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Production and characterisation of monoclonal antibodies for the detection of AOZ, a tissue bound metabolite of furazolidone

Author

Listed:
  • M. Vass

    (Veterinary Research Institute, Brno, Czech Republic)

  • L. Kotkova

    (Veterinary Research Institute, Brno, Czech Republic)

  • I. Diblikova

    (Veterinary Research Institute, Brno, Czech Republic)

  • Z. Nevorankova

    (Veterinary Research Institute, Brno, Czech Republic)

  • K.M. Cooper

    (Queen's University Belfast, Department of Veterinary Science, Northern Ireland, UK)

  • D.G. Kennedy

    (Chemical Surveillance Branch, Veterinary Sciences Division, Department of Agriculture & Rural Development, Belfast, Northern Ireland, UK)

  • M. Franek

    (Veterinary Research Institute, Brno, Czech Republic)

Abstract

3-amino-2-oxazolidinone (AOZ) is a tissue bound toxic metabolite derived from the nitrofuran antibiotic, furazolidone. AOZ is detected in the derivatised form of 3-{[(2-nitrophenyl) methylene] amino}-2-oxa-zolidinone (NP AOZ). 3-{[(3-carboxyphenyl)-methylene] amino-2-oxazolidinone (CP AOZ) was used as the immunising hapten for the production of monoclonal antibodies against NP AOZ. Monoclonal antibodies were produced using hybridomas from the fusion of murine myeloma cells and spleen cells isolated from BALB/c mice immunised with CP AOZ-ethylenediamine-human serum albumin (CP AOZ-ed-HSA). The antibody production in ascitic fluids from clones 3B8/2B9 and 2D11/A4 was monitored during a 16 month period. Repeated cultures of these hybridomas, followed by injection into mice and cloning did not change the assay parameters. Clone 2D11/A4 exhibited long term stability in antibody production throughout the experiment whereas clone 3B8/2B9 demonstrated variability in particular antibody yields whilst retaining assay sensitivity. Reasons for this production variability in clones are discussed. In an optimised direct ELISA format, the antibodies exhibited a 50% binding inhibition in the range of 0.52-1.15 ng/ml with NP AOZ (0.22 -0.50 ng/ml, respective AOZ equivalents) and showed high specificity towards this analyte. The sensitivity of monoclonal antibodies incorporated into the ELISA is compatible with the European Union MRLP and is currently in use for routine analysis.

Suggested Citation

  • M. Vass & L. Kotkova & I. Diblikova & Z. Nevorankova & K.M. Cooper & D.G. Kennedy & M. Franek, 2005. "Production and characterisation of monoclonal antibodies for the detection of AOZ, a tissue bound metabolite of furazolidone," Veterinární medicína, Czech Academy of Agricultural Sciences, vol. 50(7), pages 300-310.
    • Handle: RePEc:caa:jnlvet:v:50:y:2005:i:7:id:5627-vetmed
    DOI: 10.17221/5627-VETMED
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    Citations

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    Cited by:

    1. M. Franek & I. Diblikova & M. Vass & L. Kotkova & K. Stastny & K. Frgalova & K. Hruska, 2006. "Validation of a monoclonal antibody-based ELISA for the quantification of the furazolidone metabolite (AOZ) in eggs using various sample preparation," Veterinární medicína, Czech Academy of Agricultural Sciences, vol. 51(5), pages 248-257.
    2. M. Vass & K. Hruska & M. Franek, 2008. "Nitrofuran antibiotics: a review on the application, prohibition and residual analysis," Veterinární medicína, Czech Academy of Agricultural Sciences, vol. 53(9), pages 469-500.

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