Single-use technology is transforming mRNA-LNP manufacturing

By uncovering crucial insights into genetic links to disease and drug response, genomic medicine is harnessing the power of DNA to revolutionize healthcare. A deeper understanding of genetic predispositions drives personalized medicine, where patient-specific treatment plans and targeted therapies that offer optimized outcomes with fewer adverse effects can be developed. As technology advances, genomic medicine is poised to unlock new possibilities for precision healthcare and transform how we approach disease management and prevention.

The successful use of lipid nanoparticle (LNP) technology has advanced genomic medicine and supported the development of mRNA-based formulations. With the COVID-19 vaccines as a notable example, LNPs have attracted much attention and success in the field of mRNA-based vaccine and drug development.

Structurally, LNPs typically consist of four components ― ionizable lipids, phospholipids, cholesterol, and PEGylated lipids ― that self-assemble into nanoparticles and develop a protective shell around the therapeutic cargo or “payload”, such as mRNA. This protective encapsulation carries the payload through the body to its target cells, making LNPs an effective drug delivery system for various applications, including cell and gene therapy. LNP technology has the potential to fundamentally alter how we prevent, identify, and cure diseases.

Single-use technologies accelerate genomic medicine development timelines

Speed to market is critical, but producing a commercial mRNA-LNP drug product often comes with difficulties that take time to overcome. Single-use technology (SUT) has gained attention in the field for its ability to facilitate a reliable, robust, and controlled LNP manufacturing process while helping to maintain sterility ― a key factor when manufacturing LNPs, as contamination can negatively impact their safety and efficacy.

Joe Makowiecki, Business and Product Development Leader for Enterprise Solutions at Cytiva, says the situation now reminds him of the early days of adoption of single-use technology for monoclonal antibody (mAb) manufacture.

"We had to do a lot of convincing on the benefits of single-use equipment over the existing stainless-steel (clean and re-use) infrastructure. There was a lot of cynicism and questions, like: 'Why would you want to use single-use? What do you do with the plastics that are generated? Does it have the same capabilities as the existing stainless-steel systems? And beyond that, what does it give you that differentiates it?' These were the conversations we were having back then. It was a stainless-steel world, and we were introducing a new paradigm."

Almost 20 years later, the benefits of SUT are well recognized in the manufacture of monoclonal antibodies and biopharmaceutical manufacturing in general. As RNA technology evolves from mRNA to self-amplifying RNA (saRNA) and circular RNA (circRNA) and the encapsulation technologies also become more targeted and complex, Makowiecki says operational flexibility in the manufacturing process will become a prerequisite for RNA manufacturers and that modularized single-use equipment will play a critical role.

Sterility is a significant focus when producing genomic medicines, as contamination can lead to loss of therapeutic activity, regulatory delays, or rejections due to safety concerns. For example, ribonucleases (RNases) from human and animal tissues and microbial sources during sample preparation, RNA extraction, purification, or formulation can lead to RNA degradation.

Single-use components, such as disposable mixers, tubing, and filters, minimize the risk of contamination. They also reduce the need for cleaning, maintaining, and validating reusable equipment, benefiting biopharmaceutical companies by decreasing the need for time-consuming and labor-intensive work. In addition to the reduced risk of contamination, enhanced product quality and safety and simplified validation processes help ensure compliance with regulatory requirements, accelerating speed to market.

Cost efficiency is another critical factor driving the adoption of SUT. Although single-use components may initially be more costly than reusable equipment, the overall cost-efficiency becomes evident over time. With disposable components, there's no need to invest in equipment cleaning, maintenance, and validation procedures. Additionally, the risk of batch failure and associated downtime is reduced, saving time and money in the long term.

Workflow efficiency is also crucial, given the pace at which genomic medicine is advancing. Multi-product and multi-process workflows allow the manufacture of multiple mRNA products in the same facility, and using single-use equipment for cGMP production supports efficient changeover between products. The platform approach of single-use cartridges, such as those found in NanoAssemblr™ formulation systems, accommodates various manufacturing setups for efficient facility utilization across products and scales.

The use of SUT in mRNA-LNP manufacturing means that extractables and leachables (E&L) are also a consideration, with manufacturers following guidelines such as the United States Pharmacopeia (USP) chapter 665 and the BioPhorum protocol for testing of product-contacting plastic components. Testing considerations for mRNA vaccines and therapeutics include modeling exposure based on dose, such as a low-dose regime for a prophylactic vaccine or a high-dose regime for a protein replacement therapy.

Single-use technology supports scale-up and scale-out manufacturing

As the demand for LNPs and mRNA-based therapeutics grows, it is crucial to have a scalable manufacturing process that can quickly adapt to different production volumes.

SUT offer consistent and reproducible performance, minimizing the potential for variability and the risk of batch failure when transitioning from preclinical to GMP manufacturing. Unlike stainless-steel equipment which requires a larger investment, SUT also allow manufacturers to easily scale production volumes up or down by adjusting the number of single-use bioreactors, mixing bags, or other single-use components. This equipment modularity was a significant factor in swiftly and efficiently setting up manufacturing lines across the globe during the COVID-19 pandemic (1). Also, its advantage in expediting technology transfer ― whether new lines are added inside an existing facility or new facilities are established elsewhere ― makes SUT an efficient, reliable, and attractive option for manufacturers looking for mRNA-LNP production at various scales.

Cytiva’s NxGen™ technology empowers its NanoAssemblr™ family of products to reproducibly generate optimal particles through a single-use mixer across scales, from the preclinical NanoAssemblr Ignite™/Ignite+™ and NanoAssemblr Blaze™/Blaze+™ systems to clinical and commercial systems. The NanoAssemblr™ commercial formulation system is an automated, single-use system for the clinical and commercial production of LNPs under cGMP conditions. Its single-use flow path minimizes the need for sanitizing and performing cleaning validation, enabling efficient changeover between production runs while minimizing the risk of cross-contamination and supporting multi-product manufacturing in GMP facilities.

With the systematic development of new technologies, SUT provides a flexible and adaptable platform for integrating innovations quickly. By adopting single-use technologies and platforms into their workflows, biopharma manufacturers can reduce the time required for facility setup, cleaning, and validation, enabling faster turnaround times and accelerating the production of mRNA-based vaccines and therapeutics.