Alternative Hapten Design for Zearalenone Immunoreagent Generation

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.

Immunological strip sensor for the rapid determination of niacin in dietary supplements and foods

“…, or by reducing unnecessary chemical groups such as ester bonds, with as little change as possible to the original structure. 39–41 Due to the relatively simple structure of niacin itself, the molecular weight was small. In this experiment, for the chemical structure of niacin, we designed from two sites.…”

“…In general, haptens were designed to be able to couple to carrier proteins by adding chemical groups such as carboxyl groups, amino groups, etc., or by reducing unnecessary chemical groups such as ester bonds, with as little change as possible to the original structure. [39][40][41] Due to the relatively simple structure of niacin itself, the molecular weight was small. In this experiment, for the chemical structure of niacin, we designed from two sites.…”

Improvement of the sensitivity of lateral flow systems for detecting mycotoxins: Up‐to‐date strategies and future perspectives

“…In another research, Abad‐Fuentes et al. (2022) designed novel ZEN haptens with longer linkers and alternative binding sites, demonstrating that haptens with linkers at carbonyl groups produced antibodies that had a higher affinity and exhibited a lower cross‐reactivity. Another determinant factor in monoclonal antibodies’ analytical quality and suitability is the screening method used during their production (Sakaguchi et al., 2021).…”

Development of a colloidal gold strip assay for the detection of total homocysteine in serum samples

“…17 The structure of the hapten is very important for the preparation of suitable antibodies, and in general, the more complex or specially designed a hapten is, the easier it is to obtain antibodies that meet a requirement. 18 Hcy is an amino acid derivative with a very simple structure, and it is difficult to prepare antibodies that directly recognize this molecule. However, when Hcy is hydrolyzed (Hcy is hydrolyzed into SAH under the action of SAH hydrolase 19 ), the concentration of Hcy can be indirectly measured by the detection of † Electronic supplementary information (ESI) available.…”