M. Vass, L. Kotkova, I. Diblíková
Feb 11, 2018
Citations
1
Influential Citations
9
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Quality indicators
Journal
Veterinarni Medicina
Abstract
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 immu - nising hapten for the production of monoclonal antibodies against NP AOZ. Monoclonal antibodies were pro - duced 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 produc- tion 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.