Two binding epitopes modulating specificity of immunoassay for fl-agonist detection: Quantitative structure-activity relationship

A growing number of fl-agonists are illegally using for reducing animal fat deposition in animals, but the development of analytical methods always lags behind the emergence of new illegal compounds. Therefore, class specificity antibody-based immunoassays that can detect a great many fl-agonists are important for timely supervision. In this study, a competitive inhibition enzyme-linked immunosorbent assay (ciELISA) based on a clenbuterol monoclonal antibody was developed to recognize 23 fl-agonists and analogues. Holographic and three-dimensional quantitative structure-activity relationship (HQSAR and 3D QSAR) revealed that there are two critical binding epitopes on fl-agonist hapten affecting antibody specificity, and these epitopes have been further validated using a ractopamine antibody with narrow specificity. Tert-butyl at C-2 ' epitope is needed to generate class specific antibodies, and different characteristics of substituents at benzene ring epitope would adjust antibody specificity. This investigation could provide reference for future design of fl-agonist haptens.

Automated summary

This study developed a class-specific antibody immunoassay for detecting various fl-agonists and revealed critical binding epitopes on the fl-agonist hapten affecting antibody specificity through holographic and three-dimensional quantitative structure-activity relationship. The characteristics of substituents at different epitopes adjust antibody specificity.