Detection of chemicals by a reporter immunoassay: application to fluoride.

This report describes a concept in which an immunoassay is used indirectly to quantify a nonantigenic very low molecular weight compound participating in a chemical reaction with a haptenic reporter. The detection limit of each reagent is, therefore, governed only by the affinity of the antibodies toward the reporter. Fluoride was used as a model, and silylated estradiol was used as a reporter.

Recent developments for SPIE-IA, a new sandwich immunoassay format for very small molecules.

Recent publications describing new elegant approaches to assay small analytes using noncompetitive format were briefly reviewed. Among these methods, we have developed a new protocol, named SPIE-IA, which involves a cross-linking step achieved using chemical hombifunctional reagents, UV irradiation or free radicals. This new method proved to be useful to detect naturally occurring analyte/antibody complexes or to protect the analytes against degradation by peptidases.

The use of enzyme immunoassays for the detection of abzymatic activities. Application to an enantioselective thioacetal hydrolysis activity.

Relying on the particularly high specificity displayed by antibodies, enzyme immunoassays have proved to be one of the most efficient tools for early detection of the catalytic activities displayed by antibodies. We took advantage of such an assay, namely the Cat-enzyme-linked immunoassay (EIA) approach developed in our laboratories, both to exhibit and characterise an antibody-catalysed thioacetal hydrolysis. Monoclonal antibody (mAb) H3-32 was thus identified to accelerate the hydrolysis reaction of thioacetal substrate (NC9) to vanillylmandelic acid (VMA), with a k(cat) of 0.