer of polymer brushes grown on the internal surface of microfluidic channels, optically aligned with an array of amorphous silicon photosensors for the detection of fluorescence. Our approach relies on the use of split aptamer sequences maintaining their binding affinity to the target molecule. After binding the target molecule, the fragments, separately immobilized to the brush layer, form an assembled structure that in presence of a “light switching” complex [Ru(phen)(2)(dppz)](2+), emit a fluorescent signal detected by the photosensors positioned underneath. The fluorescent intensity is proportional to the concentration of the target molecule. As proof of principle, we selected fragments derived from an aptamer sequence with binding affinity towards ATP. Using this assay, a limit of detection down to 0.9 µM ATP has been achieved. The sensitivity is compared with an assay where the original aptamer sequence is used. The possibility to re-use both the aptamer assays for several times is demonstrated. MDPI 2021-11-26 /pmc/articles/PMC8658169/ /pubmed/34885364 http://dx.doi.org/10.3390/ma14237210 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Nandimandalam, Manasa
Costantini, Francesca
Lovecchio, Nicola
Iannascoli, Lorenzo
Nascetti, Augusto
de Cesare, Giampiero
Caputo, Domenico
Manetti, Cesare
Split Aptamers Immobilized on Polymer Brushes Integrated in a Lab-on-Chip System Based on an Array of Amorphous Silicon Photosensors: A Novel Sensor Assay
title Split Aptamers Immobilized on Polymer Brushes Integrated in a Lab-on-Chip System Based on an Array of Amorphous Silicon Photosensors: A Novel Sensor Assay
title_full Split Aptamers Immobilized on Polymer Brushes Integrated in a Lab-on-Chip System Based on an Array of Amorphous Silicon Photosensors: A Novel Sensor Assay
title_fullStr Split Aptamers Immobilized on Polymer Brushes Integrated in a Lab-on-Chip System Based on an Array of Amorphous Silicon Photosensors: A Novel Sensor Assay
title_full_unstemmed Split Aptamers Immobilized on Polymer Brushes Integrated in a Lab-on-Chip System Based on an Array of Amorphous Silicon Photosensors: A Novel Sensor Assay
title_short Split Aptamers Immobilized on Polymer Brushes Integrated in a Lab-on-Chip System Based on an Array of Amorphous Silicon Photosensors: A Novel Sensor Assay
title_sort split aptamers immobilized on polymer brushes integrated in a lab-on-chip system based on an array of amorphous silicon photosensors: a novel sensor assay
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8658169/
https://www.ncbi.nlm.nih.gov/pubmed/34885364
http://dx.doi.org/10.3390/ma14237210
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