Recombinant Tissue Factor Reagent for Clotting Time Analysis
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Solution Overview
Problem
The production of recombinant tissue factor for coagulation tests is hindered by high costs, labor-intensive purification processes, and potential interference with coagulation activity due to the use of bovine-derived materials and purification methods like affinity chromatography, which also risks contamination from host-derived impurities.
Innovation Solution
A reagent for measuring clotting time is developed using a recombinant tissue factor produced in insect or cultured insect cells, combined with a phospholipid, which simplifies the purification process and reduces costs by leveraging genetic engineering techniques and baculovirus expression systems, eliminating the need for bovine-derived materials and minimizing the impact on coagulation activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If recombinant tissue factor is produced using conventional purification methods (affinity chromatography), then purity is improved, but production cost and time increase significantly
Solution Approach 1:
The patent extracts only the essential functional component (tissue factor protein) from the complex bovine cerebrum matrix through selective solubilization and precipitation steps, separating it from non-essential components that complicate purification. This extraction approach obtains sufficient purity for coagulation testing without requiring elaborate chromatography systems.
Solution Approach 2:
The invention employs simple, inexpensive purification steps (solvent extraction, precipitation, filtration) that can be performed with basic laboratory equipment rather than expensive, complex chromatography systems. These simple methods are sufficient for the application's needs and can be easily implemented in routine production.
2Reliability
If bovine cerebrum is used as starting material, then coagulation activity is maintained, but risk of BSE contamination increases
Solution Approach 1:
The patent extracts tissue factor from bovine cerebrum under controlled conditions and immediately processes it through purification steps that separate the functional protein from potential contaminants. The extracted tissue factor is then formulated into a reagent that maintains coagulation activity while minimizing exposure to BSE risks through controlled handling and processing.
Solution Approach 2:
The invention uses an intermediary processing system where tissue factor is extracted, purified, and formulated into a stable reagent product. This intermediary formulation process creates a controlled barrier between the starting material and the final product, ensuring that only the essential coagulation-active components remain while potential contaminants are eliminated or inactivated.
3Manufacturing precision
If affinity chromatography with immobilized monoclonal antibody is used for purification, then purity is improved, but manufacturing cost increases
Solution Approach 1:
The patent extracts tissue factor using simple solvent-based methods and precipitation techniques that rely on the protein's inherent solubility properties rather than requiring expensive antibody-based affinity chromatography. This extraction approach achieves sufficient purity for coagulation testing at a fraction of the cost of monoclonal antibody methods.
Solution Approach 2:
The invention employs inexpensive, single-use purification methods (ammonium sulfate precipitation, organic solvent extraction, filtration) that can be performed with basic equipment. These disposable-style simple methods eliminate the need for expensive, complex affinity chromatography systems while providing adequate purity for the intended application.
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 solution provides a reagent with coagulation activity and sensitivity comparable to conventional thromboplastin-based reagents, while reducing production costs and simplifying the purification process, thus enhancing productivity and safety by avoiding BSE risks.
Implementation Method 1
Thromboplastin is a complex between a protein called tissue factor and phospholipid. Thromboplastin is involved in blood coagulation.
Data Source
AI summary
A reagent for measuring clotting time is described. The reagent comprises a complex of a phospholipid and a recombinant tissue factor obtained by using an insect or a cultured insect cell as a host; and a soluble component derived from the insect or the cultured insect cell.A method for manufacturing the reagent is also described.