Real-Time Thrombin Activity Measurement via Parallel Reference Sample
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Solution Overview
Problem
Current methods for determining thrombin activity in blood or plasma are labor-intensive, insensitive to mild hemostatic disorders, and cannot accurately measure thrombin activity in real time due to limitations in substrate consumption, optical density measurements, and interference from fibrinogen and platelets.
Innovation Solution
A method involving the addition of a protease activator and a signal substrate to a sample, with a parallel sample containing a constant known stable proteolytic activity, allows for real-time monitoring of thrombin concentration by comparing the signal development curves to derive the course of thrombin activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If optical density measurement is used to monitor thrombin activity, then continuous real-time monitoring is possible, but the measurement becomes insensitive to mild hemostatic disorders and is interfered with by fibrinogen and platelets
Solution Approach 1:
The patent extracts the measurement from the interfering environment by using a parallel sample system. The test sample undergoes physiological processing with fibrinogen and platelets present, while the reference sample is treated identically except thrombin generation is blocked. This removes the interference factors from the measurement comparison while preserving the thrombin-specific signal.
Solution Approach 2:
The patent introduces a reference sample as an intermediary that mediates the measurement process. This reference sample serves as a control that experiences the same interfering conditions (fibrinogen, platelets, optical properties) but without thrombin generation, allowing the true thrombin signal to be isolated by comparison.
2Duration of action of moving object
If substrate is added to monitor thrombin activity in real time, then continuous monitoring is achieved, but substrate consumption limits the accuracy and duration of measurement
Solution Approach 1:
The patent implements feedback by continuously comparing the test sample signal with the reference sample signal. This feedback mechanism compensates for substrate consumption over time, as both samples consume substrate at similar rates, allowing the differential signal to remain accurate throughout the monitoring duration.
Solution Approach 2:
The patent ensures continuous useful action by maintaining both test and reference samples throughout the entire monitoring period. Rather than taking discrete measurements, the system continuously monitors both samples simultaneously, ensuring that substrate consumption effects are consistently captured and differentiated.
3Adaptability or versatility
If clotting tests are performed to assess hemostatic function, then overall coagulation status is evaluated, but the tests are insensitive to mild hemostatic disorders and require adaptation for specific uses
Solution Approach 1:
The patent segments the assessment into two distinct components: the test sample that captures the full physiological response including thrombin generation, and the reference sample that captures only the background signal. This segmentation allows the differential measurement to specifically isolate thrombin activity, providing sensitivity to mild disorders while maintaining overall system versatility.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach provides a continuous, accurate representation of thrombin activity in real time, enabling the measurement of crucial parameters like lag time and peak height, which are essential for assessing the hemostatic-thrombotic system's function and monitoring antithrombotic drug effects.
Implementation Method 1
adding a protease activator and a signal substrate to a sample, with a parallel sample containing a constant known stable proteolytic activity
Implementation Method 2
adding a signal substrate to step a), said signal substrate causing a detectable signal related to the amount of conversion product formed upon reaction by the generated proteolytic activity
Data Source
AI summary
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.