LNP Filtration Using Single-Pass TFF to Limit Shear Damage
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Solution Overview
Problem
Traditional methods for producing lipid nanoparticles (LNPs) for delivering biologically active substances like nucleic acids face challenges in maintaining potency and reducing toxicity due to hydrodynamic shear forces during recirculation in tangential flow filtration, which affects the effective delivery of nucleic acids to cells.
Innovation Solution
A method involving single-pass tangential flow filtration (SPTFF) is employed to produce LNPs, where nucleic acids and ionizable lipids are introduced into a reactor at different flow rates, followed by filtering the reaction mixture without recirculation, and using a sterile filtration apparatus to produce a retentate, enhancing the encapsulation efficiency and reducing toxicity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional tangential flow filtration with recirculation is used, then the desired concentration and buffer composition are achieved, but the LNP potency decreases due to hydrodynamic shear forces
Solution Approach 1:
The patent applies single-pass tangential flow filtration instead of recirculation, allowing the LNP to pass through the filtration system once at controlled conditions to achieve buffer exchange and concentration without repeated exposure to hydrodynamic shear forces that would damage the nanoparticles and reduce potency
Solution Approach 2:
The patent modifies the filtration parameters by controlling the feed flux within a specific range (10-200 L/m2/hour) and using a single-pass approach rather than recirculation, changing the physical conditions to minimize shear stress on LNPs while still achieving the required concentration and buffer composition
2Manufacturing precision
If recirculation is used in tangential flow filtration, then the processing time is extended, but the LNP potency is adversely impacted
Solution Approach 1:
The patent implements a single-pass filtration approach that completes concentration and buffer exchange in one continuous operation, eliminating the need for multiple recirculation cycles and thereby reducing processing time while still achieving the desired LNP concentration
3Reliability
If single-pass tangential flow filtration is used, then the exposure to hydrodynamic shear forces is minimized, but the encapsulation efficiency must be maintained
Solution Approach 1:
The patent optimizes the feed flux parameter within a specific range (10-200 L/m2/hour) to balance two competing requirements: minimizing hydrodynamic shear forces that would reduce encapsulation efficiency while still achieving effective buffer exchange and concentration in a single pass, thereby reducing toxicity without compromising encapsulation
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 SPTFF method improves the encapsulation efficiency and reduces toxicity of LNPs, ensuring effective delivery of nucleic acids by minimizing exposure to hydrodynamic shear forces, thereby enhancing the therapeutic potential of LNPs.
Implementation Method 1
filtering the nanoparticle from the reaction mixture via a single-pass tangential flow filtration system at a feed flux to provide a retentate
Implementation Method 2
filtering the retentate through a sterile filtration apparatus before or after filtering the nanoparticle from the reaction mixture
Data Source
AI summary
A method is provided for producing a composition that includes a nanoparticle with at least one nucleic acid and at least one ionizable lipid. The method includes introducing a first composition that includes the at least one nucleic acid and a second composition that includes the at least one ionizable lipid into at least one reactor. The first composition and the second composition are introduced at a first flow rate and a second flow rate, respectively, therefore generating the nanoparticle in a reaction mixture. The method further includes filtering the nanoparticle from the reaction mixture via a single-pass tangential flow filter at a feed flux to provide a retentate, to produce the composition. The method may also include filtering the retentate through a sterile filtration membrane.