A platform method for simultaneous quantification of lipid and nucleic acid components in lipid nanoparticles

Area of Science:

• Pharmaceutical Sciences
• Analytical Chemistry
• Biotechnology

Background:

• Lipid nanoparticles (LNPs) are crucial for nucleic acid medicine delivery.
• Current analytical methods for LNP components are often separate and formulation-specific.
• A unified analytical approach is needed for LNP formulation development and quality control.

Purpose of the Study:

• To develop a single, versatile analytical method for simultaneous quantification of LNP lipid and nucleic acid components.
• To establish a platform method applicable to diverse LNP formulations and therapeutic cargoes.
• To address challenges in LNP process and quality control.

Main Methods:

• Developed an ion-pair reversed-phase HPLC method with UV and charged aerosol detection (CAD).
• Utilized a simple buffer with Triton and heparin for single-step simultaneous extraction of lipids and nucleic acids.
• Validated the method for quantifying 12 lipid species and three nucleic acid types (ASO, sgRNA, mRNA).

Main Results:

• Successfully separated and quantified both lipid and nucleic acid components in a single HPLC run.
• Achieved quantification recoveries between 90-110% for LNP components.
• Demonstrated the method's utility in assessing LNP purification recovery rates and quantifying co-loaded nucleic acids.

Conclusions:

• The presented HPLC platform method enables simultaneous analysis of diverse LNP components.
• This robust method simplifies LNP quality assessment and supports the development of nucleic acid therapies.
• The platform offers a valuable tool for process optimization and quality control in LNP manufacturing.