Influence of the hapten design on the development of a competitive ELISA for the determination of the antifouling agent Irgarol 1051 at trace levels.

ABSTRACT

Enzyme-linked immunosorbent assays (ELISAs) with a high detectability have been developed for determination of the antifouling agent Irgarol 1051. The features of the resulting assays have been rationalized by using molecular mechanic calculations (MM2+) to correlate the chemical structure of different immunizing haptens and the corresponding avidities of the obtained antisera. The ability of Irgarol 1051 to compete for the antibody binding sites with 11 horseradish peroxidase enzyme tracers, differing in the chemical structures of the hapten, has been investigated. The present paper demonstrates that high-quality antibodies and, therefore, immunoassays reaching very low detection limits could be predicted by molecular modeling studies of the analyte conformations and of the immunizing haptens' geometries, hydrogen-bonding capabilities, and electronic distributions. Two of the ELISAs obtained have been optimized to obtain reproducible immunoassays. The dynamic ranges of both assays are between 30 and 200 ng/L, and the limits of detection are approximately 16 ng/L. The reported immunoassays have been evaluated and validated by analyzing spiked and real seawater samples. Irgarol 1051 has been found to be present in two of the geographical locations analyzed at concentration levels dependent on the time of year. The analytical results obtained with these immunoassays have been validated by chromatographic methods.