Fluoroimmuno-like assays

The experiment consists in the incubation of the labeled antigen (1) with increasing concentrations of MIP or NIP/CP, and (2) with the same set of concentrations but in the presence of a constant concentration of analyte sufficient to displace significantly the probe from the polymer [22]. Each point in the dilution curve should be determined at least by duplicate and preferably between five and ten replicates so that we can obtain not only the optimum polymer concentration for the assay but also an estimate of the standard deviation of the response at each point along the curves. The curve representing the difference (% bound label) of the bound label in the absence and in the presence of the analyte will render the optimum MIP concentration to be used in the assay. Using this approach we have optimized the amount of polymer required for the development of a fluoroimmuno-like assay for penicillin analysis [36],... 

Fluorometry is a superior optical technique in terms of sensitivity and specificity. Merits of fluoroimmunoassays (FIAs) and fluoroimmuno-like assays (FILAs) include the stability and freedom from hazards of fluorescent labels compared to radioactive tracers, the moderate cost of analysis, the wide availability of the equipment needed, and the potential high sensitivity. In general, the sensitivity of fluorescence measurements is 10- to 1,000-fold higher than the absorption counterparts. 

Fluoroimmuno-Like Assays Based on Direct Measurements... 

Table 5 Analytical characteristics of fluoroimmuno-like assays... 

When the label and the analyte are not identical, as happens in the case of competitive fluoroimmuno-like or enzyme-immunolike assays, we have found it useful to follow the experimental approach suggested by Ekins [22] for the optimization of the antibody concentration in competitive immunoassays. 

Table 6 IC50 values and cross-reactivities (CR) in competitive fluoroimmuno-like MIP assays and...

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