DEVELOPMENT OF A MONOCLONAL ANTIBODY-BASED ELISA FOR THE ANTHELMINTIC HYGROMYCIN-B

Monoclonal antibodies were prepared against hygromycin B, a Food and Drug Administration (FDA) approved feed additive used for the control of worms in swine and poultry. Splenocytes from mice immunized with a hygromycin B-ovalbumin conjugate were fused with SP2/0 myeloma cells, and 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 hygromycin B and does not cross-react with structurally similar aminoglycoside antibiotics commonly added to medicated feed. When tissue samples were spiked with hygromycin B, the average percent recoveries at 1, 2, and 4 ppm were 83, 82, and 91%, respectively. The ELISA described here can be used to rapidly screen porcine kidney samples for the presence of hygromycin B residues.