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United States Department of Agriculture

Agricultural Research Service

Title: Hapten Synthesis, Conjugation, and the Production of Monoclonal Antibodies to the Macrolide Antibiotic Tilmicosin

item Beier, Ross

Submitted to: International Conference on Agri-Food Antibodies
Publication Type: Abstract Only
Publication Acceptance Date: October 5, 2001
Publication Date: N/A

Technical Abstract: Tilmicosin is a semi-synthetic macrolide antibiotic that is approved for veterinary use in cattle and swine to combat respiratory disease in the U.S., and is used in Canada for the treatment of pneumonia in lambs. Bovine respiratory disease associated with Mycoplasma bovis and Pasteurella haemolytica serotype 1 can be controlled by a subcutaneous injection of tilmicosin. It is known that most antimicrobial agents have limited ability for cellular penetration. However, tilmicosin concentrates in bovine lung tissue. It would be advantageous to produce a quick and economical competitive enzyme-linked immunosorbent assay (cELISA) method for determining tilmicosin residues in feeds and tissues. It also would be advantageous to produce monoclonal antibodies (Mabs) to tilmicosin that may be used to locate and determine tilmicosin-binding sites to help understand cellular interactions. Molecular modeling was used to help determine an appropriate conjugation site on tilmicosin. A derivative of tilmicosin was used to produce keyhole limpet hemocyanin (KLH) and bovin serum albumin (BSA) conjugates. A novel KLH conjugate was used to immunize BALB/c mice. A BSA amide linkage was produced using 1-ethyl-3-(3-dimethyl- aminopropyl)carbodiimide (EDC), and was used for coating plates. Hybridoma cell lines were generated and those lines that produced antibodies against tilmicosin were isolated. Preliminary studies show that one of the Mabs produced a 40% inhibition at 400 pg/well tilmicosin. Competition studies with tilmicosin, tylosin and other structurally related compounds are presented.

Last Modified: 4/21/2015
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