Recombinant Protein Aggregate Separation via Metal Salt Flocculation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods are inefficient in separating insoluble recombinant protein aggregates from recombinant cells, particularly when the proteins are in a solubilized state or form fine insoluble granules that are difficult to centrifuge.
Innovation Solution
A method involving the disruption of recombinant cells, aggregation of insoluble bodies using metal salts, acids, or anionic flocculants, followed by centrifugation or filtration to produce a recombinant protein aggregate with a particle size of 4 μm to 50 μm, utilizing centrifugal forces of 10,000×g or less and specific separation techniques.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If centrifugation is used to separate insoluble granules, then separation can be achieved, but fine insoluble granules are difficult to separate efficiently
Solution Approach 1:
The patent changes the physical-chemical parameters of the insoluble granules by adding metal salts, acids, or anionic flocculants to induce aggregation. This transforms fine granules into larger aggregates with different settling characteristics, enabling efficient separation through low-speed centrifugation or filtration.
Solution Approach 2:
The patent introduces intermediary substances (metal salts, acids, anionic flocculants) that mediate between the fine insoluble granules and the separation process. These intermediaries facilitate aggregation of granules into larger structures that are easier to separate using conventional low-speed centrifugation or filtration methods.
2Productivity
If high-speed centrifugation is used to separate fine insoluble granules, then separation can be achieved, but energy consumption and equipment requirements increase
Solution Approach 1:
The patent modifies the physical parameters of the suspended particles through aggregation induced by metal salts, acids, or anionic flocculants. This changes the settling velocity and allows separation using low-speed centrifugation (1,000-10,000×g) instead of high-speed centrifugation, significantly reducing energy consumption.
Solution Approach 2:
The patent replaces the mechanical system of high-speed centrifugation with a combination of chemical aggregation and low-speed centrifugation or filtration. This substitution reduces the mechanical energy input required while achieving effective separation.
3Productivity
If recombinant protein is produced as insoluble granules, then high yield and purity can be achieved, but control over granule formation is difficult
Solution Approach 1:
The patent systematically varies physical and chemical parameters (temperature, pH, metal ion concentration, flocculant type and concentration) to control and optimize granule formation. This allows adjustment of granule size, density, and formation kinetics to achieve desired yield and purity.
Solution Approach 2:
The patent employs dynamic control of aggregation conditions, allowing the system to transition from controlled granule formation to efficient separation. The aggregation process is made reversible and controllable through pH changes, metal ion addition, or flocculant introduction, enabling optimization at each stage.
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 method enables efficient separation and purification of recombinant protein aggregates, improving their yield and purity, and facilitating industrial-scale production by overcoming the challenges of centrifuging fine insoluble granules.
Implementation Method 1
aggregating the insoluble bodies of the recombinant protein
Implementation Method 2
anionic flocculants, followed by centrifugation or filtration
Implementation Method 3
separating the resulting aggregate
Implementation Method 4
centrifugation or filtration to produce a recombinant protein aggregate
Data Source
AI summary
An object of the present invention is to provide a method for efficiently separating insoluble bodies of a recombinant protein from a recombinant cell expressing a target recombinant protein as insoluble bodies in the cell. The present invention provides a method for producing a recombinant protein aggregate by separating insoluble bodies of a recombinant protein from a recombinant cell expressing the recombinant protein as insoluble bodies in the cell, including disrupting the recombinant cell, aggregating the insoluble bodies of the recombinant protein, and separating the resulting aggregate.