Analytical Properties of Immunosensors Working in Organic Media

The compatibility between organic solvents and immunoreagents was studied for the development of immunosensors for pesticides. On the basis on heterogeneous competitive enzyme formats, three assay types were used: immobilized antibodies (direct format), immobilized hapten conjugates (indirect format), and capture format based on immobilized protein A/G. In all cases, peroxidase enzyme label and fluorometric detection were employed. Initial findings were developed in batch working with the immunoreagents in different solvent systems (pure and mixed) to retrieve basic information about their performances. Monoclonal and polyclonal antibodies for carbaryl and 1-naphthol were used to develop sensors in different organic solvent mixtures. Polyclonal antibodies showed better sensitivity than monoclonal ones in comparable conditions. Sensitivity was better in the more polar solvents, methanol being the best. Comparison with an aqueous immunosensor showed lower sensitivity but better selectivity.