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Formulating and Characterizing Lipid Nanoparticles for Gene Delivery using a Microfluidic Mixing Platform
Published on: February 25, 2021
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Formulating and Characterizing Lipid Nanoparticles for Gene Delivery using a Microfluidic Mixing Platform.
Christina M Bailey-Hytholt1, Paroma Ghosh2, Julia Dugas2
1Biologics Drug Product Development & Manufacturing, Sanofi, Framingham, Massachusetts; Christina.Bailey-Hytholt@sanofi.com.
Journal of Visualized Experiments : Jove
|March 15, 2021
Summary
This study details a microfluidic method for creating lipid nanoparticles (LNPs) to encapsulate nucleic acids like mRNA and plasmid DNA. The reproducible technique precisely controls LNP formulation for effective drug delivery.
Area of Science:
- Biotechnology
- Nanomedicine
- Drug Delivery
Background:
- Lipid-based nanoparticles (LNPs) are effective drug carriers due to their size, biocompatibility, and high encapsulation efficiency.
- LNPs protect nucleic acids (RNA/DNA) from degradation and enhance cellular uptake.
- Key LNP components include ionizable lipids, helper lipids, cholesterol, and PEG-conjugated lipids.
Purpose of the Study:
- To develop and characterize a microfluidic platform for reproducible LNP formation encapsulating messenger RNA (mRNA) or plasmid DNA (pDNA).
- To investigate the influence of formulation and process parameters on LNP characteristics.
Main Methods:
- LNPs were formed by rapidly mixing lipids in an organic phase with nucleic acids in an aqueous phase using a microfluidic platform.
- Hydrodynamic size and polydispersity were measured by dynamic light scattering (DLS).
- Zeta potential determined surface charge, and encapsulation efficiency was quantified using a fluorescent dye assay.
Main Results:
- The microfluidic method demonstrated reproducible LNP self-assembly under laminar flow conditions.
- Characterization confirmed the ability to control LNP size, charge, and encapsulation efficiency.
- Representative results highlighted the impact of varying formulation and process parameters.
Conclusions:
- Microfluidic mixing offers a precise and reproducible approach for fabricating nucleic acid-loaded LNPs.
- This method facilitates the optimization of LNP formulations for advanced drug delivery applications.
- The study provides a foundation for developing tailored lipid nanoparticle-based therapeutics.
