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US 5,004,565 AGranted
Method and compositions providing enhanced chemiluminescence from 1,2-dioxetanes
AssigneeBoard of Governors of Wayne State University
InventorArthur P. Schaap
Life of this patent
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Abstract
A method and compositions including a 1,2-dioxetane and a fluorescent compound is described. In particular, enzymatic triggering of a triggerable 1,2-dioxetane admixed with a surfactant and the fluorescent compound attached to a hydrocarbon to provide a co-surfactant in a micelle or other structure providing close association of these molecules is described. The method and compositions are useful in immunoassays and in DNA probes used for various purposes.
Claims35
A composition which generates light upon triggering which comprises in admixture:
- (a) a fluorescent compound;
- and
- (b) a stable 1,2-dioxetane of the formula:
- ##STR32## wherein ArOX represents an aryl group substituted with an X-oxy group which forms an unstable oxide intermediate 1,2-dioxetane compound when triggered to remove X by an activating agent so that the unstable 1,2-dioxetane compound decomposes and releases electronic energy to form light and two carbonyl containing compounds of the formula:
- ##STR33## wherein X is a chemically labile group which is removed by the activating agent to form the unstable oxide intermediate 1,2-dioxetane and wherein A are passive organic groups which allow the light to be produced wherein the stable 1,2-dioxetane is decomposed with the activating agent wherein the dioxetane and the fluorescent compound are provided in a closely spaced relationship in the presence of a surfactant, micelle, liposome, reversed micelle, microemulsion, film, including a monolayer, or polymer and wherein the fluorescent compound accepts the electronic energy generated upon decomposition of the unstable oxide intermediate and produces a more intense light than is produced by the triggering of the dioxetane alone.
2 more elements
A composition which generates light upon triggering which comprises in admixture:
- (a) a fluorescent compound;
- and
- (b) a stable 1,2-dioxetane compound of the formula:
- ##STR34## wherein one of R.sub.1 and R.sub.2 together and R.sub.3 and R.sub.4 together can be joined as spirofused alkylene which can contain hetero atoms selected from the group consisting of N, S, O and P in addition to carbon and aryl rings, wherein at least one of R.sub.1 and R.sub.2 or R.sub.3 and R.sub.4 is an aryl group substituted with an X-oxy group which forms an unstable oxide intermediate 1,2 dioxetane compound when triggered to remove X by an activating agent selected from acids, bases, salts, enzymes, inorganic and organic catalysts and electron donors so that the unstable 1,2-dioxetane compound decomposes and releases electronic energy to form light and two carbonyl containing compounds of the formula:
- ##STR35## wherein those of R.sub.1, R.sub.2, R.sub.3 or R.sub.4 which are unsubstituted by an X-oxy group are organic groups which provide stability for the stable 1,2-dioxetane compound and wherein X is a chemically labile group which is removed by the activating agent to form the unstable oxide intermediate wherein the stable 1,2-dioxetane is decomposed with the activating agent, wherein the dioxetane and the fluorescent compound are provided n a closely spaced relationship in the presence of a surfactant, micelle, liposome, reversed micelle, microemulsion, film, including a monolayer, or polymer and wherein the fluorescent compound accepts the electronic energy generated upon decomposition of the unstable oxide intermediate and produces a more intense light than is produced by the triggering of the dioxetane alone.
2 more elements
A composition which generates light upon triggering which comprises in admixture:
- (a) a fluorescent compound;
- and
- (b) a stable dioxetane compound of the formula:
- ##STR36## wherein R.sub.1 is selected from alkyl, alkoxy, aryloxy dialkylamino, trialkyl or aryl silyloxy and aryl groups including spirofused aryl groups with R.sub.2, wherein R.sub.2 si an aryl group which can include R.sub.1 and is substituted with an X-oxy group which forms an unstable oxide intermediate 1,2-dioxetane compound when triggered to remove X by an activating agent selected from acids, bases, salts, enzymes, inorganic and organic catalysts and electron donors so that the unstable 1,2-dioxetane compound decomposes and releases electronic energy to form light and two carbonyl containing compounds of the formula:
- ##STR37## wherein X is a chemically labile group which is removed by the activating agent to form the unstable oxide intermediate and wherein R.sub.3 and R.sub.4 are selected from aryl, heteroalkyl and alkyl groups which can be joined together as spirofused polycyclic alkyl and polycyclic aryl groups wherein the stable 1,2-dioxetane is decomposed with the activating agent, wherein the dioxetane and the fluorescent compound are provided in a closely spaced relationship in the presence of a surfactant, micelle, liposome, reversed micelle, microemulsion, film, including a monolayer, or polymer and wherein the fluorescent compound accepts the electronic energy generated upon decomposition of the unstable oxide intermediate and produces a more intense light than is produced by the triggering of the dioxetane alone.
2 more elements
Dependencies are inferred from the claim text.