Membrane Extraction Filter Cartridge for Rapid Cell Disruption
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Solution Overview
Problem
Current methods for isolating membrane and organelle proteins are time-consuming, require large sample quantities, and often alter the native configuration of proteins, with traditional protocols involving glass homogenizers or blenders being cumbersome and prone to cross-contamination.
Innovation Solution
The use of membrane extraction filter cartridges with hydrophobic porous materials and defined pore sizes for rapid disruption of cells, allowing for the separation of retentate and filtrate under centrifugation or vacuum, which retains cellular debris while releasing ruptured membranes and associated proteins, enabling native form isolation in under 30 minutes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional glass homogenizers or blenders are used for cell disruption, then membrane proteins can be isolated in their native forms, but the procedure becomes time-consuming and cumbersome requiring thorough cleaning between samples
Solution Approach 1:
The patent extracts the cell disruption function from traditional glass homogenizers and blenders by using pre-packaged filter cartridges containing rigid support beads. This extraction eliminates the need for cleaning complex homogenizing devices between samples while maintaining effective cell disruption through the bead-mediated mechanical stress during filtration
Solution Approach 2:
The patent employs disposable filter cartridges containing rigid support beads instead of reusable glass homogenizers. These single-use cartridges are discarded after one sample processing, eliminating time-consuming cleaning procedures while providing consistent cell disruption performance across multiple samples
2Quantity of substance
If detergents are used for membrane protein isolation, then smaller amounts of starting cells are required and selective extraction is achieved, but the normal configuration of membrane proteins is potentially altered
Solution Approach 1:
The patent changes the physical parameter of cell disruption by using mechanical stress through rigid support beads during filtration, replacing the chemical parameter approach of detergent-based lysis. This physical disruption method achieves effective cell breaking with minimal starting material while preserving protein native conformation by avoiding chemical denaturation
3Reliability
If multiple incubation and centrifugation steps are performed as in current commercial kits, then membrane proteins are isolated, but the procedure becomes tedious and time-consuming taking more than one hour
Solution Approach 1:
The patent merges cell disruption, filtration, and initial separation steps into a single integrated operation using rigid support beads within the filter cartridge. This consolidation combines mechanical disruption and filtration that were traditionally separate steps, achieving complete membrane protein isolation in under 30 minutes while maintaining isolation completeness
4Reliability
If glass homogenizers are used for multiple samples, then membrane proteins can be isolated, but thorough cleaning is required for each sample creating possibility of cross-contamination
Solution Approach 1:
The patent replaces reusable glass homogenizers with disposable filter cartridges containing rigid support beads. Each cartridge is used for a single sample and then discarded, eliminating cross-contamination risks entirely while maintaining effective membrane protein isolation. This disposable approach significantly improves user friendliness by removing complex cleaning procedures
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 significantly reduces isolation time, maintains protein native conformation, and is user-friendly for both single and multiple samples, providing high yield and consistent results, making it the fastest and most efficient method for membrane and organelle isolation.
Implementation Method 1
the filter is a surface filter (e.g., a filter that traps certain cellular components on its surface)... Upon application of positive (such as centrifugation) or negative force (such as vacuum suction), the mixture of biological samples are forced to pass through the filter, resulting in two separated parts: retentate (on the surface and inside of the filter), and filtrate (passing through the filter)
Implementation Method 2
Upon application of positive (such as centrifugation) or negative force (such as vacuum suction), the mixture of biological samples are forced to pass through the filter
Data Source
AI summary
Disclosed herein are methods for quickly obtaining crude membranes from cells, including animal and plant cells. The methods include incubating cells in a buffer and forcing the cells through a filter that causes rupture of the cells, and then separating the resulting crude membranes from most cytosolic proteins.