Chemiluminescence enhancement of enzyme-activated decomposition

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or...

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252700, 424 71, 435 5, 435 6, 435 71, 435 74, 435 791, 435 11, 435 14, 435 79, 435960, 436 71, 436 95, 436172, 436537, 436800, 436805, 436827, C12Q 100

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051457725

ABSTRACT:
Water soluble naturally-occurring and synthetic enhancer substances, generally macromolecular in nature, for example globular proteins that include hydrophobic regions such as bovine serum albumin, and polymeric quaternary ammonium salts such as poly(vinylbenzyltrimethylammonium chloride), which have the ability to inhibit light-emitting fluorophores resulting from the decomposition of chemiluminescent compounds from releasing energy through non-light emitting pathways, are disclosed as permitting the stabilization, and hence increasing the light intensity, of such light-emitting fluorophores in aqueous media as compared to the intensity of the light emitted by the same quantities of such fluorophores in aqueous media in the absence of such enhancer substances. Any chemiluminescent enzymatically cleavable 1,2-dioxetane, for example 3-(2'-spiroadamantane)-4-methoxy-(3"-phosphoryloxy)phenyl-1,2-dioxetane disodium salt, can be used. Auxiliary fluorophores, for example fluorescein and derivatized fluoresceins, that accept energy from fluorophores produced by decomposition of a chemiluminescent compound and in turn emit detectable energy, can also be present. Such enhancer substance/chemiluminescent compound compositions are useful in detecting the presence or determining the concentration of chemical or biological substances in immunoassays, chemical assays and nucleic acid probe assays, and in chemical/physical probe procedures for studying the microstructures of macromolecules.

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