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Related Experiment Video

Updated: Jun 11, 2025

Determining Four Components in a Lipid Nanoparticle RNA Delivery System by Liquid Chromatography Combined with Evaporative Light Scattering Detector
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RP-CAD for Lipid Quantification: Systematic Method Development and Intensified LNP Process Characterization.

Nicole Beckert1, Annabelle Dietrich1, Jürgen Hubbuch1

  • 1Institute of Process Engineering in Life Sciences-Section IV: Biomolecular Separation Engineering, Karlsruhe Institute of Technology (KIT), 76131 Karlsruhe, Germany.

Pharmaceuticals (Basel, Switzerland)
|September 28, 2024
PubMed
Summary

A new reversed-phase (RP)-charged aerosol detection (CAD) method offers comprehensive lipid nanoparticle (LNP) characterization. This technique enhances LNP quality control and process monitoring, identifying lipid deviations and losses during manufacturing.

Keywords:
bioprocessingcharged aerosol detectionintensificationlipid nanoparticlesmethod validationpower function valuereversed-phase chromatography

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Area of Science:

  • Analytical Chemistry
  • Biopharmaceutical Manufacturing
  • Nanotechnology

Background:

  • Lipid nanoparticles (LNPs) are crucial for nucleic acid delivery, but their quality assessment often focuses on particle size and encapsulation efficiency.
  • A need exists for comprehensive methods to characterize LNP composition and monitor manufacturing processes.
  • Current LNP characterization methods may not fully capture lipid content and process-related variations.

Purpose of the Study:

  • To develop and validate a holistic reversed-phase (RP)-charged aerosol detection (CAD) method for LNP and process characterization.
  • To apply the RP-CAD method to evaluate process parameters, specifically total flow rate (TFR), during microfluidic mixing.
  • To identify sources of lipid deviation and loss during LNP manufacturing.

Main Methods:

  • Development of an RP-CAD method with optimized power function value (PFV) using exploratory calibration.
  • Method validation including linearity (R² > 0.996), precision, accuracy, and robustness testing.
  • Application of the RP-CAD method to analyze LNP processing under varying TFR during microfluidic mixing.

Main Results:

  • The RP-CAD method demonstrated excellent linearity, precision, accuracy, and robustness for quantifying six common LNP lipids.
  • Analysis revealed a constant lipid molar ratio during microfluidic mixing, independent of TFR.
  • Deviations in lipid content were traced to lipid stock solution preparation, and lipid loss was attributed to post-mixing dialysis, both TFR-independent.

Conclusions:

  • The developed RP-CAD method provides a robust tool for lipid quantification throughout LNP processing.
  • This method enables detailed LNP characterization and process performance evaluation.
  • The findings highlight the potential for RP-CAD in optimizing LNP manufacturing and quality control, applicable to various LNP formulations and processes.