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Application of a new anti-zearalenone monoclonal antibody in different immunoassay formats

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Abstract

Monoclonal antibodies against zearalenone (ZEA) were raised in mice according to the hybridoma technology and applied in different immunochemical techniques. More specifically, three formats based on the competitive direct enzyme immunoassay principle were developed: an enzyme-linked immunosorbent assay (ELISA), a flow-through gel-based immunoassay column and a flow-through membrane-based immunoassay. In ELISA, the 50% inhibitory concentration (IC50) was 0.8 ng/mL, and the limit of detection for ZEA standard solutions was 0.1 ng/mL. The antibodies showed a high ZEA (100%) and α-zearalenol (α-ZOL) (69%) recognition, while cross-reactivities with α-zearalanol, zearalanone, β-zearalenol and β- zearalanol were 42%, 22%, <1% and <1%, respectively. For standard solutions, a cut-off level at 10 ng/mL could be established for the gel- and membrane-based enzyme immunoassays. Assay time of both non-instrumental tests was 25 min for 10 samples. By including a simple sample extraction procedure, the methods were applied to wheat with IC50s in ELISA of 80 and 120 μg/kg (dilution up to 5% and 15% (v/v) of wheat matrix, respectively). The cut-off level of the gel- and membrane-based immunoassays was established at 100 µg/kg. Potentials and limitations of the developed methods were compared. The possible application for multi-mycotoxin analysis of the ELISA method based on a single monoclonal antibody was investigated. Therefore, principal component analysis and partial least squares regression data modelling were used to separate the immunoassay responses of two cross-reactants (ZEA and α-ZOL).

Monoclonal antibodies against zearalenone were raised in mice and applied in three formats based on the competitive direct enzyme immunoassay principle: a microtiter plate enzyme-linked immunosorbent assay (ELISA), a flow-through gel-based immunoassay column and a flow-through membrane-based immunoassay

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Acknowledgements

N.А. Burmistrova benefits from a fellowship granted by the Belgian Federal Science Policy Office in order to promote the S&T cooperation with Central and Eastern Europe. This work was further financially supported by IOF, Ghent University (IOF06/VAL/003).

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Authors and Affiliations

  1. Faculty of Pharmaceutical Sciences, Laboratory of Food Analysis, Ghent University, Harelbekestraat 72, 9000, Ghent, Belgium

    Natalia A. Burmistrova, Carlos Van Peteghem & Sarah De Saeger

  2. Chemistry Faculty, Department of Common and Inorganic Chemistry, Saratov State University, Astrakhanskaya 83, 410012, Saratov, Russia

    Natalia A. Burmistrova, Irina Yu. Goryacheva & Evgenia Yu. Basova

  3. Faculty of Pharmaceutical Sciences, Laboratory of Pharmaceutical Biotechnology, Ghent University, Harelbekestraat 72, 9000, Ghent, Belgium

    Ann-Sophie Franki & Dieter Deforce

  4. Faculty of Medicine and Health Sciences, Laboratory for Molecular Immunology and Inflammation, Ghent University, De Pintelaan 185, 9000, Ghent, Belgium

    Dirk Elewaut & Katrien Van Beneden

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  1. Natalia A. Burmistrova
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  2. Irina Yu. Goryacheva
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  3. Evgenia Yu. Basova
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  4. Ann-Sophie Franki
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  7. Dieter Deforce
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  8. Carlos Van Peteghem
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  9. Sarah De Saeger
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Correspondence to Natalia A. Burmistrova.

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Burmistrova, N.A., Goryacheva, I.Y., Basova, E.Y. et al. Application of a new anti-zearalenone monoclonal antibody in different immunoassay formats. Anal Bioanal Chem 395, 1301–1307 (2009). https://doi.org/10.1007/s00216-009-2913-7

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  • DOI: https://doi.org/10.1007/s00216-009-2913-7

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