Scaling up: moving from research to large-scale RNA production for mRNA therapeutics

As medical research continues to evolve, mRNA therapeutics have become a promising modality of many advancements in disease treatments. In fact, mRNA’s unique ability to prompt the cellular production of certain proteins has allowed the global scientific community to develop new mRNA-based approaches as a safe and cost-effective option for a breadth of applications.

Unsurprisingly, several mRNA therapies have been recently approved by the FDA—including COVID-19 vaccines—for large-scale production. To move into this manufacturing stage, mRNA therapeutics must first go through an extensive research and development process. By following the right steps and industry best practices, any lab can take their mRNA research from the bench to large-scale manufacturing.

Research and development: mRNA synthesis and purification

To begin mRNA synthesis via in vitro transcription (IVT), a DNA template with the given gene of interest (GOI) sequence is designed; this template serves as the blueprint for mRNA synthesis during IVT. IVT refers to the process in which an RNA polymerase enzyme transcribes the DNA template and synthesizes mRNA. After transcription, the template DNA can then be removed to ensure no interference with downstream applications.

Once completed, the mRNA is ready for the purification process. Clean-up kits are used to remove any contaminants, ensure high-quality mRNA, and ultimately minimize the risk of downstream interference. Post-transcriptional modifications (PTMs) can then be made, including the addition of a 5’ cap and polyadenylation (poly(A) tailing). A 5’ cap is key for protecting the mRNA against degradation and enhancing translation efficiency; a poly(A) tail helps stabilize the mRNA and aids in its translation as well. The workflow should be carefully designed for future scalability.

Process development

Because the research and development stage operate with laboratory-scale experiments, the move to industrial-scale manufacturing can be challenging. An effective transition first requires small batch production to first ensure the mRNA can be replicated on a larger scale without affecting its quality.

In addition, the process will also need high-quality (HQ) materials with consistent purity and functionality that has been confirmed by rigorous testing. Many suppliers offer ‘transitional’ high quality (HQ) enzymes with features comparable to GMP grade quality, facilitating a smooth transition to GMP-grade manufacturing. During the transition from research to mid-scale mRNA production, the safety and efficacy of the IVT mRNAs should be consistent throughout the scaleup process. The use of large package size of enzymes and other high-quality materials helps maintain consistency and minimizes lot-to-lot variations.

Large-scale manufacturing

Transitioning from process development to large scale manufacturing is not a simple or linear process. It requires careful evaluation that the production process is efficient, consistent, and that the product meets the requirements and expectations. Utilizing a contract development and manufacturing organization (CDMO) allows for access to specialized expertise and facilities, ensuring high quality production is maintained. CDMOs also offer flexibility, enabling researchers to quickly adapt to market demands and streamline the transition.

Getting started

Transitioning mRNA therapeutic research from a small research scale to the large-scale production can be a long process with many steps. Thankfully, there are a variety of tools that can help simplify each stage for any interested party, no matter their level of production scale.

Learn more about products and services for the research, development, and manufacturing stages today.