Sara Sousa Rosa, PhD is a research fellow at UCL Biochemical Engineering, in the UCL-Cytiva Center of Excellence. Her current research focuses on the optimization of manufacturing processes of mRNA vaccines, from production through to formulation. Prior to her current role, Sara worked at iBET, where she developed downstream processing strategies for complex biopharmaceuticals, including viral vectors and virus-like particles (VLPs) for clinical applications. She joined IBB to contribute to developed purification strategies for monoclonal antibodies, plasmid DNA, and extracellular vesicles (EVs). She was awarded a PhD scholarship by the Portuguese Science Foundation for her work on optimizing mRNA vaccine manufacturing processes, carried out in collaboration with IBB (University of Lisbon) and UCL Biochemical Engineering.
About this event:
Date: June 3, 2025
Time: 11 am EDT | 8 am PDT | 5 pm CEST
Duration: 15 minutes
mRNA vaccines have emerged as a promising technology for a number of applications, from prophylactic and cancer treatments to metabolic and genetic diseases. This is mostly attributed to the high precision and perceived safety as well as flexible manufacture of these vaccines. mRNA is produced in a cell-free system, in vitro transcribed by a RNA polymerase, that uses nucleotides and DNA template as substrates. This cell-free reaction can be optimized achieving yields of over 14 gRNA.L-1 under one hour. With ever increasing mRNA titres and RNA modalities, the production of product- (e.g. double-stranded RNA, dsRNA) and process-related (e.g. DNA template, and free NTPs) impurities will increase.
In particular, the removal of dsRNA is critical due to the strong immune response in the presence of this impurity, as well as to the translation inhibition and uncontrolled immune-inflammatory responses. To ensure the removal of this impurity and a high pure mRNA, multiple unit operations are employed such as filtration and chromatography. However, a complete separation of mRNA from dsRNA is difficult to achieve owing to their physicochemical similarity. Thus, to enable mRNA technology to reach its full potential, establishing cost-effective and flexible manufacturing processes are required.
This webinar will showcase the use of multimodal chromatography for the separation of mRNA from challenging impurities such as dsRNA. By focusing on impurity profiles, binding conditions were optimized to allow flow-through of major process and product-related impurities, while maximizing mRNA binding. Further optimization binding conditions led to improved mRNA recoveries. Final process conditions allowed to obtain mRNA recovery yields of >80% and purity >88%. Additionally, over 65% of the dsRNA is removed, and no detectable amounts of DNA are found in the purified samples. This streamlined purification method enables the isolation of high quality mRNA in a single chromatography step, eliminating the need for multiple purification stages within the mRNA manufacturing platform. This approach has the potential to significantly reduce processing time and lower manufacturing costs, making the platform more attractive for a range of applications.
Key Takeaways:
mRNA vaccines are produced in a simple, flexible, and high-yield cell-free process, but deliver a high number of process and product-related impurities
Multimodal chromatography presents higher selectivity to separate impurities that are physiochemically similar to mRNA compared to affinity chromatography
Optimization of binding and elution conditions allows to deliver a one-step process where a final, highly pure mRNA is obtained.
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