mRNA Revolution:

Innovate case study

What is the role of Innovate UK in supporting the development of mRNA manufacturing in the UK?

As part of the Transforming Medicines Manufacturing Programme, which is the current Innovate UK programme for supporting the new medicines manufacturing sector in the UK, they have run three competitions to cover the broad remit of RNA therapeutic manufacturing innovation.

The origin of those competitions goes back to an agreement between Novartis and the Department of Health around inclisiran, which is an oligonucleotide therapy for high cholesterol. This therapy is an alternative to statins and offers the benefits of being an injection taken twice a year rather than a tablet taken daily. It has the potential to save significant cost in the NHS by reducing the number of deaths related to heart disease associated with high cholesterol.

Currently those types of therapies cannot be manufactured effectively at large scale, certainly not sustainably at large scale with the established process. With a number of company pipelines looking increasingly at RNA therapeutics and mRNA therapeutics in particular, Innovate UK saw a growing need and, through the Medicines Manufacturing Challenge Fund, supported a project to look at a next generation manufacturing process for oligonucleotides.

Through COVID-19, where mRNA vaccines came to the fore and the UK had a very successful programme in the development of the Oxford AstraZeneca vaccine there was a recognition that it would be valuable to invest in parallel in technologies enabling mRNA process development, therapeutics development, vaccines development and delivery technology development.

Innovate UK realised that for the UK to adopt this world-leading position, much could be learnt from the Cell and Gene Therapy Catapult (CGTC) in terms of the position that has provided Innovate UK in cell and gene therapy expertise. This would be a good opportunity to start to make investments and adopt a leading position in mRNA type technologies as well, for global benefit, but developing manufacturing capability and expertise in the UK.

What grants have already been awarded?

CPI’s Intracellular Drug Delivery Centre will help predict the stability, efficacy, performance and any potential adverse reactions of RNA vaccines and therapeutics. This will help unlock the potential of RNA-based medicines, creating greater access to cost-effective vaccines and therapeutics. This is a £10 million investment over three years.

Innovate UK have funded projects that are focused on a broad section of RNA therapeutics and vaccines including a number of mRNA technologies but also other RNA and oligonucleotide based technologies.

Moderna case study

What does the company do?

Moderna creates medicines that aim to deliver the greatest possible impact to people by using mRNA technology. The COVID-19 pandemic proved that mRNA was the fastest route to developing highly effective vaccines and since 2010, Moderna have been using this technology to develop and test a wide range of medicines that span a variety of therapeutic areas, such as infectious diseases, immuno-oncology and rare diseases. The speed, scale and flexibility of mRNA allows us to accelerate the discovery of new medicines, and get them to those who need them, faster.

Tell us about Government facilities in the UK and supplier agreement. 

The strategic partnership between Moderna and the UK Government, signed in December 2022, will boost the country’s research and development capabilities with the construction of the Moderna Innovation and Technology Centre (MITC) in Harwell, Oxfordshire. This state-of-the-art R&D and manufacturing centre will be at the heart of the partnership to develop and manufacture innovative vaccines for respiratory diseases. Production capacity at the centre can be scaled up in the event of a pandemic, offering the potential to develop up to 250 million vaccines per year, thus massively enhancing the UK’s ability to cope with and respond to future public health threats. 

Why set up in the UK?

Moderna is excited to be making a significant investment in the UK and is working together with a range of partners, to support the UK to unleash its full potential and cement itself as a life sciences superpower. Thanks to the UK’s thriving biomedical sector, Moderna believes that it is an excellent location to carry out research and development. Moderna are looking forward to working with leading scientists on the next generation of treatments and are committed to upstream collaboration, co-creating with partners, staying attentive to their needs, and maintaining a seamless partnership in clinical research. 

The work Moderna are doing in the UK would not be possible without the UK Government who recognise the importance and value of life sciences, the efforts of the NHS, world-leading scientific research institutions, a progressive regulator in the form of the MHRA (Medicines and Healthcare Products Regulatory Agency) and many others. Moderna have already built close relationships with Merit, for example, a construction specialist based in Cramlington, Northumberland who are partnering with Moderna to develop a biomarker lab that will form part of the MITC. They are onshoring their supply chain in the UK as much as possible and look forward to more partnerships in the coming years.

What are the challenges? 

The strategic partnership will operate at the intersection of the public and private sector and its success will hinge on the seamless collaboration between numerous partners, each bringing unique expertise and perspectives.

The skills shortage in the life sciences sector poses a challenge, particularly given the specialised nature of mRNA technology. However, this also presents a unique opportunity for Moderna to invest in the development of the UK’s talent pool, fostering the next generation of a highly skilled workforce.

The scale and complexity of Moderna’s pipeline of clinical trials will also demand careful planning with system partners. Moderna are committed to working with the UK Government to implement the recommendations of Lord O’Shaughnessy’s report on commercial clinical trials and to create a more innovative, inclusive, and digitally enabled clinical trial ecosystem.

UK-MRNA-2300072 Date of preparation: August 2023

SiSaf case study

What does the company do?

SiSaf is an RNA delivery and therapeutics company. Its proprietary Bio-Courier^® technology addresses the limitations of other nucleic acid delivery technologies through the hybridisation of organic materials with inorganic bioabsorbable silicon. The company’s business model is to maximise the potential of its technology by licensing to industry partners and by developing an in-house pipeline of RNA therapeutics. SiSaf’s most advanced programmes are RNA treatments for rare genetic disorders, but the company has the strategic ambition to expand its focus to other therapeutic areas, including oncology.

How does the technology work? 

Lipid nanoparticles (LNP) have become the standard delivery system for mRNA. Bio-Courier^® silicon stabilised lipid nanoparticles (sshLNP) are the next generation of lipid nanoparticles combining organic material – lipids – with inorganic material – bioabsorbable silicon. The silicon matrix stabilises both the lipid components and the RNA payload, reducing the need for ionisable or cationic lipids, PEGylated lipids, and cholesterol. This improves safety, targeting, and transfection. Unlike conventional lipid nanoparticles, sshLNP can be manufactured empty and loaded with the RNA later, at the desired point and time of use.

How is this an improvement on existing delivery technologies?

The stabilisation of both lipid components and RNA payload through silicon, as well as the ability to manufacture the carrier without RNA, overcome the key limitations of conventional LNP delivery systems for nucleic acid.

In conventional LNPs, the nucleic acid payload must be introduced early in the production process during the initial formation of LNPs prior to subsequent purification and fill/finish steps. This restricts batch sizes in commercial manufacturing due to the chemical lability of RNA, it affects the quality of the product that ultimately reaches the patient, and requires an ultra-cold chain for storage and shipping. In Bio-Courier sshLNPs, the nucleic acid can be introduced after the manufacture of the sshLNP particles. As a result, RNA encapsulation and fill/finish operations can be separated from carrier manufacture by considerable time and distance, readily permitting on-demand preparation of customised RNA-loaded LNPs on any scale.

In addition, Bio-Courier technology offers significant advantages over conventional LNP technology regarding stability, safety, targeting, and transfection.

Stability
The use of the positively charged silicon matrix to bind and stabilise the RNA payload reduces the need for cationic/ionisable lipids, avoiding the risk of premature leakage of RNA from the nanoparticles as the positive charge from the lipids decays. The improved physical stability also translates into improved transfection efficiency.

Safety
Existing approved LNPs rely on PEGylated lipids to protect RNA against enzymatic degradation, however, PEGylated lipids can induce anti-PEG antibodies and provoke an immune response. Due to their silicon-stabilised design, Bio-Courier sshLNPs are less reliant on PEGylated lipids than conventional LNPs, to the extent that some formulations are entirely free of PEGylation.

Targeting
LNPs have a strong tendency to accumulate in the liver. Biodistribution studies with Bio-Courier sshLNPs have demonstrated minimal accumulation in the liver and Bio-Courier formulations can be customised to provide controlled release and targeting of specific cells and tissues.

What are the challenges?

Opportunities for RNA medicines have advanced hugely in recent years. That being said, RNA remains a new modality with all of the associated development, manufacturing and regulatory challenges. SiSaf believe delivery technology helps move the field forward positively.

The patent landscape in RNA therapeutics including delivery platforms is complicated. However, SiSaf are confident of the strength of our IP (Intellectual Property) portfolio and both our freedom to operate and our ability to license our technology to others. 

As a small but ambitious company, SiSaf needs to focus its resources on demonstrating the power of its technology and to attract external partnerships and investment to progress clinical programmes. We are proud that our lead programme SIS-101-ADO, an siRNA therapeutic for patients with Autosomal Dominant Osteopetrosis Type 2 (ADO2), has been granted both Orphan Drug Designation and Rare Pediatric Disease Designation by the U.S. FDA (Food and Drug Administration). There are currently no approved treatments for Osteopetrosis ADO2 and no other treatments currently in clinical trials. If approved, SiSaf’s SIS-101-ADO would thus be the first treatment for Osteopetrosis ADO2 and could provide life-altering benefits for those who suffer from this debilitating disease. Their fast follower programme, SIS-102-ACH, would offer the first treatment of Achondroplasia that targets the genetic root cause of the condition. SiSaf also have partnered programmes to treat Type II Corneal Dystrophy and an undisclosed programme using Bio-Courier for topical delivery.

SiSaf have raised significant funding to date. They are committed to take their lead programmes through early clinical development, and thus anticipate a series C in due course.

Dr Suzanne Saffie-Siebert

Founder & CEO, SiSaf Ltd
Inventor of Bio-Courier^® technology

RNA-based medicines are an incredibly exciting and promising new field of drug development. But the inherent instability of RNA means that delivery systems are key to the success or failure of RNA drugs.

At SiSaf, we have been developing and optimising Bio-Courier technology for 15 years. We have generated a large body of in vitro and in vivo data that show the safety and efficacy of our technology and we have created a strong IP portfolio that gives us extensive freedom to operate. I believe our innovative technology can play a key role in translating RNA-based medicines into clinical reality on a global scale.”

CPI case study

What does the company do?

CPI is a technology innovation catalyst. They catalyse the adoption of advanced technologies and manufacturing solutions that benefit people, places and planet. CPI is a social enterprise, partnering with industry, academia, government and the investment community to deliver healthcare and sustainability innovations. 

Within pharmaceuticals, CPI offer world-leading facilities and expertise in small molecules, biologics, oligonucleotides, and complex medicines to accelerate drug development and scale-up. They host the largest pool of RNA biomanufacturing experts in the UK and offer complementary drug delivery systems like lipid nanoparticles.

CPI supported the UK Vaccines Task Force’s COVID-19 response with their world-leading expertise in the development of mRNA vaccines, working with partners such as Imperial College London to support the drive to develop, scale up, manufacture and supply vaccine candidates.

As part of this, CPI developed and built a new facility, The RNA Centre of Excellence, next to their National Biologics Manufacturing Centre in Darlington. This was designed to be a key part of the ongoing battle against COVID-19, a place where they could develop multiple vaccine candidates, ready to be rapidly manufactured in response to new variants.

What were the challenges you were seeing for RNA therapeutic manufacturers and how are you helping to address some of these? 

Access to mRNA-LNP R&D and GMP manufacturing expertise with the people and facilities in a single location to support early-stage product development is a challenge for SME’s and academics developing breakthrough medicines in this space.

Post-pandemic, as life starts to return to normal, CPI now has a multi-million-pound facility, one of the largest pools of RNA knowledge in the UK, and the equipment needed to deliver this new technology.  

Now known as the RNA Centre of Excellence, we have a facility that forms the backbone of the UK’s innovation infrastructure in RNA vaccines and therapies. This is the only open-access UK-based centre currently able to develop and manufacture lipid nanoparticle encapsulated messenger and self-amplifying RNA vaccines and therapies ready for use in early-phase clinical trials. It is this legacy that can be exploited in many ways for the benefit of people around the world. 

How does RNA work?

Essentially the principle of mRNA vaccines is, that by providing the gene of a pathogenic protein, the antigen if you like, in the form of messenger RNA to human cells, the cells themselves can then synthesise that antigenic protein to generate the immune response that protects the individual from the actual pathogen.

The antigenic protein is then released or is both displayed on the surface cell and acts as the antigen to stimulate the immune response and provide the vaccination and the protective effect.

So, mRNA vaccines work not by injecting the antigen itself, but by supplying the mRNA that codes for the antigen allowing the cells to do the hard work of making it themselves.

How will RNA be used? 

There are several key drivers for that explosion of activity and interest in RNA vaccines.

• Firstly, it is that they can be produced very quickly, so ideal for healthcare emergencies. They are also easy to make in high yields using platform processes. This means we can do a rapid response and respond to new or mutating viruses.
• We use the body to create the antigen itself so very small doses are required of mRNA compared to other vaccines.
• The relatively low immunogenicity as no viral vector to elicit an immune response.
• RNA vaccines do their work in the cytoplasm so there is no need for them to enter the nucleus like DNA vaccines. This also means that they are non-integrating; so they will not be integrated into the host genome; so there is no risk of infection or mutagenesis. RNA will be degraded by the normal cellular processes in cytoplasm so there is no long-lasting effects.
• Finally, variation in structure and delivery methods chosen can impact the half-life of mRNA which might be important for specific indications.

In summary there a lot of good reasons why people are considering mRNA as a treatment modality. 

What are the challenges? 

Companies are facing challenges around:

• Joined up manufacturing of mRNA vaccines in a single location – including mRNA production but also the lipid nanoparticle encapsulation.
• Access to GMP manufacturing for clinical trial evaluation – at the right scale (often small scale) and at fast pace.
• Access to R&D expertise to evaluate, develop, scale-up, and fully develop commercial processes for mRNA-LNP products.
• Access to R&D expertise for biophysical and biochemical characterisation and quality control of mRNA-LNP products.
• Access to a trained and experienced workforce that can work in both the R&D and GMP manufacturing environments.

eXmoor case study

What does the company do?

eXmoor is a one-stop cell and gene therapy CDMO (contract development and manufacturing organisation), accelerating the manufacturing journey from research to patients. The company has grown over the last 19 years from supporting clients through consultancy for capital and technical projects and now offers in-house process development and GMP manufacturing. Phase 1 of eXmoor’s new purpose-built 65,000 ft² facility includes four clean rooms, a fill and finish suite and a range of laboratories for process development, analytical development and quality control. 

In August 2023 eXmoor formed a global partnership with Kincell Bio, a US-based CDMO that seamlessly brings together both companies’ capabilities, giving customers the opportunity to access a global development and manufacturing footprint in combination with a boutique customer-focused offering.

How will RNA be used?

The COVID-19 pandemic highlighted the significant benefits of mRNA-based vaccines, helping to accelerate the applications of this technology beyond just infectious diseases to other areas such as enzyme replacement and oncology. This has resulted in increased demand for mRNA manufacturing capacity. 

What are the challenges?

mRNA manufacture has unique challenges, and some learning is better transferred from skills developed in cell and gene therapy manufacture rather than classic biologics. eXmoor is uniquely positioned to help with additional capacity and specialist expertise.

Normally, as the scale of a biomanufacturing process increases the price per dose is reduced. But with RNA, driven by the high raw material costs, scale-up results in significant financial risk but only a limited reduction in the price per dose. Vaccine manufacture requires larger batch sizes so process development is likely to focus on efficient use and cost reduction of raw material, as this will have the biggest impact on the price per dose. But not all applications of mRNA need large-scale manufacture and are more sensitive to price per batch than price per dose. 

The high raw material cost used to manufacture mRNA significantly impacts the distribution of costs when working at larger IVT volume as shown below.

What does the future look like for eXmoor?

To support the additional mRNA manufacturing requirements, eXmoor is expanding its portfolio to include an end-to-end service for the manufacture of early-stage mRNA products and therapeutics focusing on flexible scale, rapid turn around and responsive material supply. 

eXmoor’s approach is designed to meet the needs of customers requiring smaller batch sizes and will target IVT scales of up to 0.5l. 

Touchlight case study

What does the company do?

Touchlight is a UK-based contract development and manufacturing organisation (CDMO) and has developed a novel, enzymatically produced DNA vector, known as doggybone DNA or dbDNA™, which is disrupting the decades-old technology of fermentation-based DNA manufacture. With Touchlight’s established low-footprint manufacturing facilities, they produce GMP-grade DNA API and starting materials for viral therapies and RNA therapeutics from the milligram to multigram scale. Extensive investment in facilities has positioned Touchlight as one of the world’s largest DNA manufacturers by capacity, making Touchlight the ideal provider to meet the demands of the rapidly growing advanced therapy market.  

From discovery to GMP, they can offer up to 5 g dbDNA™ per batch in 5 to 6 weeks. Today, dbDNA™ is being implemented by Touchlight’s clients across the genetic medicine spectrum. Within the mRNA space, dbDNA is being used as a critical starting material in multiple clinical trials in the US.

How does the technology work?

dbDNA™ is an enzymatically produced, linear, double stranded and covalently closed minimal DNA vector with no bacterial backbone sequences, and no origins of replication or antibiotic resistance genes. dbDNA™ is first amplified from minute amounts of DNA template by the high-fidelity Phi29 polymerase. The resulting concatemers are linearised and covalently closed at the ends by a proprietary TelN mediated procedure, allowing for removal of backbone and accessory sequences through concerted action of sequence-specific endonuclease and exonuclease. The dbDNA™ is then purified applying state-of-the-art chromatography and filtration methods and controlled for safety and purity before release with established quality control methods (QC). dbDNA™ can encode long, complex, and unstable DNA sequences such as structured regions, repeats or homopolymer stretches like polyA, which otherwise would cause toxicity in E. coli cultures, and rendering unpredictability to fermentation-based DNA manufacture.

How is dbDNA used for RNA manufacture?

De-ending dbDNA™ through restriction digestion results in zdbDNA™, which acts as starting material for run-off in vitro transcription (IVT). The minimal sequence and its enzymatic manufacturing enable consistent production of large quantities of highly pure sequences with the dbDNA™ technology. Together with the potential to use lower template amounts in IVT, dbDNA™ constitutes an optimal starting material for GMP manufacturing of diverse RNA modalities.

What are the challenges?

The recent pandemic has highlighted the need for rapid, scalable, and seamlessly transferable vaccine production. Although RNA vaccines have demonstrated potential to combat unprecedented infectious disease, rapid provision of clinical trial material, and subsequent commercial supply and resupply remain challenging. A critical bottleneck for rapid onset of clinical trials as well as for resilient supply of commercial RNA products is the production of plasmid DNA, which often fails to provide the starting material for manufacture of mRNA and the rising self-amplifying or circular RNA modalities reproducibly and consistently. Touchlight is alleviating that bottleneck through enzymatic production of dbDNA™, and delivers at unparalleled speed, reproducibility, scale, and purity. Touchlight relies on a highly skilled and experienced team of RnD, GMP, QC, regulatory and commercial experts to support customer-specific demands and to enable on-time provision of high-quality templates for diverse RNA modalities.

New England Biolabs case study

What does the company do?

Founded in 1974, New England Biolabs (NEB) is a leading company developing and manufacturing enzymes and reagents for life sciences research. They specialise in products crucial for mRNA production, offering solutions for both research and commercial-scale manufacturing. Their offerings span the entire workflow, ensuring consistent high-quality mRNA at all stages.

How does NEB’s technology work?

NEB provides a comprehensive suite of enzymes and kits for in-vitro transcription (IVT), the process of creating mRNA from a DNA template. Their products include:

• T7 RNA polymerase: The workhorse enzyme for mRNA synthesis.
• NTPs: Building blocks for constructing the mRNA molecule.
• DNAse I: Eliminates the DNA template after transcription.
• Murine RNase inhibitor: Protects newly synthesised mRNA from degradation.
• Capping enzymes: Add a crucial modification (cap) to the mRNA for stability and translation efficiency.
• mRNA Cap 2’-O-Methyltransferase: Further refines the cap structure for optimal function.
• Restriction enzymes (optional): Linearize plasmids for efficient IVT.

NEB offers both research-use-only and GMP-grade enzymes, catering to various development stages. Additionally, they provide large-scale production capabilities to meet the growing demand for mRNA therapeutics.

What are the challenges?

NEB understands the challenges of scaling up mRNA production. They’ve addressed this by:

• Investing in infrastructure: Increased fermentation capacity allows them to produce enzymes in bulk quantities.
• Early client engagement: Working with clients early on ensures a smooth transition from research to large-scale manufacturing.
• Diverse product formats: Offering enzymes in various sizes caters to different research and production needs.

NEB emphasises the importance of choosing a reliable raw material supplier for mRNA production. They highlight their strengths in this area:

• Quality management: Certified facilities and rigorous quality control processes ensure consistent product performance.
• Technical expertise: NEB scientists are well-versed in mRNA workflows and can provide valuable support.
• Transparency: NEB encourages client visits to their facilities and welcomes audits, demonstrating confidence in their manufacturing practices.
• Future-proof compliance: NEB actively monitors regulatory changes and ensures their products remain compliant.

Questions therapy developers and manufacturers should ask their suppliers

In addition to the points mentioned above, here are some key questions therapy developers and manufacturers should ask their raw material suppliers:

• Can the supplier guarantee consistent batch-to-batch performance of their enzymes and reagents?
• Does the supplier offer a wide range of products to support the entire mRNA production workflow?
• Can the supplier scale up production to meet the demands of commercial-scale manufacturing?
• Does the supplier have a proven track record of supplying high-quality products to the mRNA therapeutics industry?
• Is the supplier willing to collaborate with clients to develop custom solutions?
• Does the supplier offer technical support to help clients troubleshoot issues?

By considering these questions, therapy developers and manufacturers can select a reliable raw material supplier that can help them bring their mRNA therapies to market.

What does the future look like for NEB?

NEB is committed to supporting the advancement of mRNA technology. Their focus areas include:

• Developing new enzymes and reagents: Continuously improving tools for mRNA production.
• Personalised medicine: Providing solutions for small-scale batches tailored to specific needs.
• Workflow simplification: Streamlining processes to save time and resources for researchers.
• Lyophilisation technology: Offering enzymes and mixes stable at ambient temperature, simplifying logistics.

NEB is well-positioned to play a key role in the ongoing mRNA revolution by providing scientists with the tools and expertise needed to develop next-generation therapies.

Note: The branding term “GMP-grade” has been clarified to avoid confusion with official GMP regulations.

TriLink case study

Kate Broderick

Chief Innovation Officer, TriLink

It is my great pleasure to provide a foreword to the introduction of TriLink Biotechnologies. As a proud Scot/Brit who received my degree and PhD from the University of Glasgow prior to moving to the States, the critical work that BIA does to connect developers in the US with innovation in the UK is vital.

At TriLink, we believe that the future is RNA. The remarkable efficacy of the approved COVID-19 mRNA-based vaccines was only the start of the RNA-based medicine revolution. As an innovative CDMO, TriLink is honoured to support RNA developers as they bring their paradigm-shirting therapeutics and vaccines to patients. Our 20+ years of experience in the RNA field allow us to provide world-class expertise in the products and services critical for nucleic acid breakthroughs.”

What does the company do?

TriLink BioTechnologies, part of Maravai LifeSciences, is a contract development and manufacturing organisation (CDMO) for the synthesis of nucleic acids, nucleotide triphosphates (NTPs), and mRNA capping analogs. TriLink has scale-up expertise and unique mRNA, oligonucleotide & template sourcing capabilities for companies focused on therapeutic, vaccine, diagnostic and biopharmaceutical breakthroughs. TriLink has been working in mRNA for over 20 years and was one of the first to provide GMP mRNA manufacturing. To date, TriLink has generated over 100 batches of GMP mRNA to be used in clinical trials. Our CleanCap® technology is in three of the four approved COVID-19 vaccines being used globally today. It was included in the capping technology used in the Pfizer BioNTech COVID-19 vaccine.

How does the technology work? 

mRNA capping plays a critical role in the success of mRNA vaccines and therapeutics. The cap is a specialised structure placed at the beginning of the mRNA molecule that acts to prevent premature degradation, avoids triggering an unwanted immune response and ensures the mRNA reaches its intended destination and generates the desired protein product. CleanCap® mRNA capping technology offers a streamlined approach as the capping and mRNA synthesis occur in one step, referred to as co-transcriptional capping. The analog is extended by the RNA polymerase during In Vitro Transcription (IVT), enabling high capping efficiency of >95% in a one-pot reaction. Crude mRNA from the co-transcriptional CleanCap® reactions can be purified by industry standard methods including oligo dT affinity chromatography.

CleanCap® AG, AG 3′ OMe, M6, and CleanCap® AU reagents are shown above. All cap analogs result in Cap1 mRNA, the most abundant cap structure in eukaryotic cells. CleanCap® AU was developed for self-amplifying RNA to maintain the natural 5’ UTR sequence of alphavirus vectors used in this technology, while the AG series of CleanCaps® are available for traditional linear mRNA. The +1 base, or what becomes the first base in the transcribed mRNA is an A, and the second base is a G (for 3 of the 4 analogs) for use on matched DNA templates. The +2 base is an important part of the cap analog as this allows for RNA polymerase extension from the 3’ hydroxyl position of the unmodified ribose.

What advantages does CleanCap® technology offer?

There are different approaches to capping. Clean cap uses co-transcriptional capping as discussed above, whereas enzymatic capping utilises enzymes to add the cap after the mRNA has been made, resulting in more hands-on time to produce final desired mRNA. The CleanCap® M6 reagent is the newest cap analog available from TriLink. CleanCap® M6 offers a modified Cap 1 structure that is currently only achievable through cap analogs. The protein expression levels of M6 capped mRNA can be measured using an mRNA that encodes for a bioluminescent protein that emits light. The luciferase activity correlates with protein expression levels. The luciferase mRNA capped by the CleanCap® M6 reagent outperforms enzymatically capped Cap1 luciferase mRNA in mice as shown below. With the increased potency offered by this latest cap analog it may be possible to decrease mRNA doses in certain therapeutic applications.

Performance of FLuc mRNA in an LNP-formulated, 1 mg/kg by tail vein delivery into mice. Luciferase activity (photons per second) was measured after luciferin injection from 3-48h for total AUC graphed here. *** p < 0.001, two-tailed T test. Error bars are standard error of mean. n = 9/group.

How does TriLink support the community?

TriLink, through Maravai, is a member of the Alliance for mRNA Medicines (AMM), an organisation dedicated to advancing and advocating for mRNA and next-generation encoding RNA therapeutics and vaccines for the benefit of patients, public health, and society. AMM’s mission is to propel the future of mRNA medicine, improve patients’ lives, and advance scientific knowledge by convening and empowering mRNA industry leaders, innovators, scientists, and other key stakeholders.

TriLink has supported countless custom development and R&D collaboration projects. For example, during the height of the COVID-19 pandemic in 2020 the TriLink team pivoted resources to support a critical troubleshooting project for the manufacturing process of Imperial College London’s saRNA vaccine prior to a tech transfer back to the UK. Additionally, the Science and Innovation Office led by Dr. Kate Broderick is always available to discuss novel academic collaborations (scan QR code if interested).