Details

Autor(en) / Beteiligte

• Shelver, W.L
• Kim, H.J
• Li, Q.X

Titel

Development of a monoclonal antibody-based enzyme-linked immunosorbent assay for the β-adrenergic agonist zilpaterol

Ist Teil von

• Journal of agricultural and food chemistry, 2005, Vol.53 (9), p.3273-3280

Erscheinungsjahr

2005

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• Zilpaterol is a beta-adrenergic agonist approved for use as a growth promoter in cattle in South Africa and Mexico but not in the European Union, United States, or Asia. Here, we report the development of a monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) for zilpaterol. Mice immunized with zilpaterol-butyrate-keyhole limpet hemocyanin were utilized for monoclonal antibody generation whereas zilpaterol-butyrate-bovine serum albumin was used as a coating antigen for ELISA. Thirteen clones were isolated, and after the initial sensitivity and isotyping experiments, three clones were selected for further ELISA optimization. Studies indicated that the optimum pH was near 7.4. Clone 3H5 had the highest sensitivity to zilpaterol and some interaction with clenbuterol and terbutaline at high concentrations but not other N-alkyl bamethane, (-)-isoproterenol, (+)-isoproterenol, metaproterenol, or salbutamol or N-arylalkyl (fenoterol, isoxsuprine, ractopamine, or salmeterol) beta-agonists tested. However, clone 3H5 was not functional at high salt concentrations, which precluded further development for urine analysis. Clone 2E10 showed increased sensitivity as salt concentrations were increased and did not cross-react with any of the structural analogues tested. However, its sensitivity to salt and urine concentration changes could cause high variability. Clone 7A8 showed good sensitivity and only a modest change with the salt concentration changes. Clone 7A8 also demonstrated smaller changes in IC50 and B0 with increasing sheep urine or cattle urine concentrations as compared to clones 2E10 or 3H5 and, thus, was selected for further development. The IC50 for all of the antibodies showed exponential increases with increasing organic solvents concentrations, making it desirable to minimize solvent levels. In conclusion, a sensitive, specific zilpaterol monoclonal antibody-based ELISA has been developed that can serve as a rapid screening assay.