PRODUCTION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES AGAINST THE POULTRY COCCIDIOSTAT HALOFUGINONE

A succinyl derivative of halofuginone (Hal) was covalently linked to k eyhole limpet hemocyanin to provide an immunogen for production of ant i-Hal antibodies. This resulted in six hybridoma cell lines producing Hal-specific monoclonal antibodies (Mabs) of moderate to high affinity . All 6 Mabs were highly specific for Ha! relative to 12 compounds wit h similar structural components. Fifty percent inhibition of control v alues (IC50) for the Mabs ranged from 0.19 to 12.0 ng of Hal hydrobrom ide/well. One Mab (IC50 = 0.19 ng/well) was used to develop a competit ive enzyme-linked immunosorbent assay (cELISA) for Hal in chicken seru m. Interference due to serum matrix effects was effectively eliminated by I-fold dilution of the sample with assay buffer. The minimum detec table amount of Hal was 0.05 ng/well. Analysis of Hal-fortified serum (6.25-50 ppb) showed excellent agreement (92-100 %) with the amount of Hal added.