title: Nanogold bioconjugates for direct and sensitive multiplexed immunosensing.
abstract: The use of nanogold bioconjugates for direct detection of the antibody-antigen immunoreaction is addressed. The integration of gold nanoparticles tracers as signal generators in microarray immunosensing and compact disc detection technique show important advantages to reach sensitive, selective, high throughput, reliable and cost-effective assays. For that, a thorough study of the performances of the size of spherical nanogold particles and coating density was developed. The size of the nanoparticle determines the optimal antibody dilution, being the smaller particles the best performing ones. Enhancement effect of lower size is also studied. The gold labeling method do not affects the recognition capability of the labeled proteins. As a proof of concept, the nanoconjugates were used for the simultaneous and direct determination of small molecules. Employing nanogold bioconjugates as recognition labels resulted in robust and reliable assays, reaching a sensitivity of 0.03 and 1.3μg/L for sulfasalazine and atrazine, respectively. This shows that the use of nanogold bioconjugates for direct immunosensing is very competitive, achieving highly sensitive and reproducible assays (RSD<10%). This approach would simultaneously determine both small and large molecular size targets, in different formats, using the same detection mode what paves the way for many other applications in different scenarios.
authors.affiliation: Instituto Interuniversitario de Reconocimiento Molecular y Desarrollo Tecnológico (IDM), Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46071 Valencia, Spain.
authors.forename: P
authors.initials: P
authors.lastname: Dobosz
authors.affiliation: Instituto Interuniversitario de Reconocimiento Molecular y Desarrollo Tecnológico (IDM), Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46071 Valencia, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Morais
authors.affiliation: Instituto Interuniversitario de Reconocimiento Molecular y Desarrollo Tecnológico (IDM), Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46071 Valencia, Spain.
authors.forename: R
authors.initials: R
authors.lastname: Puchades
authors.affiliation: Instituto Interuniversitario de Reconocimiento Molecular y Desarrollo Tecnológico (IDM), Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46071 Valencia, Spain. Electronic address: amaquieira@qim.upv.es.
authors.forename: A
authors.initials: A
authors.lastname: Maquieira
mesh_terms: D016250:Compact Disks; D045424:Complex Mixtures / Q000032:analysis*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006046:Gold / Q000737:chemistry*; D007118:Immunoassay / Q000295:instrumentation*; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D046228:Microarray Analysis / Q000295:instrumentation*; D058726:Nanoconjugates / Q000737:chemistry / Q000648:ultrastructure
chemical_list: D045424:Complex Mixtures; D058726:Nanoconjugates; D006046:Gold
keywords: Immunoassay; Microarraying; Multiplex; Nano-gold; Tracers