Submitted to: Journal of Agricultural and Food Chemistry
Publication Type: Peer reviewed journal
Publication Acceptance Date: 9/1/2005
Publication Date: 12/15/2005
Citation: Beier, R.C., Creemer, L.C., Ziprin, R.L., Nisbet, D.J. 2005. Production and characterization of monoclonal antibodies against the antibotic tilmicosin. Journal of Agricultural and Food Chemistry. 53:9679-9688. Interpretive Summary: Tilmicosin is a drug that is used in cattle, swine, poultry and sheep to combat respiratory disease. We have developed antibodies that recognize tilmicosin, but do not recognize the related drug tylosin. Antibodies are substances that are produced by the immune system in response to foreign substances which enter the body. Once the antibody to a foreign substance is isolated, it can be used in a method to detect the presence of that foreign substance. The antibodies that we isolated to detect tilmicosin may be used in an easy-to-use test to determine levels of tilmicosin in animal tissues, and can be used to localize the site of tilmicosin binding within tissues. These tests would ensure that the levels of tilmicosin were appropriate for treatment of respiratory diseases. Using these antibodies to localize tilmicosin binding in tissues would advance the basic understanding of the action of this drug.
Technical Abstract: Monoclonal antibodies (Mabs) were developed that specifically bind tilmicosin. A KLH and BSA conjugate were used for the immunogen and plate coating antigen, respectively. The conjugates were synthesized using different methods resulting in different linkages. Six hybridoma cell lines were isolated that produced Mabs that competed with tilmicosin, and have IgG1 isotype. The Til-1 and Til-5 Mabs had IC50 values for tilmicosin of 9.6 and 6.4 ng/well, respectively, and a limit of detection at IC20 of 1.84 and 0.89 ng/well, respectively. The Mabs demonstrated high cross-reactivity to the macrolides containing 3,5-dimethyl-piperidine at C20 and the amino sugar at C5. No cross-reactivity was observed for tylosin and other macrolides that did not contain 3,5-dimethylpiperidine. A competitive ELISA was developed for the antibiotic tilmicosin using the developed Mabs. These Mabs may be excellent candidates for the determination and immunolocalization of tilmicosin.