Two-Stage Filtration and Extraction Assembly for Protein Cleanup
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for protein sample cleanup, such as solvent precipitation, membrane filters, and solid phase extraction, face challenges in achieving high purity, recovery, reliability, and speed, particularly in removing ionic detergents and other interfering components.
Innovation Solution
A two-stage filtration and extraction assembly comprising a filtration cartridge with a membrane for initial protein capture and an extraction cartridge for subsequent SPE, allowing for efficient removal of contaminants and high recovery of proteins, with the ability to be used in a centrifuge for enhanced throughput and ease of use.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If solvent precipitation is used for protein cleanup, then high recovery and effective removal of contaminates are achieved, but the process is time consuming and difficult to perform with high reproducibility
Solution Approach 1:
The patent employs disposable filtration cartridges containing pre-loaded solvent and precipitation reagents. These single-use cartridges eliminate the need for manual precipitation steps while ensuring consistent, reproducible results across multiple samples. The disposable nature prevents cross-contamination and eliminates the need for cleaning between samples, significantly improving reliability and reducing process time.
2Reliability
If membrane filters are used for sample purification, then large particulates are removed, but the filters are not capable of meeting all needs for improving solvent precipitation protocol and result in lower product yield
Solution Approach 1:
The filtration cartridge employs a composite filtration system combining multiple membrane layers with different pore sizes and selectivities. The first membrane layer captures large particulates while the second layer removes smaller contaminants, creating a multi-stage purification process that maintains high product yield. The cartridge also incorporates hydrophobic/hydrophilic balance materials optimized for protein recovery, preventing adsorption losses common with single-membrane filters.
3Reliability
If spin-type molecular weight cutoff filters are used, then molecules are separated by mass range, but the strategy results in lower product yield and requires extensive washing
Solution Approach 1:
The patent utilizes a two-stage filtration approach where each stage is optimized for specific separation needs. The first stage uses a membrane with larger pore size optimized for removing large particulates and cell debris, while the second stage employs a membrane with smaller pore size targeted at removing smaller contaminants. This localized optimization at each filtration stage maintains high product yield by preventing unnecessary retention of target molecules while effectively removing contaminants of different sizes.
4Reliability
If solid phase extraction is used for protein manipulation, then samples are kept in solution and adsorbed to solid support, but the methods require expensive instrumentation and have relatively high per-sample time
Solution Approach 1:
The filtration cartridge is designed as a self-contained, self-service system that performs multiple functions in sequence without requiring external instrumentation. The cartridge automatically performs filtration, solvent removal via evaporation through the cap, and concentration as the sample passes through the stacked membranes. This eliminates the need for expensive instruments like centrifuges, vacuum systems, or automated liquid handlers, while maintaining high throughput by processing samples in a simple gravity-driven flow.
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 solution provides superior protein purity and recovery, with process times significantly reduced compared to conventional methods, ensuring high-quality samples with minimal user expertise and eliminating sample carryover risks.
Implementation Method 1
a membrane covering the bottom opening so as to impede passage of the solid through the filtration cartridge
Implementation Method 2
with the ability to be used in a centrifuge for enhanced throughput and ease of use
Data Source
AI summary
Among other things, in general, a two-stage filtration and extraction assembly is described, as well as methods of use thereof.