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.

Simple analytical model of biosensor competition analysis for detection of low-molecular-weight analytes

“…(5) describes the experimental data well not only in the equilibrium state but also far from equilibrium is interesting [12]. This is probably due to features of the left part of Eq.…”

Disulfide Symmetric Dimers as Stable Pre‐Hapten Forms for Bioconjugation: A Strategy to Prepare Immunoreagents for the Detection of Sulfophenyl Carboxylate Residues in Environmental Samples

“…The possibility of tailor-made bioreceptors has been a matter of investigation mainly by using theoretical chemistry tools or molecular bioengineering methods. Thus, immunochemical methods have often been exploited for their capability to direct antibody specificity through appropriate hapten design by using theoretical methods and models [20,[36][37][38][39] or recombinant antibody methodologies. [40,41] However, as reported previously, addressing antibody recognition towards the common sulfonic group of the SPCs failed to produce generic antibodies.…”

Application of computer-assisted molecular modeling for immunoassay of low molecular weight food contaminants: A review

“…[30] Antibiotics Ceftiofur CAChe software with the MM2 force field was used to calculate the minimum energy conformations of all structures. Molecular orbitals were calculated with the Alvarez collected parameters, the electrostatic potentials were superimposed on the 3D b isosurfaces, and hapten design, CR c and Ab d recognition studies were conducted.…”

“…Many immunoassays used for detection of low molecular weight food contaminants such as an algaecide [30], antibiotics [6,16,[31][32][33][34][35], cork taint [36], herbicides [37][38][39][40], mycotoxins [41,42], pesticides [43][44][45][46][47][48], veterinary drugs [49][50][51] and other chemicals [52][53][54][55][56] have been developed with the use of CAMM methods (Table 1). CAMM can be used to assist in hapten design, study antibody-antigen recognition in cross-reactivity studies and model antibody binding sites (Table 1).…”