Tissue Factor Stabilization via Surfactant-Nickel Complex
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Solution Overview
Problem
Natural and recombinant tissue factors used in clotting time measurements suffer from poor stability due to impurities and protein denaturation, leading to reduced biological activity over time, and existing stabilization methods can affect clotting time measurements or require costly purification processes.
Innovation Solution
The use of nonionic surfactants such as polyoxyethylene octyl phenyl ether and nickel ions in combination with tissue factors to form a complex, which stabilizes the tissue factor and maintains its biological activity during storage, either at room temperature or in frozen conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If natural tissue factor is extracted from animal brain, then the tissue factor can be obtained as a clotting reagent material, but it contains impurities such as blood components, lipoproteins and plasma proteins that cause poor stability and precipitate formation during long-term storage
Solution Approach 1:
The patent extracts only the necessary tissue factor component from animal brain while removing impurities such as blood components, lipoproteins and plasma proteins through purification processes. This extraction approach obtains sufficient tissue factor quantity while eliminating the sources of instability that cause precipitate formation during storage.
Solution Approach 2:
The patent creates a composite reagent system combining purified tissue factor with stabilizing agents and buffers. This composite formulation maintains tissue factor stability during long-term storage by integrating multiple components that work together to prevent degradation and precipitate formation, resolving the contradiction between obtaining sufficient tissue factor and maintaining storage stability.
2Stability of the object's composition
If stabilizers such as BSA are added to improve tissue factor stability, then storage stability is improved, but the stabilizer may affect clotting time measurements
Solution Approach 1:
The patent introduces inert buffers and stabilizing agents that act as intermediaries between the tissue factor and the measurement system. These intermediaries protect the tissue factor during storage without interfering with the clotting time measurement process, thereby maintaining both storage stability and measurement precision.
Solution Approach 2:
The patent optimizes the concentration and type of stabilizing agents to achieve the minimum effective amount that maintains storage stability without reaching thresholds that would interfere with clotting time measurements. By carefully controlling these parameters, both stability and measurement accuracy are preserved.
3Stability of the object's composition
If partial purification is performed by antibody column chromatography or gel filtration chromatography, then stability is improved, but time and costs are required for preparing a large amount of tissue factor
Solution Approach 1:
The patent performs preliminary purification steps during the tissue factor extraction process itself, removing major impurities before storage and use. This preliminary action reduces the need for time-consuming additional purification steps later, achieving both improved stability and reduced preparation time.
Solution Approach 2:
The patent extracts and removes impurities such as blood components, lipoproteins and plasma proteins during the initial preparation phase through efficient purification methods. This extraction approach achieves sufficient purification for stability improvement without requiring extensive additional purification steps that would consume excessive time and resources.
4Quantity of substance
If genetic recombinant tissue factor is used, then purity is improved, but the solution may be liable to denature the protein and reduce biological activity depending on storage conditions
Solution Approach 1:
The patent formulates genetic recombinant tissue factor into a composite system containing stabilizing agents, buffers, and protective components. This composite formulation protects the pure recombinant protein from denaturation during storage, maintaining both the high purity achieved through recombinant production and the biological activity reliability needed for consistent measurement results.
Solution Approach 2:
The patent optimizes storage conditions and formulation parameters such as pH, ionic strength, and stabilizer concentration to prevent protein denaturation of genetic recombinant tissue factor. By carefully controlling these parameters, the patent maintains both the high purity of recombinant tissue factor and its biological activity stability across various storage conditions.
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
The combination of nonionic surfactants and nickel ions effectively maintains the biological activity of tissue factors for a longer period, improving storage stability without affecting clotting time measurements, even in frozen storage, and enhancing the reagent's stability and usability for clinical applications.
Implementation Method 1
The use of nonionic surfactants such as polyoxyethylene octyl phenyl ether and nickel ions in combination with tissue factors to form a complex, which stabilizes the tissue factor
Implementation Method 2
nonionic surfactants such as polyoxyethylene octyl phenyl ether and nickel ions in combination with tissue factors to form a complex
Implementation Method 3
The addition of a chelating agent such as EDTA to a solution containing a protein such as an enzyme is known to remove the influence of a heavy metal
Implementation Method 4
The peroxidation, oxidation and denaturation of a protein by the catalytic action of a heavy metal ion can be prevented by the chelating agent
Data Source
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AI summary
The present invention provides a reagent for measuring clotting time comprising a nonionic surfactant, a nickel ion and a tissue factor. A method for stabilizing a tissue factor, and use of a nonionic surfactant and a nickel salt for stabilizing a tissue factor are also disclosed.