Unveil the molecular recognition mechanisms of triazine haptens and monoclonal antibodies: Establishing ic-ELISA methods for triazine herbicide analysis in tea

Abstract

This study introduced two triazine haptens (with 0 or 1 carbon connecting arms at nonisopropyl positions) and developed two heterologous indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) methods. The IC₅₀ values for the 17 triazine pesticides ranged from 0.84 ng/mL to 225.63 ng/mL, and the established method was applied to determine their concentrations in tea samples. This study aimed to address the current debate on the recognition mechanism of triazine haptens with shorter connecting arms. This study produced two new findings by investigating the recognition mechanism of triazine haptens and antibodies. On the one hand, the specific amino acid isoleucine (ILE-219) is crucial for isopropyl recognition. On the other hand, the selection of the connecting site of the triazine hapten has a significant effect on the specificity of the antibody, with the influence of the length of the carbon connecting arm being secondary. Our newly designed haptens show promise for diverse triazine pesticide immunoassay applications. This discovery established a foundational framework for constructing a tailored antibody library for pollutants.