Competitive upconversion-linked immunoassay using peptide mimetics for the detection of the mycotoxin zearalenone

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Authors	PELTOMAA Riikka FARKA Zdeněk MICKERT Matthias Jürgen BRANDMEIER Julian PASTUCHA Matěj HLAVÁČEK Antonín MARTÍNEZ-ORTS Mónica CANALES Ángeles SKLÁDAL Petr BENITO-PENA Elena MORENO-BONDI María C. GORRIS Hans-Heiner
Year of publication	2020
Type	Article in Periodical
Magazine / Source	Biosensors and Bioelectronics
MU Faculty or unit	Faculty of Science
Citation
Web	https://www.sciencedirect.com/science/article/pii/S0956566320306722
Doi	http://dx.doi.org/10.1016/j.bios.2020.112683
Keywords	Zearalenone; Upconversion nanoparticle; Peptide mimetic; Immunosensing; Surface plasmon resonance; Food safety
Description	Due to increasing food safety standards, the analysis of mycotoxins has become essential in the food industry. In this work, we have developed a competitive upconversion-linked immunosorbent assay (ULISA) for the analysis of zearalenone (ZEA), one of the most frequently encountered mycotoxins in food worldwide. Instead of a toxin-conjugate conventionally used in competitive immunoassays, we designed a ZEA mimicking peptide extended by a biotin-linker and confirmed its excellent suitability to mimic ZEA by nuclear magnetic resonance (NMR) and surface plasmon resonance (SPR) analysis. Upconversion nanoparticles (UCNP, type NaYF4:Yb,Tm) served as background-free optical label for the detection of the peptide mimetic in the competitive ULISA. Streptavidin-conjugated UCNPs were prepared by click reaction using an alkyne-PEG-neridronate linker. The UCNP conjugate clearly outperformed conventional labels such as enzymes or fluorescent dyes. With a limit of detection of 20 pg mL-1 (63 pM), the competitive ULISA is well applicable to the detection of ZEA at the levels set by the European legislation. Moreover, the ULISA is specific for ZEA and its metabolites (alpha- and beta-zearalenol) without significant cross-reactivity with other related mycotoxins. We detected ZEA in spiked and naturally contaminated maize samples using liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) as a reference method to demonstrate food analysis in real samples.
Related projects:	CEITEC 2020 Sensitive bioanalytical tools for the surveillance of honey bee diseases Czech Infrastructure for Integrative Structural Biology Podpora biochemického výzkumu v roce 2020