In a November 2022 webinar, Linda Mathiasson (strategic customer lead at Cytiva) discussed the significant promise of messenger ribonucleic acid (mRNA), its role in fighting genetic and infectious diseases, and how its production can be scaled to meet the needs of different applications, ranging from large-scale vaccination to production of individual therapies.

The Presentation
Thanks to the rapid development of mRNA vaccines for COVID-19, the biopharmaceutical industry has gained momentum toward realizing the technology’s potential. The speed, flexibility, and low-cost scalability of mRNA platforms bring distinct benefits for developing vaccines against Zika virus, influenza, and diseases caused by emerging pathogens. The technology could play a role in developing cancer vaccines, cell therapies, and protein-replacement drugs. It could also be used as a delivery method for gene-editing products.

Manufacturing processes for mRNA span a broad range of scales. Large-scale operations such as those for vaccine manufacturing require high-throughput processes that provide strong yields at the requisite level of quality. Such parameters remain important at small scales, but considerations such as cost of goods (CoG) and patient tracking arise.

Mathiasson noted that if many small-scale processes run in parallel, each needs careful collection and management of process data. Integration, automation, and digitalization are key when running many smaller batches in parallel, such as in personalized therapies that may run and release thousands of batches in a year. Innovation in this area has the potential to influence larger-scale manufacturing in the future. Mathiasson highlighted different ways of handling process diversity at small scales, each with different drivers. Manufacturers can scale down separate unit operations and connect them physically or digitally. Alternatively, companies can apply a fully integrated approach.

Mathiasson calculated that a 2-L production line might output about 200 g of mRNA per year, assuming a titer of 5 g/L and a 46% yield. As process volumes decrease, so do run times, enabling the ability to produce many more batches per year. In this case, a ballroom-facility approach could be considered when using closed systems. If single-use flowpaths are implemented, turnaround times will be accelerated because there will be no need to clean between batches.

Companies can leverage flexible single-use equipment throughout the mRNA workflow, which covers a broad range of scales with different flow kits. That enables the manufacturing line to adapt to different processes.

Mathiasson presented data showing that an in vitro transcription (IVT) reaction can be scaled from 1 mL to 10 mL in test tubes to 150 mL in a single-use rocking bioreactor that can be scaled further to 25 L with consistent output. It is also of great importance to control ribonuclease (RNase) activity in consumables. Mathiasson presented a case study with data showing low consumable RNase activity, comparable to that of negative controls.

Cytiva recently launched the ÄKTA ready 450 single-use chromatography system for use in small-scale manufacturing of mRNA and other modalities such as viral vectors and monoclonal antibodies (MAbs). The system has low hold-up volume and slow flow rates to support small-scale purification, and it is designed as a closed system to reduce contamination risk. The system is flexible, scalable, and small enough to use on a laboratory benchtop.

Mathiasson noted that small-scale production magnifies the need for digitalization in tracking batch and patient data, eliminating reliance on four-eye verification and labor-intensive manual logging. Digitalization is also useful for compliance considering regulatory agencies’ drive toward electronic submissions.

Questions and Answers
You mentioned that innovations in mRNA manufacturing are likely to begin with small-scale processes. Can you elaborate on that? Intensification and integration of workflows will be key, as will maintaining the flexibility to adapt to different products, with personalized cancer vaccines serving as one example. Automation, digitalization, patient tracking, and integration will be more urgent for such products.

Will the need for single-use components continue to grow as manufacturing technologies move toward smaller batch sizes and higher batch numbers? Yes, that is likely. Single-use systems will be especially beneficial when they are used in parallel processes. You can close those processes and decrease their turnaround times.

Find the full webinar online at www.bioprocessintl.com/category/webinars.