RT Journal Article
T1 Homogeneous quenching immunoassay for fumonisin B1 based on gold nanoparticles and an epitope-mimicking yellow fluorescent protein
A1 Peltomaa, Riikka Johanna
A1 Amaro Torres, Francisco
A1 Carrasco, Sergio
A1 Orellana Moraleda, Guillermo
A1 Benito Peña, María Elena
A1 Moreno Bondi, María Cruz
AB Homogeneous immunoassays represent an attractive alternative to traditional heterogeneous assays due to their simplicity, sensitivity, and speed. On the basis of a previously identified epitope-mimicking peptide, or mimotope, we developed a homogeneous fluorescence quenching immunoassay based on gold nanoparticles (AuNPs) and a recombinant epitope-mimicking fusion protein for the detection of mycotoxin fumonisin B1 (FB1). The fumonisin mimotope was cloned as a fusion protein with a yellow fluorescent protein that could be used directly as the tracer for FB1 detection without the need of labeling or a secondary antibody. Furthermore, owing to the fluorescence quenching ability of AuNPs, a homogeneous immunoassay could be performed in a single step without washing steps to separate the unbound tracer. The homogeneous quenching assay showed negligible matrix effects in 5% wheat extract and high sensitivity for FB1 detection, with a dynamic range from 7.3 to 22.6 ng/mL, a detection limit of 1.1 ng/mL, and IC50 value of 12.9 ng/mL, which was significantly lower than the IC50 value of the previously reported assay using the synthetic counterpart of the same mimotope in a microarray format. The homogeneous assay was demonstrated to be specific for fumonisins B1 and B2, as no significant cross-reactivity with other mycotoxins was observed, and acceptable recoveries (86% for FB1 2000 μg/kg and 103% for FB1 4000 μg/kg), with relative standard deviation less than 6.5%, were reported from spiked wheat samples, proving that the method could provide a valuable tool for simple analysis of mycotoxin-contaminated food samples.
PB American Chemical Society
SN 1936-0851
YR 2018
FD 2018
LK https://hdl.handle.net/20.500.14352/93316
UL https://hdl.handle.net/20.500.14352/93316
LA eng
NO Riikka Peltomaa, Francisco Amaro-Torres, Sergio Carrasco, Guillermo Orellana, Elena Benito-Peña, and María C. Moreno-BondiACS Nano 2018 12 (11), 11333-11342DOI: 10.1021/acsnano.8b06094
NO Universidad Complutense de Madrid
NO Ministerio de Economía y Competitividad (España)
DS Docta Complutense
RD 30 sept 2024