US4334018A
Expired / Lapsed
US
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Process and reagent for the determination of prothrombin
Assignee: Roche Diagnostics GmbH
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Filing: 1980-12-15
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Granted: 1982-06-08
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Expires: 2000-12-15
Abstract
Abstract A novel process for the determination of prothrombin in biological material, by conversion of prothrombin into thrombin, enzymatic fission of a thrombin substrate and measurement of a fission product, the improvement comprising incubating the test solution with the addition of Factor Xa.
Inventors
- 👤 Bruno Kirchhof
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 11:17
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/56; C07; C07K; C07K5/00; C07K5/04; C07K5/08; C07K5/0802; C07K5/0804; C07K5/0806; C07K5/0812; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12Q2337/00; C12Q2337/10; C12Q2337/12; G; G01; G01N; G01N2333/00; G01N2333/435; G01N2333/46; G01N2333/4609; G01N2333/4613; G01N2333/4616; G01N2333/4633; G01N2333/90; G01N2333/914; G01N2333/948; G01N2333/95; G01N2333/964; G01N2333/96425; G01N2333/96427; G01N2333/9643; G01N2333/96433; G01N2333/96441; G01N2333/96444; G01N2333/974; G01N2405/00; G01N2405/04
Claims Analysis
Total Claims: 29
Claim 1
Independent Claim
48 words
In a process for the determination of prothrombin in biological material, by conversion of prothrombin into thrombin, enzymatic fission of a thrombin substrate and measurement of product, the improvement comprising incubating the test solution with the addition of Factor Xa as the sole enzyme activating prothrombin into thrombin.
Claim 2
Dependent Claim
48 words
In a process for the determination of prothrombin in biological material, by conversion of prothrombin into thrombin, enzymatic fission of a thrombin substrate and measurement of product, the improvement comprising incubating the test solution with the addition of Factor Xa as the sole enzyme activating prothrombin into thrombin.
Claim 3
Dependent Claim
12 words
Process as claimed in claim 1, wherein human Factor Xa is added.
Claim 4
Dependent Claim
51 words
Process as claimed in claim 1, wherein the Factor Xa is produced by treating plasma with a prothrombin activator, centrifuging, mixing the supernatant with a protein adsorbent, eluting the precipitate with a protein elution agent and mixing the eluate with a Factor X activator and optionally with a soluble calcium salt.
Claim 5
Dependent Claim
11 words
Process as claimed in claim 3, wherein human plasma is used.
Claim 6
Dependent Claim
15 words
Process as claimed in claim 1 wherein the thrombin substrate is a synthetic peptide-p-nitroanilide derivative.
Claim 7
Dependent Claim
16 words
Process as claimed in claim 5, wherein the thrombin substrate is N-Tos-Gly-Pro-Arg-pNA, N-Cbz-Gly-Pro-Arg-pNA, H-D-Phe-Pip-Arg-pNA or Bz-Phe-Val-Arg-pNA.
Claim 8
Dependent Claim
13 words
Process as claimed in claim 1, wherein the biological material is blood plasma.
Claim 9
Dependent Claim
36 words
Reagent for the determination of prothrombin in biological material, comprising a thrombin substrate and a prothrombin activator, wherein the prothrombin activator is Factor Xa in a concentration of from 0.2 to 2.5% of the biological material.
Claim 10
Dependent Claim
36 words
Reagent for the determination of prothrombin in biological material, comprising a thrombin substrate and a prothrombin activator, wherein the prothrombin activator is Factor Xa in a concentration of from 0.2 to 2.5% of the biological material.
Claim 11
Dependent Claim
14 words
Reactant as claimed in claim 8 wherein the prothrombin activator is human Factor Xa.
Claim 12
Dependent Claim
51 words
Reagent as claimed in claim 8 wherein the Factor Xa is prepared by treating plasma with a prothrombin activator, centrifuging, mixing the supernatant with a protein adsorbent, eluting the precipitate with a protein elution agent and mixing the eluate with a Factor X activator and optionally with a soluble calcium salt.
Claim 13
Dependent Claim
15 words
Reagent as claimed in claim 10 wherein human plasma is used as the biological material.
Claim 14
Dependent Claim
49 words
Reagent for the determination of prothrombin in biological material, comprising a buffer selected from the group consisting of tris and imidazole buffers, Factor Xa, co-reagents for Factor Xa and a synthetic thrombin substrate said Factor Xa being in a concentration of from 0.2 to 2.5% of the biological material.
Claim 15
Dependent Claim
49 words
Reagent for the determination of prothrombin in biological material, comprising a buffer selected from the group consisting of tris and imidazole buffers, Factor Xa, co-reagents for Factor Xa and a synthetic thrombin substrate said Factor Xa being in a concentration of from 0.2 to 2.5% of the biological material.
Claim 16
Dependent Claim
12 words
Reagent as claimed in claim 12 wherein human Factor Xa is used.
Claim 17
Dependent Claim
51 words
Reagent as claimed in claim 12 wherein the Factor Xa is prepared by treating plasma with a prothrombin activator, centrifuging, mixing the supernatant with a protein adsorbent, eluting the precipitate with a protein elution agent and mixing the eluate with a Factor X activator and optionally with a soluble calcium salt.
Claim 18
Dependent Claim
11 words
Reagent as claimed in claim 14 wherein human plasma is used.
Claim 19
Dependent Claim
14 words
Reagent as claimed in claim 12 wherein the coreagents are phospholipids and calcium chloride.
Claim 20
Dependent Claim
17 words
Reagent as claimed in claim 12 wherein the synthetic thrombin substrate is N-Tos-Gly-Pro-Arg-pNA, H-D-Phe-Pip-Arg-pNA, N-Cbz-Gly-Pro-Arg-pNA or Bz-Phe-Val-Arg-pNA.
Claim 21
Dependent Claim
39 words
Reagent as claimed in claim 16 comprising 0.3 to 6.5 g./ml. of phospholipids, 0.7 to 10 mM/liter calcium chloride and 150 to 380 M/liter thrombin substrate, the Factor Xa concentration corresponding to 0.2 to 2.5% of the plasma extract.
Claim 22
Dependent Claim
50 words
In a process for the determination of prothrombin in biological material, by conversion of prothrombin into thrombin, enzymatic fission of a thrombin substrate and measurement of product, the improvement comprising incubating the test solution with Factor Xa in a concentration of from 0.2 to 2.5% of the biological material extract.
Claim 23
Dependent Claim
50 words
In a process for the determination of prothrombin in biological material, by conversion of prothrombin into thrombin, enzymatic fission of a thrombin substrate and measurement of product, the improvement comprising incubating the test solution with Factor Xa in a concentration of from 0.2 to 2.5% of the biological material extract.
Claim 24
Dependent Claim
12 words
Improvement as claimed in claim 19 wherein human Factor Xa is added.
Claim 25
Dependent Claim
49 words
Improvement as claimed in claim 19 wherein Factor Xa is produced by treating plasma with a prothrombin activator, centrifuging, mixing the supernatant with a protein adsorbent, eluting the precipitate with a protein elution agent and mixing the eluate with Factor X activator and optionally with a soluble calcium salt.
Claim 26
Dependent Claim
11 words
Improvement as claimed in claim 21 wherein human plasma is used.
Claim 27
Dependent Claim
15 words
Improvement as claimed in claim 19 wherein the thrombin substrate is a synthetic peptide-nitroanilide derivative.
Claim 28
Dependent Claim
16 words
Improvement as claimed in claim 19 wherein the thrombin substrate is N-Tos-Gly-Pro-Arg-pNA, N-Cbz-Gly-Pro-Arg-pNA, H-D-Phe-Pip-Arg-pNA, or Bz-Phe-Val-Arg-pNA.
Claim 29
Dependent Claim
13 words
Improvement as claimed in claim 19 wherein the biological material is blood plasma.
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) | 1985-06-07 | 1985-12-07 | 1986-06-08 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1989-06-07 | 1989-12-07 | 1990-06-08 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1993-06-07 | 1993-12-07 | 1994-06-08 | $7700.00 | Window Closed / Lapsed |