ArticleAnalytical chemistry2023
Separation of Plasmid DNA Topological Forms, Messenger RNA, and Lipid Nanoparticle Aggregates Using an Ultrawide Pore Size Exclusion Chromatography Column.
Article in Analytical chemistry, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Advancing Charge Detection Mass Spectrometry for Characterizing Large, Intact mRNA Constructs.Analytical chemistry · 2026Article
- Challenges and advances in analytical separation techniques for RNA-lipid nanoparticle therapeutics.Analytical and bioanalytical chemistry · 2026Review
- The Use of Enhanced Analytical Pipelines for the Characterization of Poly(A) and Poly(A)-LNP Formulation Critical Quality Attributes.Molecular pharmaceutics · 2025Article
- The impact of chemical reactivity on the quality and stability of RNA-LNP pharmaceuticals.Nature reviews. Chemistry · 2025Review
- Recent advances in analytical separation techniques for therapeutic oligonucleotides.Bioanalysis · 2025Review
- Characterization of intact mRNA-based therapeutics by charge detection mass spectrometry and mass photometry.Molecular therapy. Methods & clinical development · 2025Article
- Current Analytical Strategies for mRNA-Based Therapeutics.Molecules (Basel, Switzerland) · 2025Review
- Characterisation and analysis of mRNA critical quality attributes using liquid chromatography based methods.Journal of chromatography. A · 2025Review
- High-Throughput Quantification and Characterization of Dual Payload mRNA/LNP Cargo via Deformulating Size Exclusion and Ion Pairing Reversed Phase Assays.Analytical chemistry · 2025Article
- Keeping up with a Quickly Diversifying Pharmaceutical Landscape.ACS measurement science au · 2024Review
- mRNA vaccines in tumor targeted therapy: mechanism, clinical application, and development trends.Biomarker research · 2024Review
- Optimizing Messenger RNA Analysis Using Ultra-Wide Pore Size Exclusion Chromatography Columns.International journal of molecular sciences · 2024Article
- Not All Arms of IgM Are Equal: Following Hinge-Directed Cleavage by Online Native SEC-Orbitrap-Based CDMS.Journal of the American Society for Mass Spectrometry · 2024Article
- Comprehensive Impurity Profiling of mRNA: Evaluating Current Technologies and Advanced Analytical Techniques.Analytical chemistry · 2024Article
- Drug Product Development of Lipid Nanoparticle-Based mRNA Therapeutics and Vaccines.Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnologyReview
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Authors and funding
11 authors.
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No grant is acknowledged in the PubMed record.
Abstract
Health authorities have highlighted the need to determine oligonucleotide aggregates. However, existing technologies have limitations that have prevented the reliable analysis of size variants for large nucleic acids and lipid nanoparticles (LNPs). In this work, nucleic acid and LNP aggregation was examined using prototype, low adsorption ultrawide pore size exclusion chromatography (SEC) columns. A preliminary study was conducted to determine the column's physicochemical properties. A large difference in aggregate content (17.8 vs 59.7 %) was found for a model messenger RNA (mRNA) produced by different manufacturers. We further investigated the nature of the aggregates via a heat treatment. Interestingly, thermal stress irreversibly decreased the amount of aggregates from 59.7 to 4.1% and increased the main peak area 3.3-fold. To the best of our knowledge, for the first time, plasmid DNA topological forms and multimers were separated by analytical SEC. The degradation trends were compared to the data obtained with an anion exchange chromatography method. Finally, unconjugated and fragment antigen-binding (Fab)-guided LNPs were analyzed and their elution times were plotted against their sizes as measured by DLS. Multi-angle light scattering (MALS) was coupled to SEC in order to gain further insights on large species eluting before the LNPs, which were later identified as self-associating LNPs. This study demonstrated the utility of ultrawide pore SEC columns in characterizing the size variants of large nucleic acid therapeutics and LNPs.
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