Normal Flow Filtration for mRNA Purification
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Solution Overview
Problem
Current methods for purifying mRNA synthesized in vitro are limited in scale and unable to produce high-quality, clinical-grade mRNA due to contamination issues, restricting their use in therapeutic applications.
Innovation Solution
A method utilizing normal flow filtration to purify mRNA, where the mRNA is precipitated, retained on a filter, washed, and then dissolved to remove contaminants, allowing for high-yield, high-purity mRNA production suitable for therapeutic use without the need for additional purification steps.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If traditional silica-based column systems or organic extraction methods are used for mRNA purification, then small scale purification (5-10 mg) is achieved, but the scale is insufficient for clinical and commercial therapeutic uses
Solution Approach 1:
The invention changes the physical parameters of the filtration process by using normal flow filtration with specific pore size filters (0.22 µm or 0.45 µm) instead of traditional column chromatography parameters, enabling scale-up from 5-10 mg to gram-scale purification while maintaining purity and yield
Solution Approach 2:
The invention employs porous filter membranes with defined pore sizes (0.22 µm or 0.45 µm) to physically separate mRNA from contaminants through size exclusion, replacing the silica-based adsorption mechanism and enabling larger scale processing
2Quantity of substance
If traditional purification methods are used, then small scale production is achieved, but contamination with proteins, salts, buffers, and non-RNA nucleic acids remains problematic
Solution Approach 1:
The invention extracts and removes contaminants (proteins, salts, buffers, non-RNA nucleic acids) from the mRNA preparation through normal flow filtration, achieving clinical-grade purity (>95%) while recovering high yields of intact mRNA
Solution Approach 2:
The invention changes the purification mechanism from chemical adsorption (silica columns) or organic extraction to physical filtration based on size exclusion, achieving both high purity and high yield simultaneously at clinical scales
3Reliability
If in vitro transcription systems are used to synthesize mRNA, then full-length mRNA is produced, but the reaction composition contains undesirable contaminants that must be removed
Solution Approach 1:
The invention converts the challenge of removing diverse synthesis contaminants into a benefit by using normal flow filtration that simultaneously removes proteins, salts, buffers, and non-RNA nucleic acids in a single step, achieving clinical-grade purity while preserving mRNA integrity
Solution Approach 2:
The invention changes the approach from multiple sequential purification steps to a single normal flow filtration step that removes all types of contaminants generated during in vitro transcription, simplifying the process while maintaining high mRNA integrity
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 achieves high recovery and integrity of mRNA, with up to 99% yield and >95% purity, meeting clinical-grade standards without further purification, enabling large-scale, cost-effective production of high-quality mRNA for therapeutic applications.
Implementation Method 1
subjecting the suspension comprising the precipitated mRNA to a filter wherein the precipitated mRNA is retained by the filter
Implementation Method 2
the precipitated mRNA is retained by the filter
Data Source
AI summary
The present invention provides, among other things, methods for purifying mRNA based on normal flow filtration for therapeutic use.