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Submitted to: Meeting Abstract
Publication Type: Abstract Only Publication Acceptance Date: 1/1/2004 Publication Date: 5/10/2004 Citation: Shelver, W.L., Smith, D.J. 2004. Development of an enzyme-linked immunosorbent assay for beta-agonist zilpaterol. EuroResidue V, May 10-12, 2004, Noordwijkerhout, The Netherlands. Interpretive Summary: Abstract only. No Interpretive Summary needed. Technical Abstract: Zilpaterol is a beta-adrenergic agonist approved for use in cattle in South Africa and Mexico but not in the EU or USA. Therefore, we developed an ELISA for zilpaterol. Zilpaterol was reacted with ethyl-4-bromobutyrate followed by reflux with 0.1 M potassium hydroxide; the hapten was reacted with bovine serum albumin or keyhole limpet hemocyanin using EDAC. Anti-zilpaterol antibodies were collected from goats. The average IC50 was 3.94 +/- 0.48 ng/mL (n = 25). The antibody did not cross react with N-alkyl [clenbuterol, bamethane, metaproterenol, salmeterol, salbuterol, (-) isoproterenol, and (+) isoproterenol] or di-aromatic (ractopamine, fenoterol, dobutamine, and isoxsuprine) beta-agonists. The zilpaterol ELISA was tolerant of 5% ethanol or methanol; salt (0 - 1M) did not alter the B0 or IC50s. With pH<6 or >8.8 buffer, the IC50s increased relative to ELISAs run at pH 7.4. In summary, a sensitive, specific zilpaterol ELISA has been developed that can serve as a rapid screening assay. |
