EP0094720A1
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Process for assaying the activity of tissue plasminogen activator, and kit suitable for use in said process
Assignee: Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO
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Filing: 1983-05-09
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Granted: —
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Expires: 2003-05-09
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 Hendrikus Verheijen
- 👤 Willem Nieuwenhuizen
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-01 08:05
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: 13
Claim 1
Independent Claim
101 words
A process for assaying the activity of tissue plasminogen activator (TA) in a sample, in which a sample is incubated with plasminogen, a stimulator for the conversion of plasminogen to plasmin by tissue plasminogen activator, as well as a substrate for plasmin giving a detectable reaction product on reaction with plasmin, and the amount of said reaction product formed is measured, characterized by using 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 as a stimulator of the conversion of plasminogen to plasmin by tissue plasminogen activator.
Claim 2
Dependent Claim
101 words
A process for assaying the activity of tissue plasminogen activator (TA) in a sample, in which a sample is incubated with plasminogen, a stimulator for the conversion of plasminogen to plasmin by tissue plasminogen activator, as well as a substrate for plasmin giving a detectable reaction product on reaction with plasmin, and the amount of said reaction product formed is measured, characterized by using 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 as a stimulator of the conversion of plasminogen to plasmin by tissue plasminogen activator.
Claim 3
Dependent Claim
30 words
The process of claim 1, characterized by using fibrinogen or fibrin fragments obtained by reacting fibrinogen or fibrin with CNBr and removing the lower molecular components from the reaction product.
Claim 4
Dependent Claim
81 words
Kit for carrying out the process according to claim 1 or 2 and comprising at least the following components: 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
13 words
Kit according to claim 3 in which the substrate for plasmin is H-D-Val-Leu-Lys-p-NA.
Claim 6
Dependent Claim
41 words
Kit according to claim 3 or 4, characterized in that the fibrin or fibrinogen fragments have been obtained by reaction of fibrin or fibrinogen with CNBr, removal of the lower molecular components from the reaction product, and lyophilizing the solution obtained.
Claim 7
Dependent Claim
47 words
Kit according to any one of claims 3-5 comprising a) a container with H-D-Val-Leu-Lys-p-NA b) a container with plasminogen, and c) a container with fibrinogen fragments according to claim 5, the weight ratio of said H-D-Val-Leu-Lys-p-NA to said plasminogen to said fibrinogen fragments being 12-20:1:10-20, particularly 14-15:1:10.
Claim 8
Dependent Claim
20 words
Kit according to any one of claims 3-6, additionally comprising a container with a buffer solution optionally containing a surfactant.
Claim 9
Dependent Claim
17 words
Kit according to claim 7, in which the buffer solution is 0.1 M Tris-HC1 buffer pH 7.5.
Claim 10
Dependent Claim
14 words
Kit according to claim 7 or 8, in which the surfactant is Tween 80.
Claim 11
Dependent Claim
15 words
Kit according to any one of claims 3-9 additionally comprising one or more operating instructions.
Claim 12
Dependent Claim
21 words
Kit according to any one of claims 3-10 additionally comprising a container with a measured amount of tissue plasminogen activator (TA).
Claim 13
Dependent Claim
18 words
Kit according to any one of claims 3-11 additionally comprising containers with measured amounts of anti-TA-IgG and anti-UK-IgG.
References Cited (Backward Citations)
US4214049A
Cited
GB1589738A
Cited
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