Pressurized Nucleic Acid Filtration for Low-Loss Buffer Replacement
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Solution Overview
Problem
Existing methods for processing nucleic acids, such as dialysis and centrifuge-based ultrafiltration, are labor-intensive, time-consuming, and result in significant sample loss, especially when dealing with small sample volumes.
Innovation Solution
A method involving a filter housing that concentrates nucleic acids by pressurization and depressurization, using a pressure gradient to separate nucleic acids from contaminants, with automated cycles to maintain molecular integrity and achieve complete buffer replacement.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional dialysis is used for nucleic acid processing, then contaminants can be removed, but the process requires large sample volumes, takes several hours, and results in significant sample loss
Solution Approach 1:
The patent replaces the mechanical dialysis process with a pressure-driven filtration system using a syringe and filter unit. By applying mechanical pressure to force the liquid through a semipermeable membrane, the process achieves contaminant removal in seconds rather than hours, directly resolving the time efficiency contradiction while maintaining purification effectiveness
Solution Approach 2:
The patent employs a semipermeable membrane with specific pore sizes that allow contaminants to pass through while retaining nucleic acids. This porous material enables rapid separation based on molecular size differences, achieving both efficient contaminant removal and significantly reduced processing time compared to conventional dialysis
2Productivity
If centrifuge-based ultrafiltration is used, then nucleic acids can be concentrated, but the method is labor intensive and time-consuming
Solution Approach 1:
The patent designs a self-contained filter unit where the nucleic acid suspension is directly loaded into the syringe barrel and processed in-place. The system performs filtration and concentration automatically through pressure application without requiring external centrifugation equipment or multiple transfer steps, thereby simplifying operation while maintaining high productivity
Solution Approach 2:
The patent combines the filtration chamber, membrane, and collection system into a single integrated filter unit. By merging these components, the system eliminates the need for separate centrifugation and transfer steps, reducing operational complexity while achieving efficient nucleic acid concentration in one continuous process
3Quantity of substance
If small sample volumes are processed by conventional methods, then sample loss becomes even more pronounced
Solution Approach 1:
The patent extracts only the necessary filtration function from complex processing systems and implements it through a simple syringe-based filter unit. This minimalistic approach processes small sample volumes directly without requiring large buffer volumes for dialysis or multiple centrifugation steps, thereby minimizing sample loss while effectively concentrating the nucleic acid
Solution Approach 2:
The patent performs filtration and concentration in a single preliminary step using the filter unit, rather than requiring multiple sequential processing steps. By achieving contaminant removal and concentration in one operation, the system minimizes handling and transfer steps that would otherwise cause sample loss, particularly critical when working with small sample volumes
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 efficiently concentrates nucleic acids, reduces sample loss, and automates the process, providing a rapid and effective solution for nucleic acid processing.
Implementation Method 1
a filter that does not pass the nucleic acid
Implementation Method 2
pressurizing the housing to produce a filtrate and a nucleic acid retentate from the initial suspension
Implementation Method 3
increasing the pressure of the housing by injecting a gas (e.g., air, argon, N2, or CO2) into the housing
Data Source
AI summary
The invention features methods and apparatus for concentrating a nucleic acid. Methods of the invention include providing an initial suspension of a nucleic acid and an initial liquid, contacting the initial suspension with a housing having a filter that does not pass the nucleic acid, pressurizing the housing to produce a filtrate and a nucleic acid retentate from the initial solution, and detecting the volume of the nucleic acid retentate. Apparatus of the invention include a chamber configured to hold a filter housing containing a nucleic acid suspension, a pressure source to filter the suspension, and a detector to depressurize the housing upon detecting the volume reaching a predetermined threshold. The methods and apparatus described herein are useful in filtering, concentrating, and reconstituting nucleic acids, such as mRNA, in processes such as complete buffer replacement.