US4246340A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Method and reagent for bioluminiscence
Assignee: Pfizer Health AB
•
Filing: 1979-05-30
•
Granted: 1981-01-20
•
Expires: 1999-05-30
Abstract
Abstract An improvement in the process for the determination of the concentration of adenosinetriphosphate (ATP) in ATP-converting reactions wherein the concentration is a function of the intensity of light emitted when a sample to be assayed is contacted with a bioluminescent reagent based on D-luciferin, luciferase and magnesium ions or certain other metal ions, the emitted light resulting from ATP and D-luciferin becoming bound to the luciferase, comprises the use of one or more competitive inhibitors such as a D-luciferin analogue, or analogues.
Inventors
- 👤 Arne T. Lundin
- 👤 Arne Myhrman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 00:48
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/66
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
82 words
A process for the determination of ATP concentrations by contacting the sample to be assayed, with a bioluminescence reagent based on D-luciferin, luciferase and metal ions, a reaction being obtained wherein ATP and D-luciferin are bound to the luciferase and light is emitted, and by measuring the intensity of the emitted light, which intensity is a measure of the ATP concentration, characterized by using in the determination at least one competitive inhibitor of the reaction in the form of a D-luciferin analogue.
Claim 2
Dependent Claim
82 words
A process for the determination of ATP concentrations by contacting the sample to be assayed, with a bioluminescence reagent based on D-luciferin, luciferase and metal ions, a reaction being obtained wherein ATP and D-luciferin are bound to the luciferase and light is emitted, and by measuring the intensity of the emitted light, which intensity is a measure of the ATP concentration, characterized by using in the determination at least one competitive inhibitor of the reaction in the form of a D-luciferin analogue.
Claim 3
Dependent Claim
29 words
A process according to claim 1, characterized by using the D-luciferin analogue at a concentration which gives an inhibition of at least 25%, preferably 50-90%, of the light intensity.
Claim 4
Dependent Claim
16 words
A process according to claim 1 or 2, characterized by using L-luciferin as the D-luciferin analogue.
Claim 5
Dependent Claim
16 words
A process according to claim 1, characterized in that also pyrophosphate is used in the determination.
Claim 6
Dependent Claim
25 words
A process according to claim 4, characterized by using the pyrophosphate at a highest concentration of 10 -4 M, preferably at most 10 -5 M.
Claim 7
Dependent Claim
17 words
A process according to claim 1, characterized in that also coenzyme A is used in the determination.
Claim 8
Dependent Claim
20 words
A process according to claim 1, characterized by using a reagent containing luciferase that has been purified by isoelectric focusation.
Claim 9
Dependent Claim
24 words
A process according to claim 1, characterized by using a reagent containing luciferase with an additive of bovine serum albumine, thiol compounds and/or EDTA.
Claim 10
Dependent Claim
38 words
A bioluminescence reagent for use in the determination of ATP-concentrations, which reagent is based on D-luciferin, luciferase and metal ions, characterized in that it also contains at least one competitive inhibitor in the form of a D-luciferin analogue.
Claim 11
Dependent Claim
38 words
A bioluminescence reagent for use in the determination of ATP-concentrations, which reagent is based on D-luciferin, luciferase and metal ions, characterized in that it also contains at least one competitive inhibitor in the form of a D-luciferin analogue.
Claim 12
Dependent Claim
31 words
A reagent according to claim 9, characterized in that the D-luciferin analogue is present at a concentration which gives an inhibition of at least 25%, preferably 50-90%, of the light intensity.
Claim 13
Dependent Claim
16 words
A reagent according to claim 9 or 10, characterized in that the D-luciferin analogue is L-luciferin.
Claim 14
Dependent Claim
13 words
A reagent according to claim 9, characterized in that it also contains pyrophosphate.
Claim 15
Dependent Claim
27 words
A reagent according to claim 12, characterized in that the pyrophosphate is present at a highest concentration of 10 -4 M, preferably at most 10 -5 M.
Claim 16
Dependent Claim
14 words
A reagent according to claim 9, characterized in that it also contains coenzyme A.
Claim 17
Dependent Claim
19 words
A reagent according to claim 9, characterized in that it contains luciferase that has been purified by isoelectric focusation.
Claim 18
Dependent Claim
23 words
A reagent according to claim 9, characterized in that it contains luciferase with an additive of bovine serum albumine, thiol compounds and/or EDTA.
References Cited (Backward Citations)
US3933592A
Cited
US3616253A
Cited
US3575811A
Cited
US3575812A
Cited
US3929586A
Cited
US4001088A
Cited
US4080265A
Cited
GB2001434A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1984-01-20 | 1984-07-21 | 1985-01-20 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1988-01-20 | 1988-07-21 | 1989-01-20 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1992-01-20 | 1992-07-21 | 1993-01-20 | $7700.00 | Window Closed / Lapsed |