A method for the simultaneous measurement of proteolylic enzyme generation and clot strength in plasma or whole blood or any appropriate biological sample derived from blood. The measurement method encompasses the use of a detectable substrate which includes a moiety that can be released upon reaction with the targeted proteolytic enzyme, and elements for measurement of an increase in viscosity of clot strength.
C12Q 1/56 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
G01N 11/14 - Investigating flow properties of materials, e.g. viscosity or plasticityAnalysing materials by determining flow properties by moving a body within the material by using rotary bodies, e.g. vane
G01N 33/49 - Physical analysis of biological material of liquid biological material blood
G01N 33/86 - Chemical analysis of biological material, e.g. blood, urineTesting involving biospecific ligand binding methodsImmunological testing involving blood coagulating time
G01N 33/58 - Chemical analysis of biological material, e.g. blood, urineTesting involving biospecific ligand binding methodsImmunological testing involving labelled substances
2.
COMPOUNDS AND METHODS FOR INHIBITION OF BINDING OF ICAM-4 TO PLATELET INTEGRIN αIIbβ3
The present invention relates to compounds and methods for inhibition of binding of ICAM-4 to platelet integrin αIIbβ3. The present invention also relates to a screening method and to a kit to detect or monitor in vitro the effect of a substance, drug or pharmaceutical agent on the ICAM-4/ αIIbβ3 interaction in a biological sample, while simulating blood flow conditions existing in vivo. It concerns in particular an anti-ICAM-4 antibody or a fragment thereof capable of blocking ICAM-4 binding to platelet integrin αIIbβ3, or an ICAM-4 mimetic peptide blocking ICAM-4 binding to platelet integrin αIIbβ3, or an anti-CD61 antibody or a fragment thereof capable of blocking ICAM-4 binding to platelet integrin αIIbβ3 for use in the treatment of a thrombotic disease, and for the preparation of pharmaceutical compositions for such treatment.
C07K 16/28 - Immunoglobulins, e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
C07K 7/08 - Linear peptides containing only normal peptide links having 12 to 20 amino acids
C07K 11/00 - Depsipeptides having up to 20 amino acids in a fully defined sequenceDerivatives thereof
The present invention relates to the simultaneous measurement of proteolylic enzyme generation and clot strength in plasma or whole blood or any appropriate biological sample derived from blood. The measurement method encompasses the use of a detectable substrate which comprises a moiety that can be released upon reaction with the targeted proteolytic enzyme, and means for measurement of an increase in viscosity of clot strength.
C12Q 1/56 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
G01N 33/86 - Chemical analysis of biological material, e.g. blood, urineTesting involving biospecific ligand binding methodsImmunological testing involving blood coagulating time
G01N 11/14 - Investigating flow properties of materials, e.g. viscosity or plasticityAnalysing materials by determining flow properties by moving a body within the material by using rotary bodies, e.g. vane
G01N 33/49 - Physical analysis of biological material of liquid biological material blood
4.
THERMOSTABLE INHIBITORS OF ACTIVATION OF THE BLOOD CLOTTING SYSTEM THROUGH CONTACT WITH FOREIGN SURFACES.
The present invention relates to the field of blood clotting. Specifically, the invention relates to particular inhibitors of artificial activation of the blood clotting process through contact with foreign surfaces.
Disclosed is a method for measuring thrombin generation in a whole blood sample. The whole blood sample may be applied forthwith, without prior processing. The blood cells and blood plasma in the whole blood sample are separated by (lateral) flow migration. Also disclosed is an assembly of a sample support and a device dedicated to measure thrombin generation in a whole blood sample. Advantageously, the sample support comprises a separator medium allowing separation of whole blood into blood cells and blood plasma by means of (lateral) flow migration.
The invention relates to a method for in vitro determining thrombin activity in a sample wherein the sample is a blood sample and thrombin generation is measured by the steps of: —contacting a layer of said sample with a fluorogenic substrate of thrombin, wherein said layer has a thickness within a range of 0.05 to 5 mm and a surface within a range of 10 to 500 mm2; —allowing thrombin to generate in said sample; —measuring the fluorescence emitted from the surface of the layer, by the fluorescent group released from the fluorogenic substrate as a result of enzymatic action of generated thrombin on said fluorogenic substrate.
The invention relates to time-course measurement of enzymatic activity corrected for impacts of disturbances relating to the reaction of the enzyme with a substrate. The in vitro method of the invention is especially intended for the measurement of clotting or the fibrinolysis activity in vitro. The invention enables to measure the exact time course of an enzymatic activity that develops and/or disappears in a reaction mixture, especially in a blood sample. The invention thus relates to a method of determination of the course of an enzyme activity that is variable in time, wherein said activity is probed by conversion of a substrate of the enzyme, comprising, in a selected test set up and for a determined substrate of the enzyme, the determination of the velocity of signal production (dFdiag/dt) resulting from a time curve of the signal (Fdiag=f(A)) obtained from splitting said substrate when it is contacted with a determined initially fixed concentration of the enzyme (E) and providing a 'diagnostic plot' with the values of (dFdiag/dt) against the signal (Fdiag) and determining whether said diagnostic plot is either a straight line or a parabola and in the same test conditions, for a given test sample, the determination of the signal production (Fexp) resulting from splitting the substrate by the enzyme generating in and/or disappearing from the sample and providing the time curve of signal Fexp=f(t); and transforming the obtained experimental value of the signal (Fexp) into an ideal value (Ftransf).
G01N 33/86 - Chemical analysis of biological material, e.g. blood, urineTesting involving biospecific ligand binding methodsImmunological testing involving blood coagulating time
C12Q 1/37 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving hydrolase involving peptidase or proteinase
C12Q 1/56 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
8.
Diagnostic test for determining the concentration of transient proteolytic activity in composite biological media
A method is provided for determining in real time the course of thrombin activity in a sample of blood or plasma as it appears in and disappears from the simple which comprises adding a thrombin substrate to the sample that, per unit time, produces a detectable signal in a quantity that bears relation to the amount of thrombin present. Simultaneously, in a control sample of the same blood or plasma in which thrombin generation is not triggered, the activity of a standard preparation with invariable thrombin activity is measured. The exact molar amount of thrombin present at any moment is obtained by comparison of the activity measured in clotting blood and the simultaneously measured calibrator. The method is useful inter alia for diagnosing hyper- and hypo-coaguable states, either congenital, acquired or drug-induced in humans and animals. Also provided is a kit for use in this method.
