Robust, Scalable Microfluidic Manufacturing of RNA-Lipid Nanoparticles Using Immobilized Antifouling Lubricant...

Area of Science:

• Biotechnology
• Materials Science
• Chemical Engineering

Background:

• Microfluidic mixing offers advantages for RNA-loaded lipid nanoparticle (RNA-LNP) production, including precise size control and high encapsulation efficiency.
• Commercial translation of microfluidic RNA-LNP production is hindered by device surface fouling during prolonged operation.
• The complex composition of RNA-LNPs makes finding effective antifouling coatings challenging.

Purpose of the Study:

• To develop an effective antifouling strategy for microfluidic devices used in RNA-LNP production.
• To enhance the stability and reliability of microfluidic systems for scalable RNA-LNP manufacturing.
• To demonstrate the compatibility of the antifouling approach with commercial-scale production.

Main Methods:

• An immobilized liquid lubricant layer of perfluorodecalin (PFD) was applied to a staggered herringbone microfluidic (SHM) mixing chip.
• The antifouling performance was evaluated over extended operational periods.
• The method was tested on a parallelized microfluidic platform with 256 SHM mixers for scalability.
• Produced RNA-LNPs were characterized for physiochemical properties and validated for in vitro and in vivo mRNA delivery.

Main Results:

• The PFD lubricant layer enabled over 3 hours of stable microfluidic operation, a >15-fold increase compared to standard methods.
• The antifouling technology was successfully integrated into a 256-mixer parallelized platform, demonstrating scalable production at L/h rates.
• RNA-LNPs produced using the antifouling method exhibited comparable physiochemical properties and mRNA delivery performance to those made without the coating.

Conclusions:

• Immobilized liquid lubricant layers effectively suppress surface fouling in microfluidic devices for RNA-LNP production.
• This antifouling strategy significantly enhances the reliability and operational stability of microfluidic manufacturing processes.
• The technology holds promise for transforming the microfluidic production of diverse materials, enabling robust and scalable manufacturing.