Lipid nanoparticles are uniquely designed to encapsulate a variety of therapeutical agents offering a safer and more adaptable alternative to traditional viral systems. While LNPs are transformative, their unique properties necessitate specialized downstream processing to ensure quality and effectiveness for in vivo applications. Post-formulation, LNPs require downstream processing to make the formulation applicable for in vivo applications and monitoring of CQAs.
CIM® OH monolithic columns are uniquely suitable for LNP purification due to the low shear stress imparted of the purely laminar flow and a wide selection of chemically modified surfaces.
Current methods for analyzing LNPs require complex sample handling due to their complicated composition and structure, making characterization challenging. PATfix® LNP Switcher Platform – a comprehensive analytical chromatography solution was developed to tackle these challenges effectively.
Beyond Filtration: Chromatographic Purification of LNP
Our innovative chromatographic purification process for LNP-based biopharmaceuticals delivers uniform and functional drug products, ensuring improved quality and safety. With high recoveries and full scalability, CIM® technology is designed to meet diverse purification needs. Ready-to-use columns accommodate everything from microgram quantities for screening to tens of grams of encapsulated nucleic acids.
The chromatographic method using CIM monolithic OH columns for RNA-LNP products achieves high recovery and product stability, effectively removing ethanol and unencapsulated RNA. Compared to standard processes, our approach delivers higher recoveries (>90%), more uniform particles, and enhanced protein production activity, confirmed by cell-based assays.
The Chromatographic Solution for Studying LNP Formulations
The complex composition and structure of LNPs pose significant challenges for their characterization and analysis. PATfix LNP Switcher Platform is an all-in-one analytical chromatography solution that addresses these challenges, enabling the investigation of multiple critical attributes for LNP drug products, without extensive sample preparation or dye requirements. Suitable for analytics during LNP process development and investigation of drug product stability post-formulation.
How Does the PATfix LNP Switcher Platform Work?
The LNP Switcher is equipped with two analytical columns of different modalities, independent pumps and multiple detector setup. This hardware configuration enables the monitoring of key quality attributes during the LNP production process.
Watch the video to learn how PATfix LNP Switcher Platform enables multiparameter LNP analytics and enables determination of critical LNP quality attributes such as
- encapsulation efficiency and yield,
- nucleic acid integrity,
- and average particle size.
- Load:
Direct injection of LNP formulations; no sample pre-treatment, only dilution with loading buffer.
- Analysis of Free Nucleic Acids:
HIC column with 6 μm channels selectively binds LNPs, separating free nucleic acids, which are analyzed for quantity, size, and integrity on the CIMac SDVB column.
- LNP Elution:
The elution of LNPs is detected by the MALS detector, where size and heterogeneity of particles is determined.
- Analysis of Encapsulated Nucleic Acids:
Reverse phase analytics allows LNP deformulation and payload release in one step, with UV-Vis detector analysis of nucleic acids for: concentration, type, size, integrity, and RNA-lipid adducts.
Control the Whole Production Process from pDNA to LNP
PATfix is a process development tool designed for fast, high-resolution analysis of biomolecules and complex in-process samples. The PATfix platforms are user-friendly and come with pre-developed and pre-validated methods.
PATfix pDNA Analytical Chromatographic Platform
- Lysis optimization
- Monitoring and optimization of the purification process
- pDNA isoform concentration
- Optimizing plasmid linearization kinetics
PATfix mRNA Analytical Chromatographic Platform
- Controlling IVT reaction
- Monitoring & optimization of the purification process
- Quantification of mRNA integrity and dsRNA detection
PATfix LNP Analytical Chromatographic Platform
- Monitoring encapsulation efficiency of different nucleic acids
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Determination of payload integrity
- Monitoring LNP size and distribution
The PATfix analytical platforms are designed to simplify analytical chromatography and make its application easier for process development scientists. The platforms offer:
- A pre-set system with recommended detectors
- Fully optimized and validated analytical methods (and guidelines for buffer and sample preparation, and SOPs)
- Analytical columns that best fit the analytical needs
- User-friendly PATfix Software
