FIIA, is based on the introduction of the sample into a carrier stream, which enters the reaction chamber where the immunoreaction takes place. In general in FIIA, the antibodies are immobilized to form an affinity column and analyte is pumped over the column. The loading of the antibodies with analyte is followed by pumping over the column enzyme tracers that compete with the analyte for the limited binding sites of the antibodies. Generally, the indirect format produces a result inversely proportional to the analyte concentration. FIIA can be used with electrochemical, spectrophotometric, fluorimetric and chemilumines-cence detection methods. Conventional UV visible spectrophotometry is also suitable for the FIIA detection of bioligand interactions. FIIA has been used for the detection of diuron and atrazine in water. This method was developed as a cost-effective screen for determining compliance with the European drinking water directive.
FIIA has been successfully used for the detection of pesticides (e.g., triazines [85,86]). At present, FIIA is integrated into different immunosensors.
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