Comparison of four functionalization methods of gold nanoparticles for enhancing the enzyme-linked immunosorbent assay (ELISA)

Ciáurriz Gortari, Paula; Fernández Santos, Fátima; Tellechea Malda, Edurne; Morán Juez, José Fernando; Asensio, Aarón C.

Comparison of four functionalization methods of gold nanoparticles for enhancing the enzyme-linked immunosorbent assay (ELISA)

dc.contributor.author	Ciáurriz Gortari, Paula
dc.contributor.author	Fernández Santos, Fátima
dc.contributor.author	Tellechea Malda, Edurne
dc.contributor.author	Morán Juez, José Fernando
dc.contributor.author	Asensio, Aarón C.
dc.contributor.department	IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua	es_ES
dc.contributor.funder	Gobierno de Navarra / Nafarroako Gobernua, proyecto SABioD	es
dc.date.accessioned	2018-08-31T08:07:56Z
dc.date.available	2018-08-31T08:07:56Z
dc.date.issued	2017
dc.description.abstract	The 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.sponsorship	The 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.extent	10 p.
dc.format.mimetype	application/pdf	en
dc.format.mimetype	application/zip	en
dc.identifier.doi	10.3762/bjnano.8.27
dc.identifier.issn	2190-4286
dc.identifier.uri	https://academica-e.unavarra.es/handle/2454/30345
dc.language.iso	eng	en
dc.publisher	Beilstein-Institut	en
dc.relation.ispartof	Beilstein Journal of Nanotechnology, 2017, 8, 244–253	en
dc.relation.publisherversion	https://doi.org/10.3762/bjnano.8.27
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.accessRights	info:eu-repo/semantics/openAccess
dc.rights.uri	https://creativecommons.org/licenses/by/4.0/
dc.subject	Allergen	en
dc.subject	ELISA enhancement	en
dc.subject	Functionalization	en
dc.subject	Gliadin	en
dc.subject	Gold nanoparticle	en
dc.title	Comparison of four functionalization methods of gold nanoparticles for enhancing the enzyme-linked immunosorbent assay (ELISA)	en
dc.type	info:eu-repo/semantics/article
dc.type.version	info:eu-repo/semantics/publishedVersion
dspace.entity.type	Publication
relation.isAuthorOfPublication	cad13109-2660-4ec4-80c8-e4bcc0685382
relation.isAuthorOfPublication.latestForDiscovery	cad13109-2660-4ec4-80c8-e4bcc0685382