Amplified assay of alkaline phosphatase using flavinadeninedinucleotide phosphate as substrate

摘要: Abstract A simple to use, robust, quantitative, and extremelysensitive colorimetric assay for alkaline phosphatase (EC 3.1.3.1), designed be used as a detection system in diagnostic assays employing antibodies or gene probes, is described. This technology based on the novel principle of prosthetogenesis, according which purpose-designed substrate (a prosthetogen) primary analyte-linked enzyme label hydrolyzed produce prosthetic group detector system. The prosthetogen employed here derivative FAD phosphorylated at 3′-position ribose ring (FADP), phosphatase, D-amino-acid oxidaselhorseradish peroxidase-coupled Essentially each turnover every molecule produces oxidase detection. Thus enormous amplification initial signal achieved short time periods because relatively high number FADP. can formatted stable, preformed, freeze-dried preparation containing all analytical components, reconstituted simply by addition buffer solution. methodology quantitate less than 0.1 amol 30 min 25°C using microtiter plates.