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Process Optimization of Charge-Reversible Lipid Nanoparticles for Cytosolic Protein Delivery Using the

Dai Oyama1,2, Masako Okada1, Furan Song1

  • 1Laboratory of Medical Biochemistry, University of Shizuoka School of Pharmaceutical Sciences, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.

Biological & Pharmaceutical Bulletin
|March 25, 2025
PubMed
Summary

Researchers optimized manufacturing parameters for dioleoylglycerophosphate-diethylenediamine (DOP-DEDA)-based lipid nanoparticles (LNPs) for protein delivery. This process yields high-quality, pH-responsive LNPs with controlled particle size and encapsulation efficiency for intracellular applications.

Keywords:
Box–Behnken designcytosolic protein deliverydesign of experimentdioleoylglycerophosphate–diethylenediaminelipid nanoparticleprocess optimization

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Area of Science:

  • Biotechnology
  • Materials Science
  • Nanotechnology

Background:

  • Lipid nanoparticles (LNPs) are crucial for drug delivery.
  • Dioleoylglycerophosphate-diethylenediamine (DOP-DEDA) is a pH-responsive lipid enabling intracellular cargo delivery.
  • Optimizing LNP manufacturing is key for effective protein therapeutics.

Purpose of the Study:

  • To determine optimal manufacturing parameters for protein-encapsulated DOP-DEDA-based LNPs.
  • To achieve desired quality characteristics, including particle size and encapsulation efficiency.
  • To establish a reproducible process for intracellular protein drug delivery.

Main Methods:

  • Utilized microfluidic technology for LNP preparation.
  • Employed Box-Behnken design for process optimization.
  • Evaluated particle size, ζ-potential, and encapsulation efficiency against process variables.

Main Results:

  • Lipid solution ratio and total flow rate significantly impacted particle size and encapsulation efficiency, respectively.
  • DOP-DEDA LNPs demonstrated pH-responsive charge reversibility.
  • Achieved reproducible 100-nm LNPs with 27% encapsulation efficiency within predicted ranges.

Conclusions:

  • Established manufacturing process parameters for optimal protein-encapsulated DOP-DEDA-based LNPs.
  • Demonstrated the potential of these LNPs for efficient intracellular protein delivery.
  • Highlighted the utility of microfluidics and design-of-experiment approaches in LNP formulation.