From global vaccines to personalized cancer therapies, mRNA is being considered across several clinical applications, which ultimately creates varying requirements in terms of production scale and dosage of the final mRNA product. This dictates the need for flexibility in the manufacturing process, as drug developers must determine how to consistently deliver high yields of quality mRNA in a timely fashion, while also balancing logistical and financial goals.
Working with an adaptable and collaborative CDMO partner can be critical to navigating industry challenges and finding the best solution to bring your mRNA vaccine or therapeutic to market. Thermo Fisher Scientific has responded to customer needs by offering a flexible solutions model for mRNA-based products (pictured above) that spans the entire operational value chain, from translational research to cold chain logistics.
Our end-to-end solutions align with each step in the mRNA workflow, with different options available to support varying production strategies and timelines. A plasmid DNA (pDNA) template is used as the starting material for mRNA synthesis via In vitro transcription (IVT), after which the resulting purified mRNA is encapsulated into a suitable carrier (e.g., lipid nanoparticles or LNPs) for efficient delivery and stability. While fermentation has traditionally been used to produce the pDNA templates, it involves lengthy production cycles and potential contamination risks. Synthetic DNA is currently being explored as a possible alternative to plasmid to alleviate some of these issues, potentially offering time and cost savings. Thermo Fisher Scientific can support either approach (Figure 1) and provide guidance on which option may be best suited for your product. We have established an in-house process for cell-free enzymatic DNA synthesis (EDS), or for customers preferring the fermentation approach, we partner with qualified vendors to provide a “white glove service” by handling all the plasmid work on the client’s behalf as a critical raw material. This includes procurement, quality management, technical support, etc.
After aligning on a strategy for DNA production, a linearized DNA template is generated as the starting material for the actual mRNA synthesis workflow (Figure 2). As a CDMO partner, Thermo Fisher Scientific can support mRNA manufacturing from process development work (milligram scale) through commercialization (kilogram scale). The early IVT process optimization is carried out using a systematic design of experiment (DOE) approach wherein we evaluate how different inputs/variables impact the process, ultimately helping to identify optimal conditions and minimize undesirable outcomes. In mRNA synthesis, critical goals include maximizing mRNA titer yields and reducing dsRNA content. mRNA modifications such as addition of a 5’ cap, modified nucleotides, and poly-A tailing are usually performed co-transcriptionally or post-transcriptionally. The synthesized mRNA transcripts are then purified using tangential flow filtration (TFF), precipitation and/or liquid chromatography (LC). Choice of purification method can be influenced by factors such as manufacturing scale, sample type, target purity, etc. The mRNA drug substance then undergoes in-process and release testing via a comprehensive suite of in-house analytical characterization services to assess identity, purity, quantity, potency, and safety. To continuously expand our innovative solutions, we have added a state-of-the art mass spectrometer (MS) to support mRNA capping efficiency method development, as well as other LC-MS techniques for quality control (QC).
Following mRNA synthesis, encapsulation (Figure 3) is a key step in the process to deliver mRNA into the target cells. Lipid nanoparticles (LNPs) are currently the most commonly used and well-studied nanocarrier for mRNA delivery; however, access to ionizable lipids, the key component of an LNP, is subject to a complicated IP landscape. When partnering with Thermo Fisher Scientific, we provide customers with access to proprietary lipids or we can optimize customers’ own LNP formulation leveraging DOE principles. Specific aspects of the LNP process optimization may include composition, mixing parameters, purification, cryopreservation, and sterile filtration. We also offer flexibility in equipment selection to provide multiple options for in-house manufacturing and analytics, and to accommodate tech transfer of existing processes.
To complete the process, the bulk LNP encapsulating the mRNA must be sterile-filtered and filled in the final container (e.g., vial, syringe, etc.), labeled, and stored at frozen temperatures (typically -20°C or -80°C) until it is ready for shipment. Thermo Fisher Scientific is uniquely positioned to support the end-to-end mRNA workflow with our co-location of mRNA manufacturing capabilities and sterile filling at our center of excellence in Monza, Italy, helping to minimize timelines and risks associated with handoffs to multiple vendors. Our model allows customers to leverage the full suite of integrated services or choose the specific ones that can help fill immediate gaps in their capabilities or capacity. In a complex and evolving market, we are a flexible and collaborative partner committed to ongoing innovations to help our customers accelerate development of new mRNA vaccines and therapeutics for patients in need. To learn more about our CDMO services for mRNA, please visit patheon.com/mrnaservices.