Abstract
The development of two different immunoassays for the determination of fumonisin B1 in wheat samples is reported. A previously described mimopeptide for fumonisin B1 (FB1) was used to produce fusion proteins in combination with two different luciferases: Gaussia luciferase (GLuc) and NanoLuc luciferase (NLuc). The production, expression and the development of two immunoassays based on these fusion proteins (A2‐GLuc and A2‐NLuc) is detailed. The assay showing the best performance, A2‐NLuc, with a limit of detection of 0.61 ng mL−1 and a dynamic range from 1.9 to 95 ng mL−1, was employed for the analysis of spiked wheat samples, a reference matrix material, as well as naturally contaminated wheat samples. The recoveries obtained in the spiked samples were acceptable, between 81.5 and 109 %, with relative standard deviations lower than 14 %. The analysis of naturally contaminated wheat was validated by a liquid chromatography coupled to tandem mass detection method.
A comparative study of the performance of two different luciferases, Gaussia and NanoLuc Luciferase, expressed in combination with a fumonisin mimopeptide was carried out in this work. The fusion protein showing the best performance was used for the analysis of fumonisin B1 in wheat samples in combination with a bead‐based immunoassay.
