1. Academic Validation
  2. Alternative Hapten Design for Zearalenone Immunoreagent Generation

Alternative Hapten Design for Zearalenone Immunoreagent Generation

  • Toxins (Basel). 2022 Mar 2;14(3):185. doi: 10.3390/toxins14030185.
Antonio Abad-Fuentes 1 Consuelo Agulló 2 Daniel López-Puertollano 2 Ismael Navarro-Fuertes 2 Antonio Abad-Somovilla 2 Josep Vicent Mercader 1
Affiliations

Affiliations

  • 1 Instituto de Agroquímica y Tecnología de Alimentos, Spanish Council for Scientific Research (CSIC), 28006 Madrid, Spain.
  • 2 Department of Organic Chemistry, University of Valencia, 46010 València, Spain.
Abstract

Appropriate Hapten design and synthesis have been identified as critical steps to generate high-performance immunoreagents and to develop sensitive and selective immunoanalytical methods. Antibodies and immunoassays for the major mycotoxin zearalenone have been reported and marketed. However, zearalenone haptens have mostly been prepared by the oxime active ester technique, and Hapten characterization has generally been poor or non-existent. In the present study, novel haptens of zearalenone with longer linkers and with alternative tethering sites have been designed for immunizing and assay conjugate preparation. All of these molecules were purified and spectroscopically verified, and a structure-activity relationship evaluation was carried out. This approach revealed that the Hapten with the linker at the carbonyl group generated Antibodies with a higher affinity than the Hapten functionalized at the phenyl moiety. Antibodies produced with the latter Hapten, on the other hand, showed lower cross-reactivity values to the major zearalenone metabolites. Finally, similar immunoassay sensitivity was achieved with all of the Antibodies when heterologous haptens were employed. Furthermore, by altering the structure of the competing antigen, the immunoassay selectivity was modified. These results demonstrate that immunochemical methods for zearalenone rapid analysis can still be improved in terms of sensitivity and selectivity.

Keywords

antibody affinity; antibody specificity; hapten design; immunoassay; mycotoxin; spacer arm.

Figures
Products