Obtaining a Monoclonal Antibody against a Novel Prometryn-Like Hapten and Characterization of Its Selectivity for Triazine Herbicides

In this study, a previously unreported 3-((4-(isopropylamino)-6-(methylthio)-1,3,5-triazin-2-yl) amino) butyric acid hapten was designed and synthesized. This maximized the exposure of the antigen-determinant isopropyl of prometryn to the immune system in animals to induce the production of anticipated highly specific anti-prometryn antibodies. The hapten has a molecular weight of 285.37 Da. The compound was confirmed by nuclear magnetic resonance hydrogen spectroscopy (H-1 NMR), nuclear magnetic resonance carbon spectroscopy (C-13 NMR), and high-resolution mass spectrometry (HRMS). By using the active ester approach, immunogens and coated antigens were created. Bovine serum albumin (BSA) was used as an immunogen, along with the successfully produced hapten, to immunize mice. The IC50 value of mouse monoclonal anti-prometryn antibody (mAb) 7D4 (the quantity of analyte that generated 50% prometryn inhibition) was 3.9 ng/mL. The anti-prometryn mAb was of the IgG1 subclass. The IC20 (80% binding level (B/B-0) of prometryn)-IC80 (20% binding level (B/B-0) of prometryn) range of the anti-prometryn monoclonal antibody standard curve working range was 0.9-18.1 ng/mL. The prepared mAb has good characteristics because it can specifically recognize prometryn, and the cross-reaction rates for ametryn, desmetryn, and terbumeton were 34.77%, 18.09%, and 7.64%, respectively. The cross-reaction rate with the other seven triazines was less than 1%. The hapten structure proposed can serve as an additional tool for modulating selectivity in detecting triazines.

Automated summary

A novel 3-((4-(isopropylamino)-6-(methylthio)-1,3,5-triazin-2-yl) amino) butyric acid hapten was designed and synthesized to maximize antigen exposure. The hapten was confirmed by NMR and HRMS. Immunogens and coated antigens were created using the active ester approach. Mice were immunized using bovine serum albumin as an immunogen and the hapten. A highly specific monoclonal antibody against prometryn was produced with good characteristics.