Bioluminescent detection of zearalenone using recombinant peptidomimetic Gaussia luciferase fusion protein
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ORIGINAL PAPER
Bioluminescent detection of zearalenone using recombinant peptidomimetic Gaussia luciferase fusion protein Riikka Peltomaa 1 & Sabrina Fikacek 1 & Elena Benito-Peña 1 Sylvia Daunert 3,4,5 & María C. Moreno-Bondi 1
&
Rodrigo Barderas 2 & Trajen Head 3,4 & Sapna Deo 3,4 &
Received: 23 May 2020 / Accepted: 28 August 2020 # Springer-Verlag GmbH Austria, part of Springer Nature 2020
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. Keywords Bioluminescence . Gaussia luciferase . Mycotoxin . Zearalenone . Food safety
Introduction Mycotoxins, which are commonly encountered as food and feed contaminants, are secondary metabolites produced by
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00604-020-04538-7) contains supplementary material, which is available to authorized users. * Elena Benito-Peña [email protected] * María C. Moreno-Bondi [email protected] 1
Department of Analytical Chemistry, Faculty of Chemistry, Complutense University, Ciudad Universitaria s/n, 28040 Madrid, Spain
2
Chronic Disease Programme, UFIEC, Instituto de Salud Carlos III, Ctra. Majadahonda-Pozuelo Km 2.2, 28220 Madrid, Spain
3
Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL 33136, USA
4
Dr. JT Macdonald Foundation Biomedical Nanotechnology Institute, University of Miami, Coral Gables, FL 33136, USA
5
University of Miami Clinical and Translational Science Institute, University of Miami, Miami, FL 33136, USA
filamentous fungi. These toxins have a wide range of harmful effects of the neurotoxic, carcinogenic, and immunotoxic types on humans and other vertebrates; also, they alter their development and reproductive functions [1–3]. Aflatoxins, ochratoxins, fumonisins, trichothecenes, and zearalenone are among the major mycotoxins produced by ubiquitous fungal species [3, 4]. Zearalenone (ZEA) is a nonsteroidal estrogenic mycotoxin produced by various Fusarium species but particularly F. graminearum. By virtue of their similarity to 17βestradiol, ZEA and
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