Bioluminescent detection of zearalenone using recombinant peptidomimetic Gaussia luciferase fusion protein

使用重组肽模拟高斯荧光素酶融合蛋白进行玉米赤霉烯酮的生物发光检测

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作者:Riikka Peltomaa, Sabrina Fikacek, Elena Benito-Peña, Rodrigo Barderas, Trajen Head, Sapna Deo, Sylvia Daunert, María C Moreno-Bondi

Abstract

The development of a bioluminescent immunosensor is reported for the determination of zearalenone (ZEA) based on a peptide mimetic identified by phage display. The peptide mimetic GW, with a peptide sequence GWWGPYGEIELL, was used to create recombinant fusion proteins with the bioluminescent Gaussia luciferase (GLuc) that were directly used as tracers for toxin detection in a competitive immunoassay without the need for secondary antibodies or further labeling. The bioluminescent sensor, based on protein G-coupled magnetic beads for antibody immobilization, enabled determination of ZEA with a detection limit of 4.2 ng mL-1 (corresponding to 420 μg kg-1 in food samples) and an IC50 value of 11.0 ng mL-1. The sensor performance was evaluated in spiked maize and wheat samples, with recoveries ranging from 87 to 106% (RSD < 20%, n = 3). Finally, the developed method was applied to the analysis of a naturally contaminated reference matrix material and good agreement with the reported concentrations was obtained.Graphical abstract.

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