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Preparation, Administration, and Assessment of In Vivo Tissue-Specific Cellular Uptake of Fluorescent Dye-Labeled Liposomes
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In-Depth Characterization of PEGylated Liposomes: Using AF4 and HPLC-CAD in Tandem as a Strategy for Composition

Silvia L Appleton1, Guillaume Bucher1, Jessica Ponti1

  • 1European Commission, Joint Research Centre (JRC), Ispra 21027, Italy.

Analytical Chemistry
|June 25, 2026
PubMed
Summary

This study introduces a workflow for quality control of polyethylene glycol (PEG) liposome coatings, crucial for nanomedicine safety and efficacy. The method integrates microfluidics and advanced analysis for reliable PEG coating determination.

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Preparation and Characterization of Nanoliposomes for the Entrapment of Bioactive Hydrophilic Globular Proteins

Published on: August 31, 2019

Area of Science:

  • Nanomedicine
  • Pharmaceutical Sciences
  • Analytical Chemistry

Background:

  • Nanoparticle surface properties are critical for biological interactions and drug delivery.
  • Regulatory agencies require stringent quality control for nanoparticle surfaces, including polyethylene glycol (PEG) coatings.
  • PEG coatings are vital for nanomedicine efficacy and safety, notably in recent SARS-CoV-2 vaccines.

Purpose of the Study:

  • To propose and validate a workflow for overcoming challenges in liposome PEG coating.
  • To enable rigorous quality control of PEGylated liposomes for pharmaceutical applications.
  • To provide a reliable method for PEG coating determination and optimization.

Main Methods:

  • Integration of microfluidic synthesis with postinsertion of PEG.
  • Utilizing mild incubation conditions for PEG postinsertion.
  • Employing multidetector Asymmetric-Flow Field-Flow Fractionation (AF4-MD) coupled with High-Performance Liquid Chromatography-Charged Aerosol Detector (HPLC-CAD) for analysis.

Main Results:

  • Successful simultaneous analysis of size distribution and size-dependent parameters of PEGylated liposomes.
  • Efficient separation of PEGylated liposomes from unincorporated PEG-lipid micelles using AF4.
  • Demonstrated optimization of PEG postinsertion reaction conditions and validation of purification methods.

Conclusions:

  • The developed workflow offers a comprehensive and reliable method for in-depth liposome characterization, focusing on critical quality attributes.
  • The straightforward PEG coating determination is essential for postinsertion tuning and quality control.
  • The workflow is adaptable to other stealth coatings, serving as a valuable reference for nanomedicine development and regulatory compliance.