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dc.creatorCiáurriz Gortari, Paulaes_ES
dc.creatorFernández, Fátimaes_ES
dc.creatorTellechea, Edurnees_ES
dc.creatorMorán Juez, José Fernandoes_ES
dc.creatorAsensio, Aarón C.es_ES
dc.date.accessioned2018-08-31T08:07:56Z
dc.date.available2018-08-31T08:07:56Z
dc.date.issued2017
dc.identifier.issn2190-4286
dc.identifier.urihttps://hdl.handle.net/2454/30345
dc.description.abstractThe enzyme-linked immunosorbent assay (ELISA) technique is based on the specific recognition ability of the molecular structure of an antigen (epitope) by an antibody and is likely the most important diagnostic technique used today in bioscience. With this methodology, it is possible to diagnose illness, allergies, alimentary fraud, and even to detect small molecules such as toxins, pesticides, heavy metals, etc. For this reason, any procedures that improve the detection limit, sensitivity or reduce the analysis time could have an important impact in several fields. In this respect, many methods have been developed for improving the technique, ranging from fluorescence substrates to methods for increasing the number of enzyme molecules involved in the detection such as the biotin–streptavidin method. In this context, nanotechnology has offered a significant number of proposed solutions, mainly based on the functionalization of nanoparticles from gold to carbon which could be used as antibody carriers as well as reporter enzymes like peroxidase. However, few works have focused on the study of best practices for nanoparticle functionalization for ELISA enhancement. In this work, we use 20 nm gold nanoparticles (AuNPs) as a vehicle for secondary antibodies and peroxidase (HRP). The design of experiments technique (DOE) and four different methods for biomolecule loading were compared using a rabbit IgG/goat anti-rabbit IgG ELISA model (adsorption, directional, covalent and a combination thereof). As a result, AuNP probes prepared by direct adsorption were the most effective method. AuNPs probes were then used to detect gliadin, one of the main components of wheat gluten, the protein composite that causes celiac disease. With this optimized approach, our data showed a sensitivity increase of at least five times and a lower detection limit with respect to a standard ELISA of at least three times. Additionally, the assay time was remarkably decreased.en
dc.description.sponsorshipThe authors would like to thank the Government of Navarra, Department of Innovation, Business and Employment for financial support within the project SABioD.en
dc.format.extent10 p.
dc.format.mimetypeapplication/pdfen
dc.format.mimetypeapplication/zipen
dc.language.isoengen
dc.publisherBeilstein-Instituten
dc.relation.ispartofBeilstein Journal of Nanotechnology, 2017, 8, 244–253en
dc.rights© 2017 Ciaurriz et al.; licensee Beilstein-Institut. This is an Open Access article under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The license is subject to the Beilstein Journal of Nanotechnology terms and conditions: (http://www.beilstein-journals.org/bjnano)en
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectAllergenen
dc.subjectELISA enhancementen
dc.subjectFunctionalizationen
dc.subjectGliadinen
dc.subjectGold nanoparticleen
dc.titleComparison of four functionalization methods of gold nanoparticles for enhancing the enzyme-linked immunosorbent assay (ELISA)en
dc.typeinfo:eu-repo/semantics/articleen
dc.typeArtículo / Artikuluaes
dc.contributor.departmentIdAB - Instituto de Agrobiotecnología / Agrobioteknologiako Institutuaes
dc.rights.accessRightsinfo:eu-repo/semantics/openAccessen
dc.rights.accessRightsAcceso abierto / Sarbide irekiaes
dc.identifier.doi10.3762/bjnano.8.27
dc.relation.publisherversionhttps://doi.org/10.3762/bjnano.8.27
dc.type.versioninfo:eu-repo/semantics/publishedVersionen
dc.type.versionVersión publicada / Argitaratu den bertsioaes
dc.contributor.funderGobierno de Navarra / Nafarroako Gobernua, proyecto SABioDes


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© 2017 Ciaurriz et al.; licensee Beilstein-Institut. This is an Open Access article under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The license is subject to the Beilstein Journal of Nanotechnology terms and conditions: (http://www.beilstein-journals.org/bjnano)
Except where otherwise noted, this item's license is described as © 2017 Ciaurriz et al.; licensee Beilstein-Institut. This is an Open Access article under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The license is subject to the Beilstein Journal of Nanotechnology terms and conditions: (http://www.beilstein-journals.org/bjnano)