DEVELOPMENT OF A MONOCLONAL ANTIBODY-BASED ELISA FOR THE ANTHELMINTICHYGROMYCIN-B

Monoclonal antibodies were prepared against hygromycin B, a Food and D rug Administration (FDA) approved feed additive used for the control o f worms in swine and poultry. Splenocytes from mice immunized with a h ygromycin B-ovalbumin conjugate were fused with SP2/0 myeloma cells, a nd hybridomas secreting antibodies against hygromycin B were selected and cloned. An assay using the antibody with the highest affinity for hygromycin B is described. The monoclonal antibody is specific for hyg romycin B and does not cross-react with structurally similar aminoglyc oside antibiotics commonly added to medicated feed. When tissue sample s were spiked with hygromycin B, the average percent recoveries at 1, 2, and 4 ppm were 83, 82, and 98%, respectively. The ELISA described h ere can be used to rapidly screen porcine kidney samples for the prese nce of hygromycin B residues.