Concise and modular synthesis of regioisomeric haptens for the production of high-affinity and stereoselective antibodies to the strobilurin azoxystrobin

The immune response to regioisomeric haptens of azoxystrobin with varied derivatization sites was studied. Based on the Sonogashira and Suzuki–Miyaura couplings and following a straightforward modular design, we have synthesized four haptens with the same linker anchored through C–C bonds and located at different sites of the molecule. The most stereoselective antibodies were produced from immunogens with the spacer arm at a distal position from the β-methoxyacrylate moiety characteristic of strobilurins. Moreover, we observed that assay cross-reactivity was reliant on the functionalization site of the competitor derivative. Finally, the antibody binding site was explored using synthetic chemical analogues.

Graphical abstractFunctionalized regioisomeric haptens of azoxystrobin have been prepared following a modular synthetic strategy, and high-affinity and stereoselective antibodies have been generated thereof.Figure optionsDownload full-size imageDownload as PowerPoint slide