Compressible Foam Filter for Lysate Viscosity Reduction
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Solution Overview
Problem
Current methods for reducing the viscosity of cell or tissue lysates often require specialized equipment, are time-consuming, or result in protein denaturation, making them inefficient for downstream analysis and purification processes.
Innovation Solution
A method involving a compressible and open-cell foam filter with specific pore sizes and compression ratios is used to separate genomic DNA from polypeptides in a lysate, allowing for efficient viscosity reduction without the need for specialized equipment and minimizing protein loss or denaturation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-affected harmful factors
If sonication is used to break up genomic DNA and reduce viscosity, then viscosity is reduced, but target protein may be partially or totally denatured
Solution Approach 1:
The patent removes genomic DNA from the lysate through filtration using a syringe filter, extracting the harmful component (DNA causing viscosity) without applying mechanical stress to the proteins. This separates the DNA removal function from protein-stressing methods like sonication.
Solution Approach 2:
The patent replaces mechanical breakdown methods (sonication, needle passage) with a chemical/biological approach using DNase I enzyme to digest genomic DNA, followed by gentle filtration. This substitutes harsh mechanical energy with enzymatic action and low-stress filtering.
2Object-affected harmful factors
If passage through fine gauge needle is used to reduce viscosity, then viscosity is reduced, but the method is very time consuming and tedious and does not scale well
Solution Approach 1:
The patent divides the DNA removal process into distinct functional steps: enzymatic digestion phase followed by filtration phase. This segmentation allows each step to be optimized independently and enables parallel processing of multiple samples through the filtration system.
Solution Approach 2:
The syringe filter system serves multiple functions: it acts as both the filtration medium and the collection device, and can process various lysate volumes by simply changing the syringe size. This universal design eliminates the need for sample-specific optimization required by needle passage methods.
3Object-affected harmful factors
If compaction agent such as spermidine is used to reduce viscosity, then viscosity is reduced, but additional reagents and steps are required
Solution Approach 1:
The patent combines the DNA digestion and filtration steps into a continuous process where the lysate flows from the DNase I reaction directly through the syringe filter. This merging eliminates intermediate transfer steps and reduces the number of separate reagents needed compared to compaction agent methods.
Solution Approach 2:
The syringe filter system is self-contained and requires no external equipment or additional reagents beyond what is already in the lysis buffer. The filter membrane itself performs the separation function without needing chemical aids, making the process simpler than compaction agent approaches.
4Quantity of substance
If specialized equipment such as centrifuge or vacuum is used to filter lysate, then filtration is effective, but the equipment requirement limits usefulness
Solution Approach 1:
The syringe filter system uses the natural pressure of the liquid flow and the mechanical action of drawing/plunger movement to drive filtration. No external vacuum or centrifugal force is needed, making the system portable and usable in any laboratory setting without specialized equipment.
Solution Approach 2:
The patent replaces complex mechanical separation systems (centrifuges, vacuum filtration apparatus) with a simple manual syringe filter system. The filtration is driven by manual plunger movement rather than external mechanical forces, dramatically simplifying the equipment requirements while maintaining effective separation.
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 effectively reduces lysate viscosity by retaining genomic DNA while preserving over 95% of the original polypeptides, enabling more efficient downstream processing such as Western blot analysis and protein purification without the use of centrifuges or vacuums.
Implementation Method 1
contacting the cell lysate or tissue lysate with a compressible and open-cell foam filter having a pore size from about 0.65 mm to about 1.22 mm
Implementation Method 2
compressing the filter to recover the lysate absorbed in the filter, wherein the compression ratio is from about 50% to about 95%
Data Source
AI summary
The present disclosure provides methods for reducing the viscosity of a cell lysate or tissue lysate by: contacting the cell lysate or tissue lysate with a compressible and open-cell foam filter having a pore size from about 0.65 mm to about 1.22 mm, compressing the filter to recover the lysate absorbed in the filter, and collecting the filtered lysate; and also provides kits therefor.