Factor XIII Purification Using Diatomite to Prevent Precipitate Adhesion
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Solution Overview
Problem
Existing methods for purifying factor XIII face challenges in achieving high yield and purity due to precipitate adherence to impellers during salt precipitation, which complicates large-scale production.
Innovation Solution
A method involving diatomite and salt precipitation steps, followed by heat treatment and additional salt precipitation, along with solvent/detergent treatment and anion exchange chromatography, to enhance purification efficiency and prevent precipitate adherence.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional salt precipitation is used to purify factor XIII, then purification is achieved, but precipitate adheres to impeller causing low yield and operational difficulties
Solution Approach 1:
Diatomite is introduced as an intermediary substance during the salt precipitation process. The diatomite acts as a nucleation site and carrier for the precipitating factor XIII, preventing direct adhesion to the impeller while maintaining effective precipitation. This mediator enables the precipitate to be easily suspended and later removed without equipment fouling.
Solution Approach 2:
The invention modifies the physical parameters of the precipitation process by controlling temperature (4-37°C), salt concentration (20-40% w/v), and diatomite dosage (0.1-10% w/v). These parameter optimizations ensure complete precipitation of factor XIII while maintaining precipitate suspension and preventing impeller adhesion.
2Manufacturing precision
If multiple purification steps are added to increase purity, then purification quality improves, but process complexity and cost increase
Solution Approach 1:
The invention combines multiple functions into the salt precipitation step: factor XIII precipitation, impeller protection via diatomite, and preliminary concentration. This integrated approach achieves high purity (80%) without requiring separate complex purification stages, simplifying the overall process while maintaining effectiveness.
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 method achieves high yield and purity of factor XIII, suitable for large-scale industrial production, with a purity of about 80% compared to conventional methods' 50%, and is economically viable.
Implementation Method 1
precipitating by adding diatomite and a salt to a separated plasma sample containing factor XIII
Implementation Method 2
precipitating by adding diatomite and a salt to a separated plasma sample containing factor XIII
Implementation Method 3
obtaining a solution by dissolving the precipitate obtained by first salt precipitation in step (a) in a dissolution buffer, and then treating heat (heat treatment)
Implementation Method 4
precipitating by adding a salt to the solution heat-treated in step (b)
Data Source
AI summary
A method of purifying factor XIII (FXIII), including performing a first salt precipitation by adding diatomite and a salt to a separated plasma sample containing factor XIII, and then obtaining a precipitate by removing a supernatant, dissolving the precipitate in a dissolution buffer, and then treating with heat to form a solution, and performing a second salt precipitation by adding a salt to the solution and then obtaining a precipitate by removing a supernatant.