Category: mRNA

Predictions for double-digit growth of the mRNA market have many small and emerging biotechs joining big pharma companies in pursuing the development of novel mRNA-based therapeutics and vaccines. The complex, multistep process involved in mRNA–LNP manufacture poses a host of novel challenges, particularly as processes are scaled. The risks involved with coordinating raw material supply, scaling, and aligning steps across multiple vendors can be mitigated by partnering with an experienced CDMO with end-to-end capabilities from cell banking to fill/finish of LNP-formulated mRNA products.

mRNA Promises Still Abound

The value of mRNA technology to human society was properly recognized in early October 2023 with the awarding of the Nobel Prize in Physiology or Medicine to Professors Katalin Kariko, Ph.D.,  and Drew Weissman, M.D., Ph.D., for the contributions each made to its development during their time as researchers at the University of Pennsylvania. Of course, that award was far from the only public recognition of the importance of their work, with millions worldwide having been protected by the mRNA-based vaccines developed in response to the COVID-19 pandemic.

Going forward, the market value of the mRNA vaccines and therapeutics is estimated to continue to grow rapidly for the foreseeable future. This market is estimated by several research firms
to be approximately $40–50 billion in 2022/2023 and to be expanding at a compound annual growth rate between 13% and 17%. Notably, the therapeutics sector is expected to be the major driver of further growth as demand for COVID-19 vaccines levels off.

Maximum Growth Requires Surmounting Manufacturing Hurdles

Unlike traditional biologics, which are produced via microbial fermentation or mammalian cell culture, mRNA production does not require the use of living cells and is therefore simpler overall. However, that does not mean that consistent manufacture of high-quality mRNA is easy. Lab-scale kits cannot be practically implemented at commercial scale, process control can be challenging at large scale, and batch-based processes involve many steps for which there has yet to be standardization implemented across the industry. Furthermore, mRNA instability poses significant challenges. Raw material variability and the lack of fit-for-purpose, single-use manufacturing equipment exacerbate the growing pains in this young sector.

Wacker Biotech supports clients throughout the entire journey from pDNA to the LNP formulated mRNA product

The sensitivity of mRNA molecules to temperature, shear, and degradation by ubiquitous enzymes creates the need for extensive process controls. Furthermore, processes for the generation of final products in the form of lipid nanoparticle (LNP) encapsulation of the mRNA active, which is necessary to improve stability and facilitate delivery into appropriate tissues, typically involve the use of proprietary lipids and large quantities of solvents.

Both biopharma companies and academic groups are working on solutions to these challenges to enable more robust, scalable, and cost-effective mRNA manufacturing processes. In the meantime, many developers of mRNA therapeutics and vaccines are turning to outsourcing partners with the expertise and specialized capabilities needed to produce mRNA drug substances and mRNA–LNP drug products.

Complex, Multistep Process

The production of mRNA begins with transcribing the genetic code from linearized plasmid DNA (pDNA) into mRNA. While some manufacturers create pDNA through bacterial fermentation and linearization as initial steps, many acquire pDNA from external sources. The pDNA template carries a DNA-dependent RNA polymerase promoter and the mRNA construct’s corresponding sequence. In vitro transcription (IVT), a cell-free process employing RNA polymerases derived from bacteriophages and chemically modified ribonucleoside triphosphate molecules (rNTPs), then transforms pDNA into mRNA, enhancing stability with a 5’ cap and 3’ poly-(A) tail. Downstream purification includes tangential-flow filtration (TFF) and chromatography to remove immunogenic double-stranded RNA (dsRNA), residual DNA template, RNA polymerase, and elemental impurities, followed by mRNA encapsulation in lipid nanoparticles (LNPs) using various lipid types and ratios. LNPs are formed via rapid mixing of lipid solutions with mRNA, and the final product is prepared in the desired buffer solution and concentrated using TFF.

mRNA molecule

Sequence- and Scale-Dependent Bioprocessing Considerations

The specific steps in mRNA production vary from manufacturer to manufacturer and as a function of the genetic sequence, target application, dosage, and scale. In addition, the quantity and dosage level for mRNA products vary significantly depending on the application, from large-scale vaccines to fight global pandemics to very-small-scale personalized cancer therapies. Scalability — whether up, down, or out — is therefore essential, regardless of the manufacturing strategy. Processes must provide high yields of high-quality mRNA products cost-effectively over a range from micrograms to kilograms.

Although process parameters for the IVT step will need to be adjusted depending on the specific sequence, this step of the mRNA–LNP drug product production process is fairly platformizable. Downstream purification of the mRNA drug substance can also differ, depending on the nature of the mRNA. The optimum unit operation can also vary with scale, so in some cases scaling up and down is not simply a matter of changing equipment sizes but using different equipment entirely.

Also, the pDNA and LNP manufacturing need to be adjusted depending on the product; the size and other properties of the plasmid determine the optimum upstream process parameters for pDNA production, while the mRNA sequence dictates the choice of lipids, lipid ratios, and mixing parameters. Consequently, extensive process development and optimization is often needed for each of the three major steps of mRNA manufacturing and formulation.

The instability of mRNA, including its sensitivity to temperature, shear, and ubiquitous RNase enzymes, must be managed not only during production but also during analysis and storage/distribution. Minimizing processing and handling times and freeze-thaw steps is crucial, as are monitoring and control of all process steps; all become more challenging when large process volumes are involved. Rapid analytical methods suitable for product characterization, quality control release, and stability testing, are also necessary. One way to help minimize overall production times and handling of mRNA is to establish end-to-end manufacturing capabilities from pDNA through mRNA–LNP fill/finish.

Sourcing difficulties, meanwhile, create additional challenges for scaling mRNA–LNP production. Regulatory authorities today expect that not only critical raw materials but also important ancillary materials meet GMP requirements. Access to a reliable supply of high-quality and GMP-compliant raw materials for mRNA production, including pDNA and the specialized enzymes and lipids required for IVT and LNP formation, are of particular note. The limited number of suppliers that can reliably provide high-quality and GMP-grade specialty enzymes and lipids in large quantities presents cost and availability issues.

Implementing Effective LNP Processes

Different mixing technologies can be leveraged to produce LNPs: microfluidics, such as the continuous technology developed by Precision NanoSystems, and more traditional mixing in classical mixers. Both are widely used depending on the needs and preferences of each drug developer. Development of effective processes using both approaches involves trial and error to some extent, however, because full and deep knowledge of the LNP formation process remains lacking.

Management of large quantities of flammable solvents is a separate challenge associated with current LNP processes. Such materials are not found in typical biopharmaceutical manufacturing facilities and require the implementation of special safety measures. Containment infrastructure, solvent waste disposal, and operator training are just three additional items that add cost and complexity to LNP manufacturing. Some companies elect to run multiple smaller batches rather than install the costly infrastructure required to perform larger-scale runs. In addition, because single-use equipment is not compatible with most organic solvents used in LNP processes, stainless-steel equipment, with necessary cleaning and sterilization systems, must be employed.

Wacker Biotech Offers End-to-End mRNA CapabilitiesWacker Biotech is an established and reliable CDMO with extensive experience in the development and manufacture of many different types of biologics employing both stainless-steel bioreactors and single-use technologies, including conventional recombinant proteins, antibody fragments, fusion proteins, scaffolds/binding proteins, cytokines/interleukins, live biotherapeutic products (LBPs), vaccines, enzymes, peptides, and plasmid DNA, among others.This versatile development and production experience also gives Wacker Biotech the ability to effectively respond to customer needs in the mRNA space. In addition, the Wacker Biotech team has experience scaling these diverse types of processes, developing the necessary analytical methods for process monitoring and product release, and implementing the processes in the GMP production environment.Before the emergence of the COVID-19 pandemic, Wacker Biotech had already begun investigating in mRNA-specific technology. Indeed, during the pandemic, mRNA COVID-19 vaccine developer CureVac selected Wacker Biotech over other CDMOs to manufacture its candidate because it deemed WACKER best positioned to quickly meet its production needs. The company met expectations. In an environment with difficult material supply challenges, Wacker Biotech transferred an mRNA production process for that vaccine from the lab to commercial scale.In Germany, WACKER has secured government funding to construct a specialized facility at the Halle site, designed to enhance pandemic preparedness. This facility, set to be operational by April 2024, is uniquely structured with four production lines, accommodating varying capacity needs for pDNA, mRNA drug substance, final mRNA–LNP drug product manufacturing, and fill/finish operations. Wacker Biotech is developing these capabilities with agility in mind. The facility’s design draws on Wacker Biotech’s expertise to optimizes efficiency and the flexibility to adapt to evolving regulatory requirements and the dynamic landscape of novel modalities. Wacker Biotech’s exclusive selection as the German government’s sole CDMO partner for this pivotal pandemic preparedness initiative underscores their commitment to addressing healthcare challenges swiftly and effectively.Proprietary platform processes are being developed by Wacker Biotech for both pDNA and mRNA. A primary advantage is greater understanding, which facilitates better predictability concerning how different sequences will behave. Using standard unit operations also enables the stockpiling of important raw materials. For instance, Wacker Biotech has developed standard filter setups, bags, and buffer preparations, which are all maintained in ample quantities in the company’s warehouses.Wacker Biotech has also prioritized developing expertise in a variety of LNP process types that operate at numerous different volumes. Having capabilities in both microfluidic and traditional mixing technologies creates opportunities for a CDMO to meet the expectations of a broader range of mRNA developers. In the meantime, the company has partnered with CordenPharma, a leading expert in the production of lipids for LNP formulation. Integrating CordenPharma’s lipid production expertise and raw material supply capabilities with Wacker Biotech’s mRNA therapy development and manufacturing leadership delivers unprecedented capabilities.These efforts are complemented by Wacker Biotech’s R&D expertise. Its laboratories are outfitted with small-scale, high-throughput screening capabilities that enable thorough exploration of many different process parameters and thus greater process understanding at a reasonable cost. The process scale-up team can then confirm process performance at medium scale before transfer to production.With expertise in pDNA, mRNA drug substance, and mRNA–LNP drug product manufacturing, including fill/finish operations, Wacker Biotech can provide comprehensive, end-to-end services in support of mRNA vaccine and therapy development and production. Plasmid DNA development and production (high-quality and/or GMP mRNA) takes place in San Diego, with mRNA drug substance and mRNA–LNP drug product processes developed in Amsterdam and GMP clinical and commercial production taking place both in Amsterdam and Halle, where four parallel production lines are available.As the pharmaceutical landscape evolves rapidly, Wacker Biotech continues to fortify its position at the forefront of mRNA manufacturing. With its strategic investments, adaptable facilities, and an unwavering commitment to meet both present and future demands, WACKER stands ready to address both unforeseen global health challenges and the specific needs of its clients. The company’s proactive approach to expanding and diversifying its capabilities through platform innovation and strain and formulation development, among other activities, reaffirms its dedication to innovation and excellence in a highly competitive space. As the world looks toward the future of medicine, Wacker Biotech is poised to be an instrumental partner in the next generation of healthcare solutions.

Addressing Coordination Challenges

Given the large number of manufacturing steps involved in the production of mRNA products through fill/finish and final packaging, it is not surprising that manufacturers lacking appropriate in-house capabilities frequently end up working with multiple service providers. That adds time, cost, and risk to the overall process, as well as greater supplier management requirements.

These challenges can be greatly reduced by working with a CDMO that provides an end-to-end, integrated offering starting with plasmid design, cell banking, and plasmid production and continuing through fill/finish of mRNA–LNP formulated products. Planning and oversight can be greatly simplified, with each step in the process well aligned. Even when transfer from one site to another is still required, those transfers occur within the same company with much reduced delays, cost, and risk, particularly if all of the sites operate under the same management, quality, and regulatory systems, have the documentation styles and review expectations, and approach raw material procurement in the same manner. Development, analytical, and production teams can also meet regularly to ensure that all activities are completely and continuously aligned across the entire value chain.

Wacker Biotech’s mRNA Competence Center in Halle, Germany (expected to be operational by April 2024)

Finding such a CDMO can be a challenge, however, because very few have experience in large-scale manufacture of mRNA drug substances and mRNA–LNP drug products. Those that do have typically focused on one of the steps — pDNA manufacture, mRNA production, mRNA–LNP formation, or fill/finish operations. Drug developers must therefore coordinate the efforts of a multitude of suppliers and manage longer timelines, greater risk, and higher costs. Effective process improvements are inhibited, and the instability of mRNA magnifies the challenges.

A Need for CDMO Flexibility

Flexibility is in fact crucial for mRNA drug developers and hence for CDMOs offering support for the development and manufacture of mRNA products. The variability in mRNA processes is further complicated by a complex intellectual property (IP) landscape, which drives each developer of mRNA vaccines and therapeutics to seek different methods to skirt existing patents, often in downstream purification techniques (e.g., preparative high-performance chromatography, ethanol precipitation, clarification chromatography) or in the LNP production step.  If the CDMO offers a (platform) production process, the CDMO becomes responsible of making sure it has the freedom to operate within this landscape.

It is also worth noting that regulatory requirements for mRNA drug substances and drug products have not yet been  fully established. Both the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMA) are still learning as candidates move through the development and approval process, and further guidance is expected. More changes are anticipated with regard to analytics compared with processing.

Originally published on PharmasAlmanac.com on April 8, 2024.

The process for manufacturing messenger RNA (mRNA) vaccines and therapeutics comprises two main steps: production of the mRNA payload, which is translated inside the body into proteins that produce antibodies or impact disease mechanisms in some way; and production of the lipid nanoparticles (LNPs) encapsulating the mRNA to provide protection and facilitate delivery into cells. The unique nuances of mRNA production — which can impact safety, efficacy, quality, and manufacturability — and the inherent fragility of mRNA molecules drive the need for a host of specialized capabilities and expertise. Strict ultra-cold chain processes and protocols are necessary to maintain material viability and quality. An outsourcing partner like Samsung Biologics that can provide true end-to-end manufacturing support combined with effective material sourcing and distribution management strategies can accelerate project timelines while reducing risks.

Challenges to mRNA Production and Supply

Unlike most typical recombinant protein and antibody drug substances, mRNA is inherently fragile and prone to degradation. The phosphodiester bonds in the backbones of single-stranded mRNA molecules are cleaved via transesterification by 2′-hydroxyl groups, a reaction that can be catalyzed by water. Ribonucleases (RNases), enzymes widely present in the environment, and other ribozymes also catalyze phosphodiester bond cleavage.

During manufacturing, RNase contamination control is crucial to ensure the manufacture of high-quality mRNA products. Gamma-irradiated, single-use equipment is strongly preferred to avoid possible contact with problematic enzymes. Samples and mRNA products must be stored at low temperature (–20 to –70 °C) to suppress degradation reactivity.

Encapsulated mRNA payloads are also typically unstable at room temperature. The temperature during LNP manufacturing must be closely monitored and controlled. In addition, carefully controlled and dedicated processes for freezing and thawing must be developed, as the timeline for both can affect the final particle size and encapsulation efficiency. The temperature-dependent sensitivity of mRNA–LNPs must also be addressed via controlled low-temperature storage and handling.

Sourcing of raw materials and single-use consumables, meanwhile, requires careful approaches and strategic relationships with suppliers. Specialty lipids and tailor-made plasmid DNA are high-value raw materials; suppliers must be selected under strict quality standards, and business transactions must be handled appropriately. Unit price, delivery, and quality dispute terms should be closely evaluated when establishing supplier agreements.

Some critical raw materials also require strict storage and handling conditions to ensure that their high quality is preserved. As a result, flexible temperature-controlled storage areas covering a range of temperatures are required to ensure that not only mRNA drug substances and mRNA–LNP drug products, but also other key raw materials, can be securely stored while maintaining their quality.

The COVID-19 pandemic created additional supply challenges. Soaring demand for both plasmid DNA and other raw materials (e.g., lipids, capping reagents, etc.) required for mRNA production, as well as components for single-use systems, led to shortages that persist today. For instance, when the pandemic emerged, only few suppliers produced GMP-grade plasmids at large scale. Consumables were, meanwhile, increasingly used at manufacturing scale not just for mRNA vaccines but also for COVID-19 therapeutics and many non-COVID biologics.

In the face of this skyrocketing demand, vendors responded quickly to expand existing production capacity and build new manufacturing facilities, albeit not quickly enough to meet demand and avoid inevitable delays for drug manufacturers. Much of that new capacity has recently come onstream or will in 2023, and supply issues are therefore somewhat alleviated today and should continue to improve going forward.

Global Supply Chain Shifts

COVID-19 impacted all aspects of the global pharmaceutical supply chain due to lockdowns, raw material supply shortages, lower shipping container availability, route closures, reduced personnel, less frequent commercial air traffic, and many other factors. Prioritization (out of necessity) of the COVID-19 therapeutics and vaccines ahead of existing APIs and drug products made it even more challenging for biopharma companies and their outsourcing partners to secure inventory, even for raw materials. Enhanced collaboration, proactive monitoring technologies, and implementation of innovative supply chain management strategies helped streamline sourcing and reduce risks.

The effects of this supply chain crisis continue to linger. Production scheduling remains challenging because there is often no assurance as to when raw materials will be available. With few qualified raw material suppliers offering products suitable for GMP manufacturing, demand for GMP-grade, mRNA-specific raw materials has increased dramatically, as have lead times. A lack of specific raw materials can force the use of substitutes/alternatives, but the risks of doing so — when possible — must be clearly understood before manufacturing is initiated. Use of unqualified raw materials can cause risks of degradation if appropriate low-temperature storage space is insufficient.

Difficulty accessing single-use consumables owing to high demand and limited supply also continues to affect production scheduling. Lead times for these materials have increased more than twofold in some cases. The heavy reliance of mRNA manufacturing on single-use equipment for contamination control may well expand the need for localized production of single-use technologies to minimize supply shortages and to enable suppliers to maximize business opportunities.

Cold chain management has become strategically imperative for the industry in general, with mRNA-specific logistic route management becoming truly essential for mRNA manufacturers. Effective management of cold-chain logistics for mRNA vaccines and therapeutics requires precise coordination from start to finish, including temperature monitoring, real-time tracking for traceability, and well-trained and skilled logistics personnel, to ensure retention of efficacy.

Supporting Continued mRNA Production

Manufacturers of conventional biologics (recombinant proteins and monoclonal antibodies) have cold chain capabilities, but they are not typically set up to support the extreme temperatures often required for mRNA–LNP products. In order to adapt to global supply chain shifts, contract development and manufacturing organizations (CDMOs) have thus expanded their cold chain and fill/finish offerings to address the additional requirements for new modalities, such as mRNA.

Multiple types of storage capabilities, each with appropriate standard operating procedures, have become essential. Notably, mRNA–LNP products must be carefully frozen and thawed in blast freezers that provide storage at temperatures from –20 to –70 °C, a process typically achieved using control-rate freezers, which require management and operational resources to be effective.

Flexibility in primary packaging is also important for mRNA product developers, as is validation of drug products. If all handling of the mRNA–LNP product takes place on site with little likelihood of temperature variations that, if they do occur, can be more easily controlled, static validations are suitable. Off-site drug product–handling activities (e.g., transport) require dynamic validations, because greater variation in conditions, including temperature shifts, are expected.

End-to-End CDMOs Offer the Best Solution

CDMOs that offer end-to-end support for the entire mRNA workflow can more easily overcome some of these challenges due to their experience across the entire spectrum (from sourcing raw materials, reagents, and single-use equipment to implementing and monitoring temperature controls across each phase of development and distribution). The supply chain is also greatly streamlined by eliminating the need to work with numerous vendors, saving both time and cost.

Equally important, risks are significantly reduced. Having mRNA drug substance manufactured at one site, then transferred to another for encapsulation, and a third for fill/finish introduces opportunities for product loss given the fragile nature of mRNA. Every time the material experiences high-temperature excursions, it is possible that quality, efficacy, and safety can be negatively impacted.

Ideally, mRNA drug substance should be either transferred directly to the LNP formulation step or used immediately after thawing and stabilization to achieve maximum yield and purity of the encapsulated payload. With mRNA drug substance manufacturing, mRNA–LNP drug product production, and final fill/finish operations located within the same facility and supported by on-site analytical services — as is the case at Samsung Biologics — drug and vaccine companies can have much greater confidence that their products will be safely and securely manufactured with the highest quality and efficacy while avoiding common supply chain challenges.

Samsung Biologics’ clients also benefit from its ability to achieve biologic product approvals from global regulatory agencies including the U.S. FDA and the EMA, among others. The company has also repeatedly demonstrated capabilities with respect to validated processes for the manufacturing, packaging, and shipping of products that require cold-chain distribution within a wide range of temperature-controlled environments, as well as storage and handling of raw materials that require cold chain management. The depth of knowledge and expertise gained from this experience has direct applicability to mRNA–LNP product shipping and supply.

End-to-end support also facilitates scale-up of mRNA drug substance and drug product manufacturing, which will become increasingly important as the many candidate vaccines and therapeutics in the clinical pipeline advance to later development stages and ultimately to commercialization. The ability to perform all activities from lab to commercial scale at one facility eliminates unnecessary time loss, cost, and risks associated with the transfer of materials from one site to another.

Agile and Transparent Communication Simplify Sourcing

Samsung Biologics and its clients have benefited from an approach to sourcing that emphasizes ongoing open communications with both suppliers and customers. For all activities, including sourcing, client satisfaction is a constant driver.

Each client works with a dedicated team of technical experts that includes representatives from procurement. At weekly supply meetings, project updates are shared, followed by a brainstorming session to lead to the best-case scenario and establish specific action items. In addition, client product forecasts are converted to raw material forecasts, which are then quickly shared with vendors to enable sufficient supply at the right time.

Samsung Biologics also applies a concurrent engineering concept to tech-transfer projects, with dedicated experts remaining engaged with a project throughout its lifetime. This approach ensures that the people with the most knowledge about the process are involved in technical discussions with the client and can help to ensure on-time delivery by resolving any quality or other issues that arise. In addition, the supply chain management team communicates directly with the technical and manufacturing teams in order to overcome challenging supply issues. The end result: tech-transfer projects (both conventional biologics and mRNA) are typically completed within six months, faster than the industry average.

 In the case of mRNA production, Samsung experts focus on eliminating bottlenecks involving single-use components from customized equipment assemblies by working closely with technical experts at external vendors. Early-stage joint communication also accelerates the forecast and ordering process, enabling the completion of tech transfer within six months. Combined, these efforts result in significantly increased raw material readiness.

As a top-10 global CDMO with extensive mAb manufacturing experience, Samsung Biologics has also established close relationships and strong ties with many global suppliers. These relationships have been leveraged to expedite access to important raw materials and single-use components for mRNA production.

Building Local Supply Chains to Enhance Security of Raw Material Supply

In addition to a team approach that emphasizes continuous, transparent communication within Samsung Biologics and between suppliers and customers, multi-sourcing has become an important and successful strategy for addressing supply chain challenges. Sourcing raw materials from multiple suppliers helps ensure access to key ingredients. If some of those different suppliers are local, the security of the supply chain is even greater.

This philosophy has led Samsung Biologics to attract major biopharma raw material manufacturers into Songdo, the city in South Korea in which Samsung’s manufacturing site is presently located. This area is rapidly becoming a key hub for the production of important raw materials and single-use components used in biologics manufacturing, including mRNA–LNP products. Sourcing materials coming from manufacturers in Songdo, Samsung Biologics’ headquarters, helps to minimize lead times. As additional raw material producers establish nearby manufacturing sites, stability of supply will be further enhanced, and Samsung Biologics’ internal efficiency will increase even further.

Established Cold Chain Processes

With today’s distribution network becoming increasingly complex at a time when biopharma companies face new challenges, Samsung Biologics ensures on-time delivery of high-quality mRNA–LNP products leveraging effective sourcing strategies and cold chain capabilities.

Strong and deep relationships with raw material suppliers ensure consistent supply of mRNA-specific raw materials and single-use components. Investment in local supply chain infrastructure facilitates collaboration with not just local suppliers but also global partners and regulators. Dedicated manufacturing capacity ensures the production of high-quality mRNA drug substances and mRNA–LNP drug products, and extensive infrastructure for cold chain supply enables storage and handling of mRNA drug substances and mRNA–LNP drug products at temperatures ranging from –20 to –70 °C. Products and samples are packaged with precision for distribution and stored within temperature-regulated and highly controlled areas in compliance with strict quality standards before being released.

Prepared to Manage Future Uncertainties with Newer Modalities Beyond mRNA

Biopharma companies developing new modalities, such as mRNA therapeutics and vaccines, face uncertainties from two directions. On one end, predicting demand for new classes of drugs is difficult. On the other, for mRNA-based candidates in particular, accessing large quantities of high-quality, GMP-grade plasmids and fit-for-purpose, and single-use assemblies is even more difficult. Added to these uncertainties is the general lack of industry knowledge about producing novel drug substances and drug products at large scale.

Samsung Biologics is tackling these uncertainties head on. The company’s development and manufacturing systems have built-in flexibility and agility, enabling rapid responses to changing market dynamics that influence both supply of key raw materials and quality for final drug products. The localized supply approach provides an ecosystem better equipped to respond to sudden crises. As an end-to-end CDMO, Samsung Biologics also integrates typically segmented production operations into consistent processes — all at one site — that allows for greater control and affords high-quality products. The company is also applying its knowledge and expertise of mAb manufacturing where appropriate, to address challenges to the production of new modalities, such as mRNA.

Originally published on PharmasAlmanac.com on March 9, 2023.

Drug development, regardless of the modality, invariably involves some level of risk. As mRNA therapeutics advance, they too will face such risks, including some analogous to those posed by other modalities and some unique to mRNA. Manufacturing processes must be robust, efficient, scalable, and readily monitored and controlled — not an insignificant challenge, particularly given the drive to establish them quickly. BIOVECTRA has explicitly taken steps to reduce the risks associated with mRNA development and manufacturing, including using automation combined with a quality-by-design approach for efficient and effective process optimization, development of in-house analytical methods, and constant monitoring of technology advances and the evolution of regulatory guidance.

mRNA: Highly Promising Opportunities, but not Risk-Free

With the robust efforts across the industry to rapidly and efficiently advance mRNA therapeutics, a broad range of risks must be considered and mitigated. Drug developers and their manufacturing partners are driven to develop robust, efficient, reliable, and scalable manufacturing processes in the context of competitive market pressures. Regulatory guidelines for evaluation of the safety, quality, and efficacy of mRNA therapeutics and vaccines are evolving, with draft guidances likely to become formalized in the near future.

Comprehensive Risk-Management Approach at BIOVECTRA

Risk management at BIOVECTRA begins with a quality-by-design (QbD) approach to ensure deep understanding of critical process parameters (CPPs) and critical product quality attributes (CQAs). Manufacturing processes are then designed based on these principles.

Another component of BIOVECTRA’s risk-management approach involves relying on a team of experts with a broad array of knowledge and experience. In addition to quality assurance, engineering, analytics, process development, and manufacturing, science and technology groups participate in risk assessments, applying their diverse perspectives to ensure the best possible process designs.

A third element of the BIOVECTRA risk-management strategy considers, through project management exercises, what risks are associated with advancing our internal platforms and/or client projects.

Underlying these three aspects of our approach to managing the risk of mRNA manufacturing is the implementation of process automation solutions wherever feasible in the new production facility, including the use of process analytical technologies (PATs) to monitor CPPs.

Balancing Innovation and Risk with Automation

Automation has been proven in other industries to result in more robust processes and to facilitate more streamlined program advancement. For mRNA projects, automation also has the potential to reduce development times, allowing clients to move from preclinical R&D to clinical trials efficiently, which is highly desirable as drug developers race to be the first to market with these next-generation therapies.

Indeed, given that mRNA synthesis typically requires the same process conditions and reagents (e.g., nucleotides, enzymes, DNA substrate) with only the genetic sequence of the DNA varying, it can be straightforward to automate. A bioreactor with inline PAT providing feedback on the temperature, pH, conductivity, and other factors allows careful control of the process. Purification can then be performed via automated filtration followed by chromatography with real-time fraction UV analysis, followed by ultrafiltration, concentration, and buffer exchange.

The pressure for biopharma companies to invest in artificial intelligence, robotics, and digitalization to improve process performance is likely to continue to rise. BIOVECTRA believes that the maturation of the mRNA field (and the pDNA field by association) creates opportunities for leading-edge manufacturers to leverage these technologies, as well as continuous processing. AI can help to predict process outcomes and lead to safer, more efficient, and more productive processes.

One solution we are exploring is the digital twin, in which robotics sensors and AI allow collection of real-time process data that is then used to generate a simulation of that specific process. The simulation is then used to analyze process performance and identify areas for improvement and to predict potential issues before they occur. For mRNA, digital twins could be deployed for processes, such as microbial fermentation to generate pDNA, in vitro transcription to produce mRNA, and formulation.

A True One-Stop-Shop Provider

The key to success for contract development and manufacturing organizations (CDMOs) is to listen carefully to their customers regarding their needs and to then create the optimal solutions that address those pain points. One of the biggest issues for mRNA developers today is the fragmented nature of outsourcing services. Often, developers must work with multiple CDMOs that only support one step of the process — pDNA, mRNA drug substance, or mRNA–lipid nanoparticle (LNP) drug product.

Some larger companies have attempted to address this issue by acquiring smaller CDMOs that specialize in each of these areas. However, these acquired businesses often remain as separate companies in different locations, each with its own team. Material must be shipped from one site to the other and be handled by an entirely different team, which can create the same kind of inefficiencies as tech transfers between different CDMOs.

At BIOVECTRA, all aspects of mRNA–LNP manufacturing exist at one facility under one roof. The entire GMP manufacturing train is located in a single building, and client projects from pDNA production to mRNA–LNP fill/finish are supported by the same team of experts. Internal production of pDNA also simplifies the supply chain and gives us greater control over timelines. We have also overcome the ubiquitous bottleneck associated with limited fill/finish capacity by establishing in-house capabilities.

Emphasis on Effective Process Development

Establishing robust processes requires a comprehensive approach to process development. BIOVECTRA has always placed an emphasis on not only creating strong teams but supporting them with great manufacturing science and technology to enable the seamless advancement of programs from process development to GMP manufacturing. This approach positions us to evaluate and implement comprehensive process solutions, including new automation technologies that accelerate process optimization and reduce risks associated with commercial production.

As BIOVECTRA moves to automated approaches, we will continue to rely on a QbD approach. Design-of-experiment (DoE) principles will continue to guide process development efforts, with knowledge about key process parameters that impact impurity generation and product yield used as key inputs into AI algorithms that perform predictive analysis.

Optimizing Scale

BIOVECTRA has constructed a brand-new facility that will come on stream for GMP manufacturing in 2023. We have scales from shake flasks and smaller lab-scale equipment up to the 1,000-liter scale for pDNA and the 30-liter scale for mRNA drug substance and complementary scales for mRNA–LNP drug product. Two separate fill/finish lines provide capacity for over 20,000 vials per day each, which equates to nearly 70 million doses per year of a typical mRNA vaccine.

The offering of sub-liter to 30-liter scales for mRNA drug substance and complementary scales for mRNA–LNP drug product manufacturing was selected on the basis of expected demand for these materials and the nature of the processes involved. Developers of mRNA products are driving extremely hard to improve translational efficiency and reduce dose sizes (e.g., through targeted delivery) to reduce the cost of these drugs and vaccines. In vitro transcription reactions, meanwhile, are typically completed in just three hours, allowing numerous runs to be completed per day. In addition, this type of smaller batch-based approach reduces risk, given the expense of the raw materials.

If there is a need for production of mRNA and mRNA–LNP products in larger volumes, BIOVECTRA is well-positioned to respond. We perform fermentation reactions at up to the 17,000-liter scale — the largest capacity in North America. Indeed, we have extensive experience overcoming scalability challenges for both biologics and small molecule manufacturing, experience that will translate well to mRNA and mRNA–LNP production.

Monitoring Advances in Delivery Technologies

While mRNA technology has been under development for several decades, there are still many opportunities for improvement. As a CDMO offering end-to-end mRNA therapeutic and vaccine services, it is essential that BIOVECTRA maintain awareness of evolving technologies in the field and be prepared to implement new production processes leveraging novel technologies.

One of the biggest areas of innovation today revolves around targeted delivery of mRNA to specific cells and/or tissues. The goal is to find solutions that will eliminate safety concerns around potential negative responses to PEGylated lipids and other components in LNPs and increase the efficiency of delivery, thus reducing the required dosage level.

Potential solutions closely followed at BIOVECTRA include the linkage of antibodies to mRNA–LNPs that can bind to specific receptors on the surfaces of specific cells (similar to antibody–drug conjugates) and the use of alternative forms of RNA, such as circular RNA, that can only be expressed in certain cells or tissue. Other efforts focus on increasing the ability of LNPs to stabilize mRNA, thus prolonging the time the active is present in the body.

With decades of experience in producing many types of biologic and small molecule drug substances and drug products, BIOVECTRA is well equipped to manufacture all different types of RNA modalities formulated in different delivery systems. We can express a large spectrum of biomolecules, perform cross-linking reactions with small molecules, and synthesize a wide range of lipids.

Tackling Analytical Challenges to Ensure Quality

The complexity of mRNA processes contributes to the manufacturing risks, which in part relate to challenges with assaying a wide variety of impurities in addition to fully characterizing the mRNA molecules themselves. Current regulatory guidances emphasize ensuring consistent, high quality of mRNA products, which can be assessed using several techniques, including high-performance liquid chromatography, mass spectrometry, gel electrophoresis, and sequencing techniques, among others. In addition to thorough characterization of the mRNA active, detailed impurity profiling, sterility analysis, and stability determination must be completed.

The in vitro transcription reaction followed by various enzymatic reactions and/or capping reactions creates the need to understand the capping, poly-A tail, and other aspects of mRNA drug substances, as well as the various product-and process-related impurities that can be generated. Many of the methods required are unique to mRNA.

BIOVECTRA’s quality analytics team has developed over 50 in-house methods for determining the structure and quality of pDNA, mRNA, and mRNA–LNP products. These assays are platform driven to the extent possible, with detailed protocols in place to complete any aspects that must be customized for a specific mRNA candidate. Importantly, this work is performed within a robust and proven quality system that has a successful track record with regard to regulatory inspections, including by the U.S. FDA, Health Canada, and Japan’s PMDA.

Staying Abreast of Evolving Regulatory Guidance

The most up-to-date mRNA guidance comes from the U.S. FDA, with the latest update occurring in March 2023. Further updates from the FDA are anticipated in the near future. In addition, the WHO — as well as potentially other national regulatory agencies — is expected to issue new mRNA-specific guidelines, possibly before the end of 2023. The U.S. Pharmacopeia has published a second edition of its guidelines for mRNA analytical methods.

At BIOVECTRA, we recognize that mRNA is a new modality with limited real-world data available. Consequently, it is essential during clinical trials of new mRNA candidates to let the data speak for themselves. As more data are generated, agencies will continue to update guidances and share that knowledge.

BIOVECTRA has an internal regulatory group that stays on top of the latest trends and supports clients in their filing processes. They also understand that our customers are at the forefront of the industry, interfacing with the regulatory agencies and consistently reoptimizing their products based on real-time outcomes of animal studies and clinical trials. By partnering with our clients and gathering their feedback, BIOVECTRA is able to optimize and adjust our processes. In fact, interacting with many different clients gives us many different perspectives and allows us to stay on top of potency, safety aspects, and other regulatory concerns, ensuring that we pursue all activities in a compliant manner.

Research- and GMP-Grade Products

The risks associated with moving from preclinical to clinical to commercial stages for any drug candidate can be magnified if raw materials are not taken into consideration from the outset. One way to reduce that risk is to use phase-appropriate raw materials that are produced using the same processes regardless of whether they are research-grade or GMP-compliant.

To that end, BIOVECTRA offers preclinical materials suitable for animal and toxicology studies and GMP-grade materials for use in clinical trials and commercial production. The only difference between these materials is the additional quality documentation, such as the use of validated analytical methods and validated processes. It should be noted, however, that BIOVECTRA does not offer high-throughput screening services for identification of optimal mRNA sequences. We largely produce GMP material for clients that have selected their lead candidate and are looking for the ideal partner to help them move rapidly move beyond the preclinical stage.

Strategic Approach to Personalized Medicine Manufacturing

The success of the COVID-19 mRNA vaccines has drawn much attention to their potential for the prevention of a wide variety of infectious diseases. Many companies are also focused on the development of mRNA-based cancer vaccines, which by their nature are personalized medicines. Such autologous treatments are much more complex in terms of batch records, traceability, quality control, and other manufacturing aspects compared with traditional vaccines.

BIOVECTRA recognizes the challenges associated with advancing such personalized mRNA products. Automation will be essential to minimizing the timeline from patient biopsy to treatment while ensuring traceability and documentation. BIOVECTRA anticipates implementing digitalization and electronic batch records in the very near future. We are also moving toward digitalization and barcoding of samples to greatly increase both traceability and safety. We are looking forward to supporting clients in that personalized medicine space through these improvements.

Originally published on PharmasAlmanac.com on July 31, 2023

With mRNA therapeutics and vaccines receiving high-profile success, the development space for mRNA products is undergoing rapid and unprecedented growth. However, this sudden surge faces  headwinds owing to the scarcity of contract development and manufacturing organizations (CDMOs) specializing in mRNA production. Even among CDMOs offering mRNA manufacturing services, few providers possess the necessary expertise, capabilities, and scalable platform processes to rapidly manufacture mRNA at high and consistent levels of quality across phases and manufacturing scales. Program success may depend on the application of a concerted quality-by-design (QbD) approach, ensuring that the mRNA is optimized for consistency from small-scale inception and research to large-scale, high-quality GMP manufacturing. Quality and efficiency are established across scales and phases by partnering with a true end-to-end provider able to support all of a program’s needs, from optimization of the RNA sequence to GMP production.

Vernal Biosciences’ Concerted Approach

Taking a concerted quality-by-design (QbD) approach to the development and manufacturing of RNA means producing research-use-only (RUO) materials while keeping the end goal of clinical-grade production in mind.  This can be achieved by adopting manufacturing operations that will ultimately be compatible with large-scale production. Along with this QbD approach and the expertise of veterans in the sector, one of Vernal Biosciences’ strengths is the ability to produce high-purity mRNA and lipid nanoparticles (LNPs) with the shortest turnaround time in the industry partnered with established platform technologies and robust processes. Additionally, the quality metrics established for small-scale production are maintained through large-scale production, ensuring proper product quality approval and consistency from drug discovery research all the way through clinical and commercial manufacturing.

Phase-Appropriate Product Grades

At Vernal Biosciences, our unique QbD approach is reflected in the different project types and product grades produced by the company. Through our custom RUO products and services, Vernal provides turnkey solutions to support clients’ early research and discovery needs for mRNA and LNP testing in cell and animal studies.

For clients moving beyond early-stage R&D and RUO materials, Vernal Biosciences offers mRNA and LNP–mRNA made under GMP-Principle and GMP. Products made under the GMP-Principle are typically suitable for IND-enabling GLP tox studies, are produced using processes that mirror those used for RUO products, and are directly representative of the processes scaled for GMP products, even though not typically made in a GMP clean room. On the other hand, full GMP products are intended for first-in-human use and are produced in full compliance with GMP requirements and accompanied by comprehensive documentation and a GMP data package. GMP and GMP-Principle products are produced under oversight of manufacturing quality that is based on phase-appropriate quality systems and risk-management approaches.

A similar approach is applied with respect to the generation of LNP-formulated RNA products. At the RUO level, Vernal currently generates RNA drug substances encapsulated in LNPs for use in toxicology and other preclinical studies. Notably, RUO-grade mRNA can be generated through in vitro transcription (IVT) at a relatively large scale in terms of laboratory quantities: as much as 500 mg.  Vernal anticipates offering GMP mRNA and LNP formulation services in 2023 and expects to further expand operations with a commercial-ready sterile fill/finish offering at a second CDMO site in 2024.

Platform Process with Baseline Quality Targets

Vernal Biosciences’ platform processes for mRNA and LNP–mRNA manufacturing leverage a consistent set of processes and reagents that enable plug-and-play, turnkey solutions. Customers provide the sequence information and the scale needed, while Vernal rapidly and reliably produces RUO materials. Concurrently, RUO design will be generated with efficient scale-up to GMP processes that mirror those used for RUO. This end-to-end process design eliminates many of the tech transfer and proof-of-concept challenges faced when changing CDMO partners as the life cycle of drug development advances. Confidence in our platform processes and manufacturing expertise is based on our experience in manufacturing hundreds of batches of RNA and LNP formulations.

The establishment of platform processes at the RUO stage begins with codon sequence optimization to enhance protein expression. This process factors in accumulated knowledge, evidence, and studies on the effects of different DNA template elements on IVT and translation efficiency. This enables the establishment of critical quality attributes (CQAs), such as the characteristics of the pilot RNA batches generated via IVT and the type and level of certain product-related impurities, which can be determined using capillary electrophoresis (CE). After the subsequent capping reaction (for which Vernal Biosciences uses enzymes rather than other proprietary capping agents) and purification, CE is repeated to confirm the purity of the final capped mRNA product. For special projects that involve other types of RNA, such as structured or circular RNA, additional CQAs will be established and analytical methods will be developed, tested, and performed for every production run.  

Determination of CQAs at the research-grade product level establishes a baseline target for product quality and RNA integrity. Once that is achieved at the drug discovery stage or for RUO materials, a quality target is established, which becomes the initial goal during scale-up. Additional CQAs are layered on to support the quality target product profile (QTPP) for each specific product. 

Process Development and Optimization Begin at the Research Stage

At the RUO stage, it is possible to use the same or similar platform processes for different RNA projects, as these products are typically produced at small scale for RUO. However, differences in RNA sequences, including length, GC (guanine–cytosine) content, and other factors, can have significant impacts on process efficiency and ultimately on manufacturing costs and timelines. As a result, process development and optimization become critical to adapting the platform processes used for RUO into efficient, scaled-up processes suitable for GMP-Principle and GMP products.

Process optimization should be a focus as early in the project timeline as possible. At Vernal Biosciences, the first step is to screen the RNA sequence to verify that it does not contain motifs that might present issues. However, understanding about how different constructs will behave during IVT is still evolving. When odd behavior is observed, Vernal develops hypotheses regarding the cause of the behavior, such as the presence of repetitive sequences or DNA–RNA associations.

Once a hypothesis has been established, a series of small-scale trial runs is performed to determine which modifications improve the productivity or product-related purity profile, which can enrich our existing knowledge base and enable better prediction of the construct’s behavior for future projects. Additionally, if a product must be shipped from one facility to another, the logistics of storage, as well as the freezing and thawing process, must also undergo evaluation.     

Vernal’s platform processes are designed from the start both to require little to no modifications to produce RUO materials and to transition seamlessly, with some modifications, into GMP manufacturing. In most cases, the types of adaptations required as a process advances to GMP can be anticipated on the basis of the information gathered during the RUO phase, such as transitioning to a larger bioreactor, extending the reaction time to increase productivity and yield, and shortening the reaction time to avoid overwhelming the chromatography column. 

For many CDMOs, projects are brought in at the later development stages, with feasibility work completed and important information generated elsewhere. As a result, processes are not necessarily designed to fit the CDMO’s production platforms (if platforms are available), leading to the need for extensive adaptation through an elaborate technology transfer process. At Vernal Biosciences, our end-to-end capabilities span early- and late-stage development, as well as activities from production of working cell bank, plasmid DNA, and RNA drug substance to LNP formulation and filling into vials, which allows a high level of communication between the research and GMP operations groups.

Scaling Considered from the Outset

Despite extensive optimization efforts, some changes in process performance can be expected during scale-up. When developing RNA drug substance and final product (LNP–mRNA) manufacturing processes, it is crucial to know which parameters will be impacted by scale and to plan accordingly. 

Scale-up can be achieved in two ways: increasing bioreactor volumes (using a bigger reactor and more raw materials) or improving productivity. A more productive IVT reaction yields a higher quantity of RNA material per unit volume. Increasing productivity is a focus for Vernal’s RNA Science team, while increasing scale by moving to bigger bioreactors is an emphasis for later-stage products. Once a process at a very small scale has been optimized with respect to productivity and has been demonstrated to work with most constructs, it is ready to be scaled up in stages from 1–2 mg up to 50 mg and larger.

Vernal is developing and qualifying a toolbox of testing methods. This includes assays that are not applied at the research level, because the relevant impurities are known to change at large scale and thus are only tested upon the initiation of process development, engineering, and GMP runs. They also include methods for the detection of impurities for special projects, such as tests for the presence of double-stranded RNA.

Meeting the Needs of mRNA Developers

The concerted QbD approach that drives Vernal Biosciences’ operations meets the needs of therapeutic developers for reliable and high-quality RNA supply. With our end-to-end service offering from cell banking and plasmid manufacturing capabilities to mRNA and LNP formulation, customers only need to partner with a single CDMO across the entire mRNA value chain for the life cycle of drug development. Vernal’s reliable platform processes, state-of-the-art equipment, and expertise in process development enable seamless scale-up, fast tracking your drug development by ensuring timely and stable supply of high-quality mRNA-based drug products. Vernal is one of the few CDMOs with manufacturing slots available for new projects, as demand for CDMO services in the mRNA technology sector surges to unprecedented levels and mRNA developers struggle to find manufacturing partners with sufficient available capacity.

Originally published on PharmasAlmanac.com on January 10, 2023