A1 Refereed original research article in a scientific journal
Bioluminescent detection of zearalenone using recombinant peptidomimetic Gaussia luciferase fusion protein
Authors: Peltomaa R, Fikacek S, Benito-Pena E, Barderas R, Head T, Deo S, Daunert S, Moreno-Bondi MC
Publisher: Springer
Publication year: 2020
Journal:Microchimica Acta
Journal name in sourceMICROCHIMICA ACTA
Journal acronym: Microchimica Acta
Article number: 547
Volume: 187
Issue: 10
Number of pages: 11
ISSN: 0026-3672
DOI: https://doi.org/10.1007/s00604-020-04538-7
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 mu g 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.
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 mu g 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.
UTU Research Portal