Alkaline phosphatase, chemiluminescent bioassays

Dioxetanes, labeled with triggers sensitive to the alkaline-phosphatase enzyme, serve as highly sensitive chemiluminescent probes in numerous bioassays. Current applications include immunoassays, membrane-based detection of proteins and nucleic acids, and microplate-based and array-based nucleic-acid detection. ... 

CIEEL is of particular interest for the development of modern chemiluminescent bioassays. The most popular clinical bioassays utilize thermally persistent spiro-adamantyl-substituted dioxetanes with a protected phenolate moiety. These designed 1,2-dioxetanes include an energy source, a fluorophore, and a trigger grouping, and are therefore structurally similar to bioluminescent substrates such as firefly luciferin. Three main commercial dioxetanes 75 are available as one-reagent assays for alkaline phosphatase and are sold under the name of AMPPD (R1 = R2 = H), CSPD (R1 = Cl, R2 = H), and CDP-Star (R1 = R2 = Cl) <2006S1781, 2003ANA279>. These substrates are sensitive to 10 21 mol of alkaline phosphatase in solution. 

The naphtol derivative, Table 13-6, entry 9, is dephosphorylated by an alkaline phosphatase. The resulting naphtol decomposes with chemiluminescent emission and can be used in bioassays to generate a chemiluminescence signal proportional to the concentration of an alkaline phosphatase label. 

A variety of detection schemes have been used with chromatographic immunoassays. The detection can be based on chemical tags that act as chromophores, fluOTO-phores, chemiluminescent agents, or electrochemically active species. Enzymes like (3-galactosidase, alkaline phosphatase, and horseradish peroxidase have also been employed as labels to generate products for detection by absorbance, fluorescence, chemiluminescence, electrochemical, or thermometric measurements. Liposomes, radiolabels, and bioassays have also been used for detection in chromatographic immunoassays. " ... 

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