Repositori institucional URV
| URV's Author/s: | Jauset Rubio, Miriam / O'SULLIVAN, CIARA KATHLEEN / Ortíz Rodríguez, Mayreli |
| Author, as appears in the article.: | Jauset-Rubio, Miriam; Ortiz, Mayreli; O'Sullivan, Ciara K |
| Author's mail: | mayreli.ortiz@urv.cat miriam.jauset@urv.cat mayreli.ortiz@urv.cat |
| Author identifier: | 0000-0002-9423-0055 0000-0002-9943-6132 0000-0002-9423-0055 |
| Journal publication year: | 2022 |
| Publication Type: | Journal Publications |
| APA: | Jauset-Rubio, Miriam; Ortiz, Mayreli; O'Sullivan, Ciara K (2022). Exploiting the Nucleic Acid Nature of Aptamers for Signal Amplification. Biosensors, 12(11), 972-. DOI: 10.3390/bios12110972 |
| Papper original source: | Biosensors. 12 (11): 972- |
| Abstract: | Aptamer-based assays and sensors are garnering increasing interest as alternatives to antibodies, particularly due to their increased flexibility for implementation in alternative assay formats, as they can be employed in assays designed for nucleic acids, such as molecular aptamer beacons or aptamer detection combined with amplification. In this work, we took advantage of the inherent nucleic acid nature of aptamers to enhance sensitivity in a rapid and facile assay format. An aptamer selected against the anaphylactic allergen β-conglutin was used to demonstrate the proof of concept. The aptamer was generated by using biotinylated dUTPs, and the affinity of the modified aptamer as compared to the unmodified aptamer was determined by using surface plasmon resonance to calculate the dissociation constant (KD), and no significant improvement in affinity due to the incorporation of the hydrophobic biotin was observed. The modified aptamer was then applied in a colorimetric competitive enzyme-linked oligonucleotide assay, where β-conglutin was immobilized on the wells of a microtiter plate, competing with β-conglutin free in solution for the binding to the aptamer. The limit of detection achieved was 68 pM, demonstrating an improvement in detection limit of three orders of magnitude as compared with the aptamer simply modified with a terminal biotin label. The concept was then exploited by using electrochemical detection and screen-printed electrodes where detection limits of 326 fM and 7.89 fM were obtained with carbon and gold electrodes, respectively. The assay format is generic in nature and can be applied to all aptamers, facilitating an easy and cost-effective means to achieve lower detection limits. |
| Article's DOI: | 10.3390/bios12110972 |
| Link to the original source: | https://www.mdpi.com/2079-6374/12/11/972 |
| Papper version: | info:eu-repo/semantics/publishedVersion |
| licence for use: | https://creativecommons.org/licenses/by/3.0/es/ |
| Department: | Enginyeria Química |
| Licence document URL: | https://repositori.urv.cat/ca/proteccio-de-dades/ |
| Thematic Areas: | Química Nanoscience & nanotechnology Medicine (miscellaneous) Materiais Instruments & instrumentation Instrumentation Engineering (miscellaneous) Clinical biochemistry Ciencias sociales Ciências biológicas iii Ciências biológicas i Chemistry, analytical Biotecnología Biotechnology Biomedical engineering Astronomia / física Analytical chemistry |
| Keywords: | High-affinity Enzyme-linked oligonucleotide assay (elona) Electrochemical detection Chronoamperometry Biotinylated dntps Aptasensors Aptamer rna probes nucleotides generation enzyme-linked oligonucleotide assay (elona) electrochemical detection dntps dna chronoamperometry biotinylated dntps binding avidin aptasensors |
| Entity: | Universitat Rovira i Virgili |
| Record's date: | 2024-10-26 |
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