Optimization, Scaling and Characterization
Further optimization and characterization of LNPs are needed to select the desired formulation that yields optimal encapsulation efficiency, stability, cellular targeting and uptake, and potency. This information is crucial for scaling up LNPs that facilitate effective cellular uptake and promote the release of mRNA in target cells, leading to robust protein expression.
With the utilization of the Beckman Coulter Life Sciences Optima XPN and density gradient purification, LNP samples can be optimized for functionality and evaluated for heterogeneity, providing valuable insights into the composition and characteristics. Additionally, for characterization, Beckman Coulter Life Sciences has several products, including the Optima AUC Analytical Ultracentrifuge, which allows for precise quantification of LNP loading states, hydrodynamic radius and RNA copy number distribution.
Phenomenex Biozen Columns and LC Accessories enable analytical quality by design to ensure high-quality data during optimization and characterization. The SCIEX ZenoTOF 7600+ system provides deep insights into the structure of lipids, including lipid impurities, with excellent quantitative performance for data-driven decision-making. The SCIEX BioPhase 8800 system and PA 800 Plus system utilize capillary electrophoresis to inform about mRNA integrity, purity, size and LNP encapsulation efficiency.
Effective data management allows for the systematic recording of experimental conditions, results and observations. This aids in identifying optimal conditions for LNP formation, scaling up the process, and characterizing the particles in terms of size, charge and other relevant parameters. Automated data analysis and seamless integration of results across multiple assays are crucial for successful scale up efforts. Genedata and IDBS provide scalable, enterprise-level cloud software products from R&D through manufacturing for assured confidence in both GxP and non-GxP environments.
Genedata Biopharma Platform automates routine and custom LC/MS and LC/UV/MS data analysis for deeper characterization to accelerate large-scale high-throughput analyses. It also automates and centralizes the analyses of plate-based assays like dynamic light scattering (DLS), TNS ionization, mRNA encapsulation, toxicity, cell trafficking and automated ELISAs. In addition, it integrates data from optimization to characterization, supports analytical development, and tracks raw materials, samples, batches and all assay results at the enterprise level. Out-of-the-box, the Genedata Biopharma Platform can support optimization, scaling and characterization of LNPs.
Capabilities within IDBS Polar software allow scientists to design, execute and analyze experiments within a single platform to understand formulation composition, material genealogy and determine which process parameters are most influential based on statistical analysis. This allows scientists to improve reproducibility to reduce the number of experiments that are needed to gain process understanding and enables actionable insights to optimize, scale and characterize LNP development processes.
Based on the characterization and efficacy assessment results, iteratively optimize the lipid composition and ratios, refining the formulation until the desired performance metrics are achieved. If the final formulations are intended for clinical use, ensure they comply with regulatory guidelines for safety and efficacy.