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

Andrew R Hanna1, Sarah J Shepherd1, Gregory A Datto1

  • 1Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA 19104.

Biorxiv : the Preprint Server for Biology
|June 12, 2025
PubMed
Summary

This study introduces a high-throughput microfluidic platform for rapid lipid nanoparticle (LNP) formulation. The automated system accelerates LNP library generation for therapeutic applications.

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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.