Light-activated in vitro assay process for luciferase bioluminescence
OwnerYahia A. Gawadinventor
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Abstract
There is provided a process of inducing luminescent emission from a luciferase bioluminescent reaction particularly useful in binding assays. A luciferase bioluminescent combination, together with an inactive, caged trigger compound such as a cofactor, is subjected to photonic radiation, so as to release the trigger compound in active form, and thereby cause substantially instantaneous reaction of the active trigger compound so released with the luciferase combination, to induce photonic emission which can be detected and measured.
Claims18
A process of inducing luminescent emission from a luciferase bioluminescent reaction mixture which comprises:
- preparing a reaction mixture including components required for a specific luciferase bioluminescent reaction, said components including a luciferase, a luciferin chosen for specific interaction with said luciferase, and at least one cofactor therefor, one said component being initially included in the reaction mixture in an inactive, caged form;
- subjecting the reaction mixture to photonic radiation of appropriately chosen characteristics to cause release of the one said component from its caged form, in active form thereby triggering the bioluminescent reaction;
- and detecting the photonic emission so caused.
The process of any preceding claim wherein the luciferase is firefly luciferase, bacterial luciferase or fungal luciferase.
The process of any preceding claim wherein the cofactor is selected from adenosine triphosphate (ATP), nicotinamide-adenine dinucleotide phosphate (NADP), nicotinamide-adenine dinucleotide (NAD) and flavin mononucleotide (FMN).
The process of any preceding claim wherein the reaction mixture is utilized to detect the presence of an analyte in a biological fluid by binding reactions.
A process of inducing luminescent emission from a luciferase bioluminescent reaction, which comprises subjecting a luciferase/luciferase emitter, in the presence of inactive, caged, appropriately chosen trigger compound, to photonic radiation adapted to release the trigger compound in active form, and causing substantially instantaneous reaction of the active trigger compound so released with the luciferase-luciferase emitter so as to induce photonic emission therefrom.
A composition of matter including a luciferase/luciferase emitter, and a trigger compound which is a photolabile, caged cofactor adapted to release the cofactor in active form upon appropriate photonic energy input thereto, and being chosen to interact with the luciferase/luciferase emitter combination to initiate bioluminescence when the cofactor is photonically released in its active form.
Dependencies are inferred from the claim text.