A FRET-based toolkit for quantifying lipid incorporation into nanoparticles

Area of Science:

• Biotechnology
• Materials Science
• Nanotechnology

Background:

• Lipid-based nanoparticles are promising for drug delivery due to their modularity and chemical diversity.
• Current tools for assessing lipid assembly and distribution within nanoparticles are limited, hindering rational design.
• Understanding lipid integration is crucial for developing functional lipid-based drug-delivery systems.

Purpose of the Study:

• To develop and validate a Förster resonance energy transfer (FRET)-based approach for quantifying lipid incorporation in liposomes and lipid nanoparticles (LNPs).
• To create a theoretical model for predicting lipid distribution within nanoparticles and experimentally validate it.
• To enable quantitative analysis of individual lipid components in complex nanoparticle formulations.

Main Methods:

• Development of a modular and adaptable Förster resonance energy transfer (FRET)-based assay for lipid quantification.
• Creation and experimental validation of a theoretical model for labeled lipid distribution in nanoparticles.
• Application of the FRET assay to monitor lipid post-insertion and quantify phospholipid incorporation, including plasmalogens.
• Extension of the FRET toolkit to study lipid incorporation in lipid nanoparticles (LNPs) and confirmation with Fluorescence Nanoparticle Tracking Analysis (F-NTA).

Main Results:

• The FRET-based approach successfully quantifies lipid incorporation in both liposomes and LNPs.
• The theoretical model accurately describes labeled lipid distribution, guiding experimental design.
• The study quantified the incorporation of various phospholipids, including challenging plasmalogens.
• Lipids were found to incorporate uniformly into LNPs without disruption, confirmed by F-NTA.

Conclusions:

• The developed FRET-based toolkit provides a quantitative method for analyzing lipid components in nanoparticles.
• This approach addresses a critical gap in nanoparticle characterization, facilitating the design of advanced lipid-based therapeutics.
• The toolkit accelerates the rational design and development of next-generation drug delivery systems.