AI Article Synopsis
- The production of an antibody to detect toltrazuril and its metabolite ponazuril faced challenges due to the difficulty of attaching these substances to a carrier protein.
- Researchers identified trifluoraminoether as a suitable compound to mimic the target structure and successfully conjugated it to two different carrier proteins for immunization in rabbits.
- The resulting polyclonal antibody, named R609, demonstrated high sensitivity and specificity for toltrazuril and its metabolites, marking the first time an antibody has been developed that can accurately bind to these coccidiostats without cross-reacting with similar compounds.
Article Abstract
The production of an antibody to detect toltrazuril or its metabolite ponazuril is complicated due to structural constraints of conjugating these coccidiostats to a carrier protein. Therefore a search was carried out for a compound that shared a common substructure to use as an antigen mimic. The chosen compound, trifluoraminoether, was conjugated to two carrier proteins (HSA and BTG) and used in the immunisation of six rabbits. Two immunogen doses (1 mg and 0.1 mg) were also used. All six rabbits produced an immunological response to the hapten regardless of the carrier protein or immunogen dose used. The most sensitive polyclonal antibody produced, designated R609, was subsequently characterised. This antiserum exhibited an IC50 of 18 ng ml(-1) using a competitive ELISA format. Cross reactivity studies show that this serum is specific for toltrazuril and its metabolites (toltrazuril sulfoxide and toltrazuril sulfone) but does not cross-react with other coccidiostats such as halofuginone, nitroimidazoles or nicarbazin. This is the first reported production of an antibody capable of specifically binding toltrazuril and ponazuril.
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| http://dx.doi.org/10.1039/b301898a | DOI Listing |
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