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US4264727A
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Assaying of digoxin by means of bioluminescence and inhibition of NA+-+ -ATPase
Assignee: LUMAC INTERNATIONAL NV A CORP OF NETHERLANDS ANTILLES
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Filing: 1979-07-24
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Granted: 1981-04-28
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Expires: 1999-07-24
Abstract
Abstract A method is described for quantitative measurement of digoxin and other digitaloids in serum. The method is based on the inhibition of Na + , K + --specific ATPase enzyme by digitaloids as a result of receptor binding principle and the measurement of this inhibition with the firefly luceferin--luciferase bioluminescence assay of APT. Method is specific and has similar sensitivity to that of the commonly used radioimmunoassay, but simpler and can be performed with low-cost instrumentation.
Inventors
- 👤 Seppo Kolehmainen
- 👤 Veikko Tarrkkanen
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 08:15
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/34; C12Q1/42; C12Q1/66; C12Q1/008
Claims Analysis
Total Claims: 4
Claim 1
Independent Claim
123 words
A method for measuring the concentration of digoxin and other digitaloids in a sample containing the same by detecting the deactivation thereof on a Na + , K + -specific ATPase enzyme, comprising: incubating said sample, together with a quantity of Na + , K + -specific ATPase enzyme under conditions and for a period sufficient to deactivate a part of said quantity; further incubating the resulting product with a quantity of ATP in excess of that sufficient to be hydrolyzed by said ATPase enzyme and under conditions to promote hydrolysis thereof; and measuring the resulting concentration of ATP by means of the firefly luciferin-luciferase bioluminescent reaction and quantifying such measurement to determine the concentration of digoxin and other disitaloids in the sample.
Claim 2
Independent Claim
123 words
A method for measuring the concentration of digoxin and other digitaloids in a sample containing the same by detecting the deactivation thereof on a Na + , K + -specific ATPase enzyme, comprising: incubating said sample, together with a quantity of Na + , K + -specific ATPase enzyme under conditions and for a period sufficient to deactivate a part of said quantity; further incubating the resulting product with a quantity of ATP in excess of that sufficient to be hydrolyzed by said ATPase enzyme and under conditions to promote hydrolysis thereof; and measuring the resulting concentration of ATP by means of the firefly luciferin-luciferase bioluminescent reaction and quantifying such measurement to determine the concentration of digoxin and other disitaloids in the sample.
Claim 3
Dependent Claim
36 words
A method according to claim 1, wherein the activity of the ATPase enzyme is measured kinetically as a continuous declining light intensity of the bioluminescent reaction while the ATPase enzyme hydrolyzes added ATP during the measurement.
Claim 4
Dependent Claim
49 words
A method according to claim 1, wherein the measured concentration of ATPase enzyme is used to calculate the concentration of digitaloids in the sample by comparing the deactivation of Na + , K + -specific ATPase in the sample to a standard sample containing a known concentration of digitaloids.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1984-04-27 | 1984-10-27 | 1985-04-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1988-04-27 | 1988-10-27 | 1989-04-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1992-04-27 | 1992-10-27 | 1993-04-28 | $7700.00 | Window Closed / Lapsed |