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US4563420A
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Process for assaying the activity of tissue plasminogen activator, and kit suitable for use in said process
Assignee: NEDERLANDSE CENTRALE ORGANISATIE VOOR TOEGEPAST-NATUURWETENSCHAPPELIJK ONDERZOEK JULIANA VAN STOLBERGLAAN 148 2595 CL HAGUE
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Filing: 1983-05-06
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Granted: 1986-01-07
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Expires: 2003-05-06
Abstract
Abstract The invention relates to a process for assaying the activity of tissue plasminogen activator (TA), in which a sample is incubated with plasminogen, a substrate for plasmin giving a detectable reaction product on reaction with plasmin, preferably H-D-Val-Leu-Lys-pNA, and, as a stimulator for the conversion of plasminogen to plasmin by TA, water-soluble fibrinogen or fibrin fragments comprising at least partially the D-domains of the fibrinogen or fibrin molecule. The amount of the detectable reaction product of the plasmin substrate is measured. Preferably, said fibrinogen or fibrin fragments have been obtained by reacting fibrinogen or fibrin with CNBr and removing lower molecular components from the reaction product. The invention also relates to kits for performing the assay and comprising the reagents to be used.
Inventors
- 👤 Johan H. Verheijen
- 👤 Willem Nieuwenhuizen
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 13:02
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/56; C12Q2337/00; C12Q2337/10; C12Q2337/12; G; G01; G01N; G01N2333/00; G01N2333/435; G01N2333/745; G01N2333/75; G01N2333/90; G01N2333/914; G01N2333/948; G01N2333/968; G01N2333/972; G01N2333/9726
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
96 words
In a process for assaying the activity of tissue plasminogen activator in a sample, comprising the incubation of the sample with: (a) plasminogen, (b) a stimulator for the conversion of plasminogen to plasmin by tissue plasminogen activator, and (c) a substrate for plasmin which gives a detectable reaction product on reaction with plasmin; followed by a step of measuring the amount of reaction product formed, the improvement comprising using, as the stimulator (b), water-soluble fibrinogen or fibrin fragments comprising at least partially the D-domains of the fibrinogen or fibrin molecule and being smaller than fragment X.
Claim 2
Dependent Claim
96 words
In a process for assaying the activity of tissue plasminogen activator in a sample, comprising the incubation of the sample with: (a) plasminogen, (b) a stimulator for the conversion of plasminogen to plasmin by tissue plasminogen activator, and (c) a substrate for plasmin which gives a detectable reaction product on reaction with plasmin; followed by a step of measuring the amount of reaction product formed, the improvement comprising using, as the stimulator (b), water-soluble fibrinogen or fibrin fragments comprising at least partially the D-domains of the fibrinogen or fibrin molecule and being smaller than fragment X.
Claim 3
Dependent Claim
32 words
The process of claim 1, wherein the fibrinogen or fibrin fragments have been obtained by reacting fibrinogen or fibrin with CNBr and removing the lower molecular weight components from the reaction product.
Claim 4
Dependent Claim
77 words
A kit for assaying the activity of tissue plasminogen activator in a sample, comprising: (a) a container with a measured amount of a substrate for plasmin giving a detectable reaction product on reaction with plasmin, (b) a container with a measured amount of plasminogen, and (c) a container with a measured amount of water soluble fibrinogen or fibrin fragments comprising at least partially the D-domains of the fibrinogen or fibrin molecule and being smaller than fragment X.
Claim 5
Dependent Claim
77 words
A kit for assaying the activity of tissue plasminogen activator in a sample, comprising: (a) a container with a measured amount of a substrate for plasmin giving a detectable reaction product on reaction with plasmin, (b) a container with a measured amount of plasminogen, and (c) a container with a measured amount of water soluble fibrinogen or fibrin fragments comprising at least partially the D-domains of the fibrinogen or fibrin molecule and being smaller than fragment X.
Claim 6
Dependent Claim
13 words
A kit according to claim 3, wherein the substrate for plasmin is H-D-Val-Leu-Lys-p-NA.
Claim 7
Dependent Claim
40 words
A kit according to claim 3, wherein the fibrinogen or fibrin fragments have been obtained by reaction of fibrinogen or fibrin with CNBr, removal of the lower molecular weight components from the reaction product, and lyophilization of the solution obtained.
Claim 8
Dependent Claim
47 words
A kit according to claim 3, wherein the substrate for plasmin is H-D-Val-Leu-Lys-p-NA, and the fibrinogen or fibrin fragments have been obtained by reaction of fibrinogen or fibrin with CNBr, removal of the lower molecular weight components from the reaction product and lyophilization of the solution obtained.
Claim 9
Dependent Claim
19 words
A kit according to claim 6, wherein the weight ratio of H-D-Val-Leu-Lys-p-NA to plasminogen to fibrinogen fragments is 12-20:1:10-20.
Claim 10
Dependent Claim
19 words
A kit according to claim 6, wherein the weight ratio of H-D-Val-Leu-Lys-p-NA to plasminogen to fibrinogen fragments is 14-15:1:10.
Claim 11
Dependent Claim
18 words
A kit according to claim 3, additionally comprising a container with a buffer solution optionally containing a surfactant.
Claim 12
Dependent Claim
16 words
A kit according to claim 3, wherein the buffer solution is 0.1M Tris-HCl buffer pH 7.5.
Claim 13
Dependent Claim
12 words
A kit according to claim 9, wherein the surfactant is Tween 80.
Claim 14
Dependent Claim
22 words
A kit according to claim 9, wherein the buffer solution is 0.1M Tris-HCl buffer pH 7.5, and the surfactant is Tween 80.
Claim 15
Dependent Claim
18 words
A kit according to claim 3, additionally comprising a container with a measured amount of tissue plasminogen activator.
Claim 16
Dependent Claim
17 words
A kit according to claim 3, additionally comprising measured amounts of anti-TA-IgG and anti-UK-IgG in separate containers.
References Cited (Backward Citations)
US4011142A
Cited
US4278762A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1989-01-06 | 1989-07-08 | 1990-01-07 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-01-06 | 1993-07-08 | 1994-01-07 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-01-06 | 1997-07-08 | 1998-01-07 | $7700.00 | Window Closed / Lapsed |