A preparation method for mRNA-LNPs with improved properties

Area of Science:

• Biotechnology
• Nanomedicine
• Drug Delivery

Background:

• Messenger RNA lipid nanoparticles (mRNA-LNPs) are crucial for various therapeutic applications.
• The efficacy and safety of mRNA-LNPs are significantly influenced by their physicochemical properties.
• Preparation methods directly impact critical parameters such as size, purity, stability, and potency.

Purpose of the Study:

• To introduce and evaluate a novel Two-step tangential-flow filtration (TFF) method for mRNA-LNP preparation.
• To investigate the impact of this improved method on key mRNA-LNP characteristics.
• To assess the potential of the optimized mRNA-LNPs for industrial applicability and clinical use.

Main Methods:

• Implementation of a Two-step tangential-flow filtration (TFF) process for mRNA-LNP production.
• Inclusion of an additional ethanol removal step before particle fusion.
• Characterization of mRNA-LNPs for size, empty particle content, morphology, storage stability, and in vitro transfection efficiency.
• Evaluation of in vivo biodistribution, particularly in the heart and blood.

Main Results:

• The Two-step TFF method successfully produced larger mRNA-LNPs.
• A significant reduction in the proportion of empty LNPs was observed.
• Optimized storage stability was achieved, maintaining integrity for at least 6 months at 2-8 °C.
• Enhanced in vitro transfection efficiency and improved in vivo biodistribution with minimized cardiac and blood accumulation were demonstrated.

Conclusions:

• The Two-step TFF method offers a significant advancement in mRNA-LNP preparation.
• This method enhances critical mRNA-LNP properties, including purity, stability, and efficacy.
• The optimized mRNA-LNPs show promise for improved industrial scalability and expanded clinical applications.