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Formulating and Characterizing Lipid Nanoparticles for Gene Delivery using a Microfluidic Mixing Platform
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High-Throughput Formulation Using Fast mRNA Lipid Nanoparticle Assembly via Simple Thermomixing (HTF-FLASH).

Soyeon Yoo1,2, Emily L Han1, Amanda M Murray1

  • 1Department of Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.

Nano Letters
|September 25, 2025
PubMed
Summary
This summary is machine-generated.

High-throughput formulation (HTF) using the fast lipid nanoparticle assembly via simple thermomixing (HTF-FLASH) technique enables rapid and uniform production of lipid nanoparticles (LNPs). This method accelerates screening workflows, offering an efficient alternative to traditional LNP formulation strategies.

Keywords:
formulation methodologygene deliveryhigh-throughput formulationlipid nanoparticlesnucleic acid

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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Drug Delivery

Background:

  • Ionizable lipid nanoparticles (LNPs) are crucial for nucleic acid delivery.
  • High-throughput formulation (HTF) is essential for identifying optimal LNPs.
  • Conventional methods like manual pipetting and microfluidics present challenges for HTF.

Purpose of the Study:

  • To introduce a novel HTF method for LNP formulation.
  • To evaluate the efficiency and reproducibility of the new method.
  • To compare HTF-FLASH with existing LNP formulation techniques.

Main Methods:

  • Development of the fast lipid nanoparticle assembly via simple thermomixing (HTF-FLASH) technique.
  • Utilizing a thermomixer and 96-deep-well plates for parallel LNP production.
  • Assessing physicochemical properties and mRNA transfection efficiencies (in vitro and in vivo).

Main Results:

  • HTF-FLASH enables rapid and uniform LNP production within a minute.
  • Formulated LNPs demonstrated comparable physicochemical properties to microfluidic-formulated LNPs.
  • In vitro and in vivo mRNA transfection efficiencies were similar to those achieved with microfluidics.

Conclusions:

  • HTF-FLASH is a high-throughput, efficient, and reproducible method for LNP screening.
  • This technique offers a cost-effective alternative to traditional LNP formulation.
  • Accelerates the discovery and development of nucleic acid delivery systems.