Comparison of four functionalization methods of gold nanoparticles for enhancing the enzyme-linked immunosorbent assay (ELISA)
Fecha
2017Autor
Versión
Acceso abierto / Sarbide irekia
Tipo
Artículo / Artikulua
Versión
Versión publicada / Argitaratu den bertsioa
Impacto
|
10.3762/bjnano.8.27
Resumen
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, ...
[++]
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. [--]
Materias
Allergen,
ELISA enhancement,
Functionalization,
Gliadin,
Gold nanoparticle
Editor
Beilstein-Institut
Publicado en
Beilstein Journal of Nanotechnology, 2017, 8, 244–253
Departamento
Universidad Pública de Navarra/Nafarroako Unibertsitate Publikoa. IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua
Versión del editor
Entidades Financiadoras
The authors would like to thank the Government of Navarra, Department of Innovation, Business and Employment for financial support within the project SABioD.
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La licencia del ítem se describe como © 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)