Development of a recombinant Fab-fragment based electrochemical immunosensor for deoxynivalenol detection in food samples

Abstract

a b s t r a c tA reliable and cost-effective electrochemical method for the detection of deoxynivalenol (DON) in cereals and cereal-based food samples based on the use of a novel anti-DON Fab fragment is presented. The analytical system employed, Enzyme-Linked-Immunomagnetic-Electrochemical (ELIME) assay, is based on the use of immunomagnetic beads (IMBs) coupled with eight magnetized screen-printed electrodes (8-mScPEs) as electrochemical transducers.Using standard solutions of DON, a working range between 100 and 4500 ng/ml was obtained with an EC 50 of 380 ng/ml. The ELIME assay was employed to evaluate the cross-reactivity of the Fab fragment towards different trichothecenes revealing a good selectivity towards DON over other trichothecenes with the exception of 3-Ac-DON. The sensor was then applied to cereals and cereal-based food samples (wheat, breakfast cereal and baby-food) and a wide range of sample treatment procedures was tested. Within-laboratory precision (9-24% repeatability for breakfast cereals and 10-33% for baby-food) and recovery data (82-110% for breakfast cereals and 97-108% for baby-food) were calculated by analyzing blank breakfast cereals and baby-foods fortified with DON, demonstrating that the proposed method has the capability for use as a screening assay for DON in such products.

Development of an immunochromatographic strip test for the rapid detection of zearalenone in wheat from Jiangsu province, China

“…Gradient dilutions were prepared with stock extraction solution and the 2-fold dilution was selected finally due to smaller effect on matrix and higher sensitivity to ZEN. In addition, it was demonstrated in previous studies that the extraction solution of acetonitrile and water co-extracted less interfering compounds in matrix compared with the solvent of methanol and water thus the former was used in this study [28, 29]. …”

Magnetic Beads‐Based Electrochemical Sensors Applied to the Detection and Quantification of Bioterrorism/Biohazard Agents

“…A related approach has been described for the detection of deoxynivalenol (DON), a mycotoxin produced by Fusarium fungi which represents a serious threat for the safety of cereal-based food and feedstuffs [34]. This methodology was based on the use of a novel anti-DON Fab fragment and of immunomagnetic beads coupled with eight magnetized SPEs as electrochemical transducers.…”

Mycotoxin Determination in Foods Using Advanced Sensors Based on Antibodies or Aptamers

“…Based on the use of immunomagnetic beads (IMBs) coupled with recombinant anti-DON Fab fragment, Romanazzo [94] developed an enzyme-linked-immunomagnetic-electrochemical (ELIME) method for DON detection in cereal and cereal-based food samples. The recombinant anti-DON Fab fragment was used as recognition element and the eight magnetized screen-printed electrodes were employed as electrochemical transducers.…”