US4234682A
Expired / Lapsed
US
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Method and reagent for determining biologically active heparin in plasma
Assignee: Roche Diagnostics GmbH
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Filing: 1979-02-21
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Granted: 1980-11-18
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Expires: 1999-02-21
Abstract
Abstract A method for determining biologically active heparin comprising adding, in the absence of exogenous antithrombin III, a proteolytic enzyme selected from the group consisting of thrombin and factor Xa and a chromogen substrate for the enzyme to a heparin sample, measuring the dye released from the chromogen substrate and thus determining the amount of heparin present.
Inventors
- 👤 Knut Bartl
- 👤 Helmut Lill
- 👤 Joachim Ziegenhorn
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 23:52
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/56; C12Q2337/00; C12Q2337/10; C12Q2337/12; G; G01; G01N; G01N2333/00; G01N2333/81; G01N2333/8107; G01N2333/811; G01N2333/8114; G01N2333/8117; G01N2333/90; G01N2333/914; G01N2333/948; G01N2333/95; G01N2333/964; G01N2333/96425; G01N2333/96427; G01N2333/9643; G01N2333/96433; G01N2333/96441; G01N2333/96444; G01N2333/974; G01N2400/00; G01N2400/10; G01N2400/38; G01N2400/40; Y; Y10; Y10T; Y10T436/00; Y10T436/10; Y10T436/108331; Y10T436/14; Y10T436/142222; Y10T436/143333
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
61 words
Method for the determination of the biological activity of heparin in plasma which method comprises adding to the heparin sample a proteolytic enzyme selected from thrombin or factor Xa, and a chromogen substrate of said proteolytic enzyme, without addition of exogenous antithrombin III, and measuring the dye released from the chromogen substrate as a measure of the heparin activity initially present.
Claim 2
Dependent Claim
61 words
Method for the determination of the biological activity of heparin in plasma which method comprises adding to the heparin sample a proteolytic enzyme selected from thrombin or factor Xa, and a chromogen substrate of said proteolytic enzyme, without addition of exogenous antithrombin III, and measuring the dye released from the chromogen substrate as a measure of the heparin activity initially present.
Claim 3
Dependent Claim
29 words
Method as claimed in claim 1 wherein said chromogen substrate is a peptide having a p-nitroanilide moiety bound, via amide bonding, to the carboxyl group of an arginine moiety.
Claim 4
Dependent Claim
23 words
Method as claimed in claim 1 wherein said proteolytic enzyme is thrombin and said chromogen substrate is selected from Bz-Phe-Val-Arg-pNA, H-D-Phe-Pip-Arg-pNA and Tos-Gly-Pro-Arg-pNA.
Claim 5
Dependent Claim
19 words
Method as claimed in claim 1 wherein the proteolytic enzyme is factor Xa and said chromogen substrate is Bz-ILe-Glu-Gly-Arg-pNA.
Claim 6
Dependent Claim
14 words
Method as claimed in claim 1 wherein aprotinin is additionally added in said determination.
Claim 7
Dependent Claim
61 words
Reagent for the determination of heparin in plasma consisting essentially of 0.01 to 0.3 moles/l of buffer, pH 6 to 9 0.01 to 0.25 moles/l of alkali chloride, 200 to 1100 NIH/l of thrombin 0.05 to 10 mmoles/l of chromogen substrate for thrombin, 0 to 0.01 g/l of aprotinin, 0 to 10 g/l of polyethylene glycol. 0 to 0.03 moles/l EDTA
Claim 8
Dependent Claim
61 words
Reagent for the determination of heparin in plasma consisting essentially of 0.01 to 0.3 moles/l of buffer, pH 6 to 9 0.01 to 0.25 moles/l of alkali chloride, 200 to 1100 NIH/l of thrombin 0.05 to 10 mmoles/l of chromogen substrate for thrombin, 0 to 0.01 g/l of aprotinin, 0 to 10 g/l of polyethylene glycol. 0 to 0.03 moles/l EDTA
Claim 9
Dependent Claim
63 words
Reagent for the determination of heparin in plasma consisting essentially of 0.01 to 0.3 moles/l of buffer, pH 6 to 9 0.01 to 0.25 moles/l of alkali chloride, 200 to 1100 NIH/l of factor Xa, 0.05 to 10 mmoles/l of chromogen substrate for factor Xa, 0 to 0.01 g/l of aprotinin, 0 to 10 g/l of polyethylene glycol. 0 to 0.03 moles/l EDTA
Claim 10
Dependent Claim
63 words
Reagent for the determination of heparin in plasma consisting essentially of 0.01 to 0.3 moles/l of buffer, pH 6 to 9 0.01 to 0.25 moles/l of alkali chloride, 200 to 1100 NIH/l of factor Xa, 0.05 to 10 mmoles/l of chromogen substrate for factor Xa, 0 to 0.01 g/l of aprotinin, 0 to 10 g/l of polyethylene glycol. 0 to 0.03 moles/l EDTA
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1983-11-18 | 1984-05-19 | 1984-11-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-11-18 | 1988-05-19 | 1988-11-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-11-18 | 1992-05-19 | 1992-11-18 | $7700.00 | Window Closed / Lapsed |