Quenching reagents and assays for enzyme-mediated luminescence
First Claim
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1. A method of assaying enzyme-mediated luminescence reactions comprising:
- (a) initiating at least one enzyme-mediated luminescence reaction;
then(b) quantifying luminescence energy produced by the luminescence reaction; and
then(c) quenching photon emission from the enzyme-mediated luminescence reaction by introducing at least one quench reagent to the luminescence reaction.
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Abstract
The present invention relates to single and dual-reporter luminescence assays utilizing general and specific reagents to quench enzyme-mediated reactions. In one embodiment of the invention, a reagent is added to the assay which non-specifically quenches enzyme-mediated luminescent reactions. In another embodiment of the invention, a reagent is added to the assay which simultaneously quenches one enzyme-mediated luminescent reaction while activating another distinct enzyme-mediated luminescent reaction. An assay kit containing specific quench reagents, and the reagents themselves are also disclosed.
163 Citations
60 Claims
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1. A method of assaying enzyme-mediated luminescence reactions comprising:
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(a) initiating at least one enzyme-mediated luminescence reaction;
then(b) quantifying luminescence energy produced by the luminescence reaction; and
then(c) quenching photon emission from the enzyme-mediated luminescence reaction by introducing at least one quench reagent to the luminescence reaction. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45)
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46. A method of assaying enzyme-mediated luminescence reactions within multisample assay plates comprising:
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(a) initiating at least one enzyme-mediated luminescence reaction; (b) quantifying luminescence energy produced by the luminescence reaction; and (c) quenching the enzyme-mediated luminescence reaction by introducing at least one quench reagent to the luminescence reaction; whereby refractive interference of light between samples within the multisample assay plate is eliminated. - View Dependent Claims (47)
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48. An enzyme-mediated luminescence reaction assay kit comprising:
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at least one quench reagent composition capable of quenching photon emission from an enzyme-mediated luminescence reaction by a factor of at least 1,000-fold; a suitable first container, said at least one quench reagent composition disposed therein; at least one functional enzyme substrate corresponding to the enzyme-mediated luminescence reaction being assayed; a suitable second container, said at least one functional enzyme substrate disposed therein; and instructions for use. - View Dependent Claims (49, 50, 51)
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52. A dual-reporter enzyme-mediated luminescence reaction assay kit comprising:
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a first functional enzyme substrate corresponding to a first enzyme-mediated luminescence reaction being assayed; a suitable first container, said first functional enzyme substrate disposed therein; a quench-and-activate composition comprising quench reagents capable of quenching photon emission from the first enzyme-mediated luminescence reaction by a factor of at least 1,000-fold, and a second and distinct functional enzyme substrate corresponding to a second and distinct enzyme-mediated luminescence reaction; a suitable;
second container, said quench-and-activate composition disposed therein; andinstructions for use. - View Dependent Claims (53, 54, 55, 56)
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57. Enzyme-mediated luminescence reaction quench reagents capable of reducing photon emissions from enzyme-mediated luminescence reactions comprising:
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iodide, iodine, sulfate, nitrate, iso-propanol, 2-(4-aminophenyl)-6-methylbenzothiazole, dimethyldecylphosphine oxide, pyrophosphate, benzothiazole, 2-phenylbenzothiazole, n-butanol, trans-1,2,-diaminocyclohexane-N,N,N'"'"',N'"'"'-tetraacetic acid, 2-(6'"'"'-hydroxy-2'"'"'-benzothiazolyl)-thiazole-4-carboxylic acid, ethylenediaminetetraacetic acid, 2(o-hydroxyphenyl)benzothiazole, India fountain pen ink, adenosine 5'"'"'-triphosphate 2'"'"',3'"'"'-acyclic dialcohol, periodate oxidized, borohydride reduced sodium dodecyl sulfate, citric acid, Tween®
20,Triton®
X-100,and mixtures thereof. - View Dependent Claims (58, 59, 60)
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Specification