Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: A method of determining the course of enzyme activity that is variable in time, wherein the activity is probed by conversion of a substrate of the enzyme, includes, in a selected test set up and for a determined substrate of the enzyme, determining the velocity of signal production (dFdiag/dt) resulting from a time curve of the signal (Fdiao=f(A)) obtained from splitting the 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 the diagnostic plot is either a straight line or a parabola and in the same test conditions, for a given test sample, determining 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).

Abstract: A method of determining the course of enzyme activity that is variable in time, wherein the activity is probed by conversion of a substrate of the enzyme, includes, in a selected test set up and for a determined substrate of the enzyme, determining the velocity of signal production (dFdiag/dt) resulting from a time curve of the signal (Fdiao=f(A)) obtained from splitting the 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 the diagnostic plot is either a straight line or a parabola and in the same test conditions, for a given test sample, determining 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).

Abstract: 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.

Abstract: 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.

Abstract: A process is provided for the assessment of an active proteolytic enzyme in a blood or another biological fluid sample possibly comprising a complex of said proteolytic enzyme and &agr;2-macroglobulin, wherein said sample is contacted with a substrate comprising a molecule of sufficient size coupled to a signal-substrate, said signal-substrate comprising a detectable leaving group, wherein said substrate is hydrolysed by said proteolytic enzyme but not by said complex. The proteolytic enzyme is preferably selected from the group consisting of thrombin, activated clotting factor, activated fibrinolytic factor, and activated component of the complement system. Of these, thrombin is most preferred. The molecules of sufficient size are preferably water soluble and selected from the group consisting of inert protein, preferably ovalbumin, polysaccharide, and synthetic polymer. The size of these molecules is such that they will not fit into the cavity of the &agr;2M molecules.