Repositori institucional URV
| Autor/es de la URV: | NADAL POLO, PEDRO / O'SULLIVAN, CIARA KATHLEEN / Pinto Vivas, Amanda Magdalena / PINTO, ALESSANDRO / SVOBODOVÁ, MARKÉTA |
| Autor según el artículo: | Nadal, Pedro; Pinto, Alessandro; Svobodova, Marketa; Canela, Nuria; O'Sullivan, Ciara K |
| Direcció de correo del autor: | amanda.pinto@estudiants.urv.cat |
| Año de publicación de la revista: | 2012 |
| Tipo de publicación: | Journal Publications |
| Referencia de l'ítem segons les normes APA: | Nadal, Pedro; Pinto, Alessandro; Svobodova, Marketa; Canela, Nuria; O'Sullivan, Ciara K (2012). DNA aptamers against the lup an 1 food allergen. Plos One, 7(4), e35253-e35253. DOI: 10.1371/journal.pone.0035253 |
| Referencia al articulo segun fuente origial: | Plos One. 7 (4): e35253-e35253 |
| Resumen: | Using in vitro selection, high affinity DNA aptamers to the food allergen Lup an 1, ß-conglutin, were selected from a pool of DNA, 93 bases in length, containing a randomised sequence of 49 bases. ß-conglutin was purified from lupin flour and chemically crosslinked to carboxylated magnetic beads. Peptide mass fingerprinting was used to confirm the presence of the ß-conglutin. Single stranded DNA was generated from the randomised pool using T7 Gene 6 Exonuclease and was subsequently incubated with the magnetic beads and the captured DNA was released and amplified prior to a further round of Systematic Evolution of Ligands by Exponential Enrichment (SELEX). Evolution was monitored using enzyme linked oligonucleotide assay and surface plasmon resonance. Once a plateau in evolution was reached, the isolated DNA sequences were cloned and sequenced. The consensus motif was identified via alignment of the sequences and the affinities of these sequences for immobilised ß-conglutin were determined using surface plasmon resonance. The selected aptamer was demonstrated to be highly specific, showing no cross-reactivity with other flour ingredients or with other conglutin fractions of lupin. The secondary structures of the selected aptamers were predicted using m-fold. Finally, the functionality of the selected aptamers was demonstrated using a competitive assay for the quantitative detection of ß-conglutin. Future work will focus on structure elucidation and truncation of the selected sequences to generate a smaller aptamer for application to the analysis of the Lup an 1 allergen in foodstuffs. © 2012 Nadal et al. |
| DOI del artículo: | 10.1371/journal.pone.0035253 |
| Enlace a la fuente original: | https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0035253 |
| Versión del articulo depositado: | info:eu-repo/semantics/publishedVersion |
| Acceso a la licencia de uso: | https://creativecommons.org/licenses/by/3.0/es/ |
| Departamento: | Enginyeria Química |
| URL Documento de licencia: | https://repositori.urv.cat/ca/proteccio-de-dades/ |
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| Entidad: | Universitat Rovira i Virgili |
| Fecha de alta del registro: | 2024-11-16 |
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