From the firefly bioluminescence to the dioxetane-based (AMPPD) chemiluminescence immunoassay: A retroanalysis

Chemiluminescent dioxetanes, which are enzymically triggerable, have proved to be a valuable tool in diagnostic clinical applications, In this context, the phenomenon of firefly bioluminescence (the most efficient light-producing system so far) has served as an excellent example to develop new chemiluminescent probes for bioanalysis,;in particular chemiluminescence immunoassays, From the recognition of the molecular details of the firefly bioluminescence, a chemical equivalent has been conceived which possesses the essential features for efficient generation of light, i.e., thermal persistence through spiroadamantyl substitution, spontaneous chemically initiated electron exchange luminescence (CIEEL)-active decomposition after enzymic triggering, efficient light emission as a lasting glow and convenient synthesis by-photooxygenation. This has led to a rationally designed dioxetane, namely the 3-(2'-spiroadamantyl)-4-methoxy-4-(3 ''-phosphoryloxy)-phenyl-1,2-dioxetane (AMPPD) derivative, which serves as a substrate for alkaline phosphatase and, in terms of sensitivity, has surpassed the hazardous radioactive immunoassay probes, This paper-gives a historical account in the form of a retroanalysis of the success story behind the rational design of the first enzymically triggerable, CIEEL-active AMPPD chemiluminescent probe.