2. Academic Validation
  3. Robust, Scalable Microfluidic Manufacturing of RNA-Lipid Nanoparticles Using Immobilized Antifouling Lubricant Coating

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

  • ACS Nano. 2025 Jan 14;19(1):1090-1102. doi: 10.1021/acsnano.4c12965.
Yoon-Ho Hwang 1 2,  Sarah J Shepherd 3,  Dongyoon Kim 3,  Alvin J Mukalel 3,  Michael J Mitchell 3 4 5 6 7 8 9 10,  David A Issadore 1 3 10,  Daeyeon Lee 1
Affiliations

Affiliations

  • 1 Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • 2 Department of Polymer Engineering, Pukyong National University, Nam-gu, Busan 48513, Republic of Korea.
  • 3 Department of Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • 4 Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • 5 Institute for Immunology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • 6 Cardiovascular Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • 7 Institute for Regenerative Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • 8 Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • 9 Penn Institute for RNA Innovation, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • 10 Department of Electrical and Systems Engineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
PMID: 39700475 DOI: 10.1021/acsnano.4c12965
Abstract

Despite the numerous advantages demonstrated by microfluidic mixing for RNA-loaded lipid nanoparticle (RNA-LNP) production over bulk methods, such as precise size control, homogeneous distributions, higher encapsulation efficiencies, and improved reproducibility, their translation from research to commercial manufacturing remains elusive. A persistent challenge hindering the adoption of microfluidics for LNP production is the fouling of device surfaces during prolonged operation, which significantly diminishes performance and reliability. The complexity of LNP constituents, including lipids, Cholesterol, RNA, and solvent mixtures, makes it difficult to find a single coating that can prevent fouling. To address this challenge, we propose using an immobilized liquid lubricant layer of perfluorodecalin (PFD) to create an antifouling surface that can repel the multiple LNP constituents. We apply this technology to a staggered herringbone microfluidic (SHM) mixing chip and achieve >3 h of stable operation, a >15× increase relative to gold standard approaches. We also demonstrate the compatibility of this approach with a parallelized microfluidic platform that incorporates 256 SHM mixers, with which we demonstrate scale up, stable production at L/h production rates suitable for commercial scale applications. We verify that the LNPs produced on our chip match both the physiochemical properties and performance for both in vitro and in vivo mRNA delivery as those made on chips without the coating. By suppressing surface fouling with an immobilized liquid lubricant layer, this technology not only enhances RNA-LNP production but also promises to transform the microfluidic manufacturing of diverse Materials, ensuring more reliable and robust processes.

Keywords

nanoprecipitation; omniphobic coating; parallelization; staggered herringbone microfluidic mixer; vaccine.

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