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Rapid surface plasmon resonance-based inhibition assay of deoxynivalenol
Anna J Tüdös1, Elly R Lucas-van den Bos, Edwin C A Stigter
1Processing Quality & Safety, NIZO Food Research, Kernhemseweg 2, 6718 ZB Ede, The Netherlands. anna.tudos@NIZO.nl
Abstract:
Deoxynivalenol belongs to a group of highly toxic fungal metabolites produced by Fusarium species that may contaminate food and animal feed, mostly grains. Three different monoclonal mouse anti-deoxynivalenol antibodies were compared for the development of a surface plasmon resonance (SPR)-based immunoassay for the selective and quantitative determination of deoxynivalenol in naturally contaminated matrices. A conjugate of deoxynivalenol with the protein casein was prepared and immobilized on the sensor chip surface. An excess of antibody was added to each test solution before the measurement. The assay was based on the competition for antibody binding between the immobilized deoxynivalenol conjugate on the sensor and the free deoxynivalenol molecules in the test solution. The deoxynivalenol-casein sensor could be reused more than 500 times without significant loss of activity using 6 M guanidine chloride solution for regeneration. The cross-reactivity of the three antibodies in the SPR assay was tested with other trichothecene mycotoxins (3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, nivalenol, HT2-toxin, and T2-toxin). The only sample preparation was extraction with max 80 vol % acetonitrile and 10-fold dilution with the running buffer. The assay had an optimal range between 2.5 and 30 ng/mL deoxynivalenol in the test solution. Most results of the SPR-based assay were in agreement with liquid chromatography/tandem mass spectrometry measurements of naturally contaminated wheat samples.
Insights
A new surface plasmon resonance (SPR) immunoassay was developed for detecting deoxynivalenol (DON) in food and feed. This sensitive and reusable assay accurately quantifies DON in contaminated samples.
Area of Science:
- Food Science
- Analytical Chemistry
- Immunotechnology
Background:
- Deoxynivalenol (DON) is a toxic fungal metabolite contaminating grains, posing risks to food and animal feed.
- Accurate detection methods are crucial for monitoring DON contamination in agricultural products.
Purpose of the Study:
- To develop and validate a selective and quantitative surface plasmon resonance (SPR)-based immunoassay for deoxynivalenol determination.
- To compare the efficacy of three different monoclonal antibodies for DON detection.
Main Methods:
- Preparation and immobilization of a deoxynivalenol-casein conjugate on an SPR sensor chip.
- Development of a competitive inhibition assay using anti-deoxynivalenol monoclonal antibodies.
- Regeneration of the sensor surface with guanidine chloride for reusability.
- Cross-reactivity testing with other trichothecene mycotoxins.
Main Results:
- The SPR immunoassay demonstrated high reusability (>500 cycles) with minimal activity loss.
- The assay showed optimal detection range for deoxynivalenol between 2.5 and 30 ng/mL.
- Minimal cross-reactivity was observed with other tested trichothecene mycotoxins.
- Results from naturally contaminated wheat samples showed good agreement with liquid chromatography/tandem mass spectrometry.
Conclusions:
- The developed SPR immunoassay offers a sensitive, selective, and reusable method for deoxynivalenol quantification.
- Simple sample preparation (acetonitrile extraction and dilution) is sufficient for the assay.
- This method provides a reliable alternative for monitoring deoxynivalenol contamination in food and feed matrices.