Automated and Parallelized Microfluidic Generation of Large and Precisely Defined Lipid Nanoparticle Libraries.

Area of Science:

• Biotechnology
• Materials Science
• Nanotechnology

Background:

• Lipid nanoparticles (LNPs) are crucial for therapeutic applications, but their formulation is a bottleneck in drug discovery.
• Current methods for LNP formulation are manual and sequential, limiting throughput.
• Advancements in lipid synthesis and screening necessitate faster formulation methods.

Purpose of the Study:

• To develop a parallelized, automated microfluidic platform for high-throughput LNP library generation.
• To overcome the rate-limiting step in early-stage LNP discovery and development.
• To bridge the gap between LNP discovery and manufacturing.

Main Methods:

• A parallelized microfluidic platform with eight microscale mixers was designed.
• Lithographically encoded fluidic resistors and controlled pressure supplies enabled precise formulation control.
• Custom robotic plate handling was integrated for rapid collection of distinct LNP formulations.
• On-chip characterization of physicochemical properties and in vitro transfection efficiency was performed.

Main Results:

• The platform achieved a throughput of approximately 1000 distinct LNP formulations per hour.
• 96 formulations were characterized, demonstrating precise control over physicochemical properties and transfection efficiency.
• A lead candidate formulation was identified and validated in vivo.
• The platform demonstrated scalability from discovery to liters per hour production.

Conclusions:

• The automated microfluidic platform significantly accelerates LNP formulation and discovery.
• This technology enables rapid identification and optimization of LNP candidates for therapeutic use.
• The parallelization approach bridges the gap between early-stage research and large-scale manufacturing.