Abstract: A strategy to design an immunoassay based on imprinted proteins to detect a foodborne toxin zearalenone (ZEN) has been proposed. Proteins were used as scaffolds for generation of binding cavities with a high specificity against ZEN. Different proteins and different bioimprinting approaches were tested. The imprinted proteins primarily prepared in a tube and then immobilized on a microwell plate, and directly prepared in the microwell plate were compared for an application in immunosorbent detection of ZEN in naturally contaminated wheat and maize samples. The assay was combined with a rapid extraction technique. Specific interactions between the imprinted proteins and the target in a μg kg-1 linear range was achieved. As a confirmatory method, liquid chromatography coupled to tandem mass spectrometry was used. The obtained results allow using the developed immunoassay for the analysis of ZEN with a sensitivity matching the maximum permitted level for ZEN in unprocessed cereals established by the European Commission (100 μg kg-1)
Author keywords: Imprinted protein, Bioimprinting, ELISA, Zearalenone, rapid screening, mycotoxin