The typical mRNA production process is a complex sequence of four key steps, each crucial for ensuring the quality and efficacy of the final product. The process begins with the expression of plasmid DNA (pDNA) in E. coli, followed by linearization and purification of the pDNA. This is succeeded by the in-vitro transcription (IVT) reaction, where mRNA is synthesized. The third step involves the purification of mRNA, ensuring its purity and quality. Finally, the nucleic acids are encapsulated into lipid nanoparticles (LNPs), which are essential for the in vivo delivery of mRNA vaccines and other nucleic acid-based therapies. Throughout these steps, various analytical methods need to be employed to monitor yields, recoveries and critical quality attributes. A multi step manufacturing process of complex biopharmaceuticals like these requires multiple orthogonal methods for a comprehensive understanding and control of the manufacturing. High purity of raw materials and meticulous in-process control during the IVT phase are critical for achieving high transcription yields, purer mRNA, and reduced consumption of costly reagents. The PATfix analytical platform for pDNA and mRNA enables such accurate and fast orthogonal chromatographic analytics. The encapsulation and subsequent downstream processing of RNA-LNPs is the last step in the manufacturing process, that is formulating the final drug product. LNPs are the most advanced non-viral delivery vehicles in nucleic acid delivery technologies, significantly advancing the possibilities of nucleic acid delivery. However, several challenges persist in their manufacturing and characterization. These include ensuring the correct formation and integrity of the drug product, achieving adequate recovery during purification while maintaining functionality, and producing a thoroughly purified and characterized final product. To address these challenges, purification using CIM monolithic columns has been developed, enabling fine-tuned separation and the production of a highly uniform and functional product.
