Development of a monoclonal antibody against deoxynivalenol for magnetic nanoparticle-based extraction and an enzyme-linked immunosorbent assay

Abstract

Monoclonal antibody (mAb, NVRQS-DON) against deoxynivalenol (DON) was prepared. DON-Ag coated enzyme linked immunosorbent assay (ELISA) and DON-Ab coated ELISA were prepared by coating the DON-BSA and DON mAb. Quantitative DON calculation ranged from 50 to 4,000 ng/mL for DON-Ab coated ELISA and from 25 to 500 ng/mL for DON-Ag coated ELISA. 50% of inhibitory concentration values of DON, HT-2, 15-acetyl-DON, and nivalenol were 23.44, 22,545, 5,518 and 5,976 ng/mL based on the DON-Ab coated ELISA. Cross-reactivity levels of the mAb to HT-2, 15-acetyl-DON, and nivalenol were 0.1, 0.42, and 0.40%. The intra- and interassay precision coefficient variation (CV) were both <10%. In the mAb-coated ELISA, mean DON recovery rates in animal feed (0 to 1,000 µg/kg) ranged from 68.34 to 95.49% (CV; 4.10 to 13.38%). DON in a buffer solution (250, 500 and 1,000 ng/mL) was isolated using 300 µg of NVRQS-DON and 3 mg of magnetic nanoparticles (MNPs). The mean recovery rates of DON using this mAb-MNP system were 75.2, 96.9, and 88.1% in a buffer solution spiked with DON (250, 500, and 1,000 ng/mL). Conclusively we developed competitive ELISAs for detecting DON in animal feed and created a new tool for DON extraction using mAb-coupled MNPs.

Risks to human and animal health related to the presence of deoxynivalenol and its acetylated and modified forms in food and feed

“…Different antibodies against DON are (commercially) available and described in the literature (Usleber et al., ; Maragos and McCormick, ; Maragos et al., ; Lee et al., ; Sanders et al., ). Antibodies against DON can cross‐react with acetylated and/or modified forms of DON.…”

Selective Nanoparticles in Microextraction

“…This scheme has been followed for the development of monoclonal Abs (mAbs) against deaxynivalenol, a mycotoxin. (29) These mAbs are used not only for the ELISA determination of this compound but also for the development of MNPs able to extract it form a liquid media. Recovery values, in the range from 75.2% to 96.9%, indicate the high potential of this procedure.…”

Rapid determination of aflatoxin B1 by an automated immunomagnetic bead purification sample pretreatment method combined with high‐performance liquid chromatography

“…In recent years, magnetic nanoparticles based technology with multidisciplinary and remarkable characteristics offers great possibilities for development for new analytical methodology [24–26]. Immunomagnetic beads (IMBs) due to the distinct physical and chemical properties, such as uniform diameter particles, large surface area, simplicity, superior compatibility, good sensitivity, and easy operation that can be homogenously distributed in colloidal suspension, have been widely applied in various fields, including nucleic acid extraction, peptide and protein analysis, and antibody purification [27–31].…”