Automated and parallelized microfluidic generation of large and precisely-defined lipid nanoparticle libraries

Area of Science:

• Biotechnology
• Materials Science
• Nanotechnology

Background:

• Lipid nanoparticles (LNPs) show promise beyond vaccines for various therapeutic applications.
• Current LNP discovery is limited by slow, manual microfluidic formulation processes.
• High-throughput methods are needed to accelerate LNP development.

Purpose of the Study:

• To develop and validate a high-throughput, automated microfluidic platform for parallel LNP library generation.
• To overcome the rate-limiting step in LNP formulation for drug discovery.
• To enable rapid physicochemical and biological profiling of LNP libraries.

Main Methods:

• An automated microfluidic platform with eight parallel mixers was designed.
• Lithographically encoded fluidic resistors and controlled pressure supplies varied reagent flow ratios.
• Custom frobotic plate handling enabled rapid collection of distinct LNP formulations.
• A library of 96 unique LNP formulations was generated.

Main Results:

• The platform achieved a formulation rate of one distinct LNP every three seconds.
• Physicochemical profiling of the 96-formulation library was performed.
• In vitro and in vivo transfection efficiencies of the generated LNPs were evaluated.

Conclusions:

• The developed automated microfluidic platform significantly accelerates LNP library production.
• This high-throughput approach facilitates rapid discovery and optimization of LNPs for therapeutic applications.
• The platform enables efficient screening of LNP formulations for enhanced drug delivery.