5 Key Questions On How To Accelerate mRNA Development
By Life Science Connect Editorial Staff
As the mRNA therapeutic industry continues to grow, stakeholders are working to solve key manufacturing challenges, including how raw materials and processes influence product quality and how molecular structure relates to function while also addressing the complexities of scaling production. Though other modalities’ existing analytical methods and manufacturing technologies may fall short for mRNA, investment in innovative techniques will help pharma meet the increasing demand for personalized therapeutics.
In a recent Advancing RNA Live Event, Accelerating mRNA-LNP Development, Dr. Tyler Goodwin, Head of Delivery Technologies and Technical Development at Tune Therapeutics; Dr. Adithya Nair, Research Associate at the University of Sheffield; and Dr. April Sena, VP of Technical Operations at Life Edit Therapeutics, discussed strategies for accelerating mRNA-LNP development via innovative analytical technologies, automation, and industry collaboration.
How Can Plasmid Production Keep Pace With mRNA Demand?
Due to their established use in AAVs and gene therapies, plasmids are one of the more mature components of mRNA development. However, as a critical raw material, plasmids remain technologically challenging due to their reliance on strategic cell selection and cell feeding techniques to develop an effective GMP master cell bank. While this flexibility is beneficial, it can be challenging for early-stage biotechs that lack the in-house expertise needed to define process requirements.
Despite these obstacles, the technology to accommodate plasmids is promising. “Instead of scaling bioreactors for better yields, the industry is focused on making healthier cells to improve plasmid quality,” said Goodwin, before adding that, “In-process analytics provide real-time monitoring of glycerol, glucose, lactate, and pH to control the plasmid fermentation process.” From a downstream perspective, developers must determine the necessary chromatography and purification steps to secure quality and maintain strong yields. Chromatography options continue to evolve to meet these complex needs.
Plasmid production is one of the most complex unit operations in mRNA-LNP development, and the industry continues to interrogate what quality of plasmid is necessary for product success. Given these challenges, Sena questioned whether e-coli-based plasmids would remain necessary: “I monitor amplification technology, and while it isn’t tailored to mRNA production, I think it will get there at some point, offering the potential to eliminate the cell-based plasmid component.”
How Are New Tools And Approaches Advancing IVT Performance?
Despite its central role in mRNA production, in vitro transcription (IVT) remains difficult to monitor and control in real time. Nair explained the challenge: “The first step to understanding kinetics is analyzing IVT at different stages to determine how much product has been synthesized. When using UV-absorbance techniques to determine yield, it is difficult to gain clear insight if both product and substrate give signals.”
Emerging IVT techniques aim to overcome this limitation by enabling separation of product and substrate for post-purification yield assessment, while newer methods explore bypassing purification altogether. Techniques using Raman spectroscopy and infrared spectroscopy (IR) can quantify product formation and substrate depletion in real time, providing clearer insight into reaction kinetics and enabling more precise process control.
Other IVT innovations include automated, plug-and-play bead kits for mRNA purification as well as modular approaches to mRNA synthesis that are especially applicable for personalized cancer immunotherapies. Modular setups are closed to control cross-contamination, reduce overhead, and increase the number of products produced in the same facility. The challenge is determining whether these modular setups are scalable outside of the personalized immunotherapy realm.
How Does Process Analytical Technology Need To Evolve?
To build an effective mRNA manufacturing process, developers must understand the process at different stages using effective process analytical technology (PAT). Because much of the PAT used in mRNA production is borrowed from more mature modalities, it was not designed to accommodate large-scale synthesis of nucleic acids in enzymatic processes. Legacy PAT can be adapted to determine certain variables, and different PATs can be combined with soft sensors to evaluate how a system is functioning. Ideally, the next wave of PAT will provide real-time feedback for all analytes.
Sena noted that, “At Life Edit Therapeutics, we bioengineer enzymes and use mRNA to express and screen them. In the future, PAT will hopefully get us to a point where we can be aware of how certain types of sequences affect key production parameters. We need to build a powerful data set and use AI to get there.”
How Is Digitization Transforming Process Control and Risk Management?
The industry is seeing a significant shift toward digitalization; for example, extracting information from real processes to make virtual models, such as digital twins, that can be used to control quality. Per Nair, “One step further would be advanced control mechanisms that flag issues before they reach the danger zone, allowing developers to take mitigation steps.”
Digitalization tools also pave the way toward greater automation, creating feedback loops to reduce operator errors, build greater risk reduction into workflows, and activate early-stage automation to support predictions for late stages. Automation in mRNA screening assesses the purity, process, and potential of different iterations. To leverage the power of this technology, developers must evaluate equipment and module vendors to determine if they offer automation potential.
Can COGS Be Meaningfully Reduced?
The answer is complicated, but yes, potentially. Capping reagents — required in high concentrations and not completely utilized — are the most expensive cost factor in the IVT reaction. Reusing or recycling capping reagents is one option to reduce COGS. Another is leveraging enzymes that do not require such high concentrations of a capping reagent. The issue, however, is that enzymes are proprietary, and as a result, developers are trading one proprietary material for another. Nair suggested the following: “Think about it from an engineering approach. How do I optimize the process with what is available rather than being dependent on something proprietary?”
To improve COGS, developers can utilize novel enzymes that drive upstream efficiencies, optimize yields, and reduce impurities to enable a more efficient and cost-effective downstream. Partnering with an mRNA CDMO will also help developers reduce COGS, streamline their process, and prepare a standardized regulatory package.
The Key To Continued Success
As mRNA developers gain a deeper understanding of how to adapt and implement new manufacturing approaches, stakeholder collaboration is key. Advancing the next generation of personalized therapeutics will depend on greater transparency and knowledge sharing, enabling the industry to collectively reduce risk, accelerate innovation, and build more efficient, scalable workflows.