FROM THE FIREFLY BIOLUMINESCENCE TO THE DIOXETANE-BASED (AMPPD) CHEMILUMINESCENCE IMMUNOASSAY - A RETROANALYSIS

Chemiluminescent dioxetanes, which are enzymically triggerable, have p roved to be a valuable tool in diagnostic clinical applications, In th is context, the phenomenon of firefly bioluminescence (the most effici ent 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 genera tion of light, i.e., thermal persistence through spiroadamantyl substi tution, spontaneous chemically initiated electron exchange luminescenc e (CIEEL)-active decomposition after enzymic triggering, efficient lig ht emission as a lasting glow and convenient synthesis by-photooxygena tion. This has led to a rationally designed dioxetane, namely the 3-(2 '-spiroadamantyl)-4-methoxy-4-(3 ''-phosphoryloxy)-phenyl-1,2-dioxetan e (AMPPD) derivative, which serves as a substrate for alkaline phospha tase and, in terms of sensitivity, has surpassed the hazardous radioac tive immunoassay probes, This paper-gives a historical account in the form of a retroanalysis of the success story behind the rational desig n of the first enzymically triggerable, CIEEL-active AMPPD chemilumine scent probe.