Instituto de Agroquímica y Tecnología de Alimentos, Spanish Council for Scientific Research (CSIC), 28006 Madrid, Spain.
Department of Organic Chemistry, University of Valencia, 46010 València, Spain.
Toxins (Basel). 2022 Mar 2;14(3):185. doi: 10.3390/toxins14030185.
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.
合适的半抗原设计和合成被认为是产生高性能免疫试剂和开发敏感、选择性免疫分析方法的关键步骤。已经有报道和市售用于主要霉菌毒素玉米赤霉烯酮的抗体和免疫分析。然而，玉米赤霉烯酮的半抗原大多是通过肟活性酯技术制备的，对半抗原的表征通常很差或不存在。在本研究中，为了免疫和分析偶联物的制备，设计了具有更长连接子和替代连接位点的新型玉米赤霉烯酮半抗原。所有这些分子都经过了纯化和光谱验证，并进行了结构-活性关系评估。这种方法表明，在羰基处具有连接子的半抗原产生的抗体具有比在苯部分官能化的半抗原更高的亲和力。另一方面，用后一种半抗原产生的抗体对主要玉米赤霉烯酮代谢物的交叉反应值较低。最后，当使用异源半抗原时，所有抗体都实现了类似的免疫分析灵敏度。此外，通过改变竞争抗原的结构，可以改变免疫分析的选择性。这些结果表明，玉米赤霉烯酮快速分析的免疫化学方法在灵敏度和选择性方面仍有改进的空间。
