Multimodal Biomanufacturing: What It Is, What It Isn’t, and the Attributes that Drive Facility Design

Multimodal manufacturing is a strategic approach that harnesses the power of diverse manufacturing methods to optimize and streamline the productionprocess, specifically within the realm ofhuman therapeuticsin the Advanced Therapy Medicinal Products (ATMP)space. This concept involves integrating different manufacturing techniques/platformsto improve efficiency, reduce costs, and enhance the overall quality of the manufacturing processand the eventual therapeutic product slated for licensure.In the recently issued White House report, “Bold Goals for US Biotechnology and Biomanufacturing” ia prominantobjective isto test and de-risk new biomanufacturing practices for next generation biotechnology products in commercial manufacturing facilities.iiThe growing focus in cell-based therapies, a key component of the next generation of human therapeutics, has led many organizations to focustheir development efforts into amultitude of platforms.Multimodal is not new. For the ATMP space, it was identified back in 2017 as a viable manufacturing option for clinical needs. In the context of the biotechnology ATMP space, multimodal manufacturing techniques are being applied to optimize the production of complex biological compounds and therapies that are studied in multi-omic research, such as gene editing.iiiInreality,many contract development and manufacturing organization (CDMOs)have been operatingin a quasi-multimodal manner for decades. Multimodal can include vaccines, monoclonal antibodies, viral vectors, plasmids, allogeneic cell therapies, and autologous cell therapies. The challenge is crafting amanufacturing plan that supports your business without attempting to become a one-size-fits-all solution for every need.Market DriversThe forecasted growth of the Cell and Gene Therapy (C&GT) market is significant. The ATMP space, where C&GT continues to serve as the primary catalyst for growth, is predicted to grow at a compound annual growth rate of 39.4% for the period now until 2030.iv(Figure 1)

2Figure 1In this growth projection, cell therapies account for 15% of the growth and gene therapies account for another 30% of growth, with many of these products advancing out of early-stage clinical manufacturing towards commercialization.Just as the 1990s large molecule market saw a “fork-in-the-road” decisionregardingsingle vs. multi-product manufacturing and the technologies to implementv, the ATMP space is grappling with critical questions around asset size, cost, technology implementation, flexibility, scale, and many others. Transitioningto a multimodal manufacturing strategycould very well emerge as the most prudentbusinessand operational solution.Another key driver fueling the multimodal manufacturing discourseis the rapid pace of advancement in research,developing different modes of therapy to address multiple human therapeutic targets. The use of combination therapies using multiple therapeutic modalities has elevated anti-cancer activity while lowering doses of agents, thus reducing side effects.viAs previously referenced, gene editing tools such as CRISPR-Cas, TALENs (Transcription Activator-Like Effector Nucleases), and Base Editinghave brought about multiple therapeutic solutions that are being provided for chronic diseases and oncolytic cancer treatments, as well as advanced CAR-Tand TCR therapies.The results from these development activities could pave the way toa number of different manufacturing platforms, including:Viral VectorsCell TherapiesmRNA

3Plasmid Production as starting materialsLNP FormulationOligonucleotidesLooking back again at the history of Monoclonal Antibody (Mab)production, one of the key challenges was the delicate balance of securing themanufacturing capacity when required, whilebeing mindful of the cost of the manufacturing enterprise asset and the time needed to bring it online. There was no universalsolution to manufacturing multiple products in the same facility. Variability in technologies, equipment, scale, and facility attributes had to be addressed. These are the same challenges we face today.Multimodal Manufacturing ChallengesThe foundation elements of any biomanufacturing enterprise are consistent; the Process, the Facility, and the Infrastructure, as defined in figure 2. Each of these elements must focus onarisk-based approach to design that aligns the attributes of the product, the process, and the facility.viiFigure 2

4For different modalities, there will be a number of facility design challenges that will be driven by these attributes. Along with these design challenges, there will also be pressure for the Design Team to address flexibility needs as the product pipeline may shift due to future product manufacturing needs. Segregation StrategySegregation strategy is driven by product protection. Presently,product protection focuses on establishing methods that yielda closed, validatable process that mitigates product contamination risk. But the reality today is that not all process unit operations are closed, so the traditional approaches to segregation strategy still apply.Physical segregationEnvironmental segregationTemporal segregationProcedural segregationVirtual segregationLooking at physical segregation, the modality will impact the attributes of the design. Plasmids are used in many C&GT applicationsas both starting materials and some intermediates. The pDNAis readily isolated from bacterial cells, engineered to express the protein(s) of interest in a mammalian host cell. However, microbial fermentation operations, used in production of the plasmid,introduce a potential manufacturing contamination risk. The facility design must addressthesehigher contamination risks, necessitating, a higher biosafety level and level of segregation would be introduced, including unidirectional flowsandsegregated manufacturing areas.Viral Vectorsare highly leveraged for the delivery of “genetic payloads” in C&GT manufacturing. A viral vector is essentially a stripped-down virus that retains the ability to enter into cells and deliver the genetic payload. The introduction of a virus poses enhanced containment requirements, often mirroring theviral clearance activities for MAbs. The facility design must account for thisincreased contamination risk.Lipid Nano Particles (LNP)are utilized in mRNAdelivery, where they stabilize and protect the mRNA. Giventheir synergy, stability, and attribute alignment, the segregation strategy of the facility design would be the samein terms segregation and flows.Oligonucleotidesare a class of ATMPsthat are used in gene and protein expression.There a number of manufacturing challenges around Oligo manufacturing. One is the use of solvents in the manufacturing process, which results in a higher level ofhazardous process waste that must also be addressed. Given the safety concerns associated withsolvents, the segregation strategy must provide a solution for managingflammables and solvents.

5HVACDesign/Area ClassificationFor a growing number of ATMP facilities in the C&GT space, a key challenge is pinpointingrisk mitigation strategies that target contamination control. A direct outcome of these efforts is the potential reduction of area classifications within the manufacturing spaces,consequentlysimplifying the segregation strategy and optimizing the overall HVAC system design.The commonly used HVAC configurations for ATMP products arethreebasic alternatives applicable to Cell Therapy, Vector Manufacturing and related, cell culture-based products. These alternatives are: Primary/Secondary using AHU/FCU based local recirculation Primary/Secondary using FFU based local recirculationPrimary only, with no recirculationIn a detailed design analysis on the applicability of HVAC system design for ATMPs in Grade B spaces, the study supported the use ofrecirculated HVAC systems. These systemsweredeemedappropriate and capable of meetingthe defined global regulatory expectations that have been traditionally tied to once-through air systems.viiiBy minimizing the HVAC design criteria and reducing area classifications through system design optimization, each modality can be “grouped” under a simplified classification approach:Grade B –Cell TherapyGrade C –Gene Therapy, LNPGrade D –Plasmids, mRNA, OligonucleotidesOperational IntegrityWith different modalities, there will be operational issues that should be reviewed and addressed. Theseinclude:Uni-directional or bidirectional flows to support operational and segregation strategyManufacturing OperationsHazard AnalysisWaste handlingContainment strategy for BiosafetyFor example, Lipid Nano Particle (LNP)production involves water-miscible solvents such asethanol, methanol, or acetone. The use of these solvents requires special considerations for storage and handling, dependent on the volumes being used in the process. Current building/safety codesmandate that facility design attributes takethese requirements into consideration.Process Closureand the Validation ApproachA key success attribute in multimodal facility design must focus on a validated, closed process as previously mentioned. It is essential to tailor the validation approach to the specificproduct and

6process in question. Validation requirements may vary based on the complexity and criticality of the product, the potential risks involved, and applicable regulatory guidelines.To ensure a comprehensive,robustvalidated approach, it’s imperative to consider the following key points:1.Risk Assessment: Performa comprehensive risk assessment to identify potential process closure failure modes. This involves assessing the process steps, equipment, materials, and personnel involved in the manufacturing processunit operations. The goal is to identify any potential risks that could compromise product quality, safety, or efficacy.2.Failure Mode and Effects Analysis (FMEA): Conduct an FMEA to prioritize and evaluate the identified failure modes based on their severity, occurrence, and detectability. This analysis helps in understanding the most critical failure modes within the facility that would require attention.3.Design and Control Strategies: Develop appropriate design and control strategies to mitigate the identified risks. This may involve modifying equipmentorprocess parameters,as well as potentiallyimplementing additional controls and monitoring systemsbased on the process attributes.4.Process Validation: Conduct process validation studies todemonstrate that the implemented design and control strategies are effective in preventing or minimizing process closure failure modes. Process validation typically includes these three stages: process design, process qualification, and continued process verification.5.Process Monitoring and Control: Implement robust monitoring and control systems to continuously monitor critical process parameters(CPP)and performance indicators. This would include the use of process analytical technologies (PAT) and real-time process monitoring to promptly detect any deviations.6.Quality Management System(QMS): Establish a comprehensive QMSto ensure adherence to standard operating procedures (SOPs), good manufacturing practices (GMPs), and other relevant quality standards.7.Training and Personnel Qualification: Ensure that all personnel involved in the manufacturing process are adequately trained and qualified to perform their tasks effectively and in compliance with the required quality standardsfor each defined modality.8.Change Control: Implement a rigorous change control system to assess the impact of any process changes on process closure failure modes. Changes to the process should undergo thorough evaluation and validation before implementation.9.Regulatory Compliance: Ensure that the validation activities and documentation comply with relevant regulatory requirements, such as those from the Food and Drug Administration (FDA) in the United States or the European Medicines Agency (EMA) in Europe.

7A Multimodal Design ApproachA multimodal manufacturing facility refers to a manufacturing asset that can address different manufacturing processes/platforms as elaborated upon previously. For organizations such as Contract Manufacturing Organizations (CMOs), the decision to establish a multimodal manufacturing facility should be based on a thorough assessment of theirstrategic goals, market dynamics, and their technological competencies.Considerationsunderpinning such a decision include:Speed to Market:Different manufacturing modes can offer varying levels of speedto market for some products. By having a multimodal facility, a manufacturing organization can choose the most suitable manufacturing approach for each productplatform, optimizing productiontimes and reducing time-to-market for new products.Cost Efficiency:Depending on the specific product and production requirements, different manufacturing modes can offer cost advantages. A multimodal facility empowersaCMO to choose the most cost-effective manufacturing method for each product, optimizing operational expenses.Flexibility for Market Changes:Consumer preferences and market trends can change rapidly. A multimodal manufacturing facility equips an organization with the agility to swiftlyadapt to shifting demands and evolving market landscapes.Supply Chain Resilience:Having multiple manufacturing options in-house can enhance supply chain resilienceand minimize potential risks associated with timing, especially for certain autologousCell Therapy (CT)products.If one manufacturing process faces disruptions or supply chain challenges, the company can then pivot to another mode to ensure a consistent supply of products.Innovation and Product Differentiation:A multimodal facility allows for the integration of innovative manufacturing techniques, which can lead to the development of unique and differentiated products. This can give a company a potential competitive edge in the market and attract customers seekinginnovative and novel productsand manufacturing capabilities.This facility design approach must focus on the specific facility attributes previously discussed. To illustrate this, let’s examine a few layout scenarios:Oligonucleotides (Oligo)For an Oligomanufacturing area, akey concern as mentioned is the use of solvents. This points to a layout where key equipment will be placed on outside walls in order to comply with building and safety codes for hazardous materials (class 1, division 1)ixOligo is also more likely to implement bidirectional flow to improve efficiency during manufacturing, and will not require any form of biosafety classification or higher-level cleanliness classification for product protection (Grade D).

8Figure 3:Oligonucleotide LayoutmRNA and LNP

9While these are two distinct components used in the manufacture of mRNA-based therapeutics, they are both critical to the success of providing the therapeutic benefit to the patient. In simple terms, the mRNA provides the genetic instructions for cells to produce a specific protein, while the LNP encapsulates and delivers the mRNA into cells, protecting it from degradation and facilitating its uptake.Neither mRNA or LNP requires special biosafety considerations. Nor do either rely on a need for unidirectional flows or higher-level cleanliness classifications. However,as with Oligo products, LNP manufacturing requires the use of water-miscible solvents in a buffer solution. Figure 4: mRNA and LNP Layout

10SummaryMulti-modal manufacturing is progressively becoming apart of the ATMP manufacturing landscape. In a recent industry survey, more than half of survey respondents plan to implement multimodal solutions within the next two years (Figure 5)x.Figure 5Much of this burgeoning interest stems from the substantial value that can be unlockedby implementing this type of manufacturing approach. For example, a growing number of CMOs are developing plasmid production capabilities to support their mRNA manufacturing needs. Simultaneously, they are establishing themselves asthird-party suppliers of plasmids to external customersfurther diversifying their revenue streams.Several factors are driving momentum:1.Flexibility: As demonstrated, multimodal manufacturing platforms can be designed with flexibility in mind, allowing for the incorporation of new processes or modifications to existing processes as scientific understanding advances.2.Cost Efficiency: Consolidatingmultiple manufacturing steps and technologieswithin a single facilitytranslates into enhancedoverall process efficiency, potentially reducing costs associated with labor, equipment, and materials.3.Integration of Experience: Multimodal manufacturing encourages cross-disciplinary collaboration, enabling expertsfrom different disciplinesto contribute their knowledge to the development of a unified manufacturing operation.

114.Time-to-Market: An argument could be made that multimodal manufacturing can accelerate the production timeline by eliminating bottlenecks and inefficiencies that may arise when using disparate manufacturing processes.In conclusion, multimodal manufacturing is becoming a cornerstone of the ATMP manufacturing landscape, driven by its inherent value and adaptability. As industry leaders increasingly harness its potential, itpromises to propel the ATMP sector toward greater efficiency, innovation, and responsiveness to evolving market demands.iBold Goals for US Biotechnology and Biomanufacturing: Harnessing Research and Development to Further Societal Goals”White House, Executive Order 14081, March 2023.iiIbid, p. 43.iiiIbid, p. 38.ivBioSpace Forecast Report, August 11, 2022. https://www.biospace.com/article/cell-and-gene-therapy-market-size-growth-trends-forecast-report-2022-2030/vWitcher, Carbonell, Bigelow, Lewis, Zivitz, Odum; “Facility of the Future: Next Generation Biomanufacturing Forum” Parts I, II, III, Pharmaceutical Engineering, volume 33, No. 1, 2, 3, 2015.best business/operational solution.viKemp, Shim, Heo, Kwon, “Co-delivery of multimodal therapeutics for efficient, targeted, and safe cancer therapy”, University of California –Irvine. https://www.sciencedirect.com/science/article/pii/S0169409X15002446viiProcess Architecture in Biomanufacturing Facility Design, Wiley and Sons, Odum and Flickinger, 2018, ch. 2viiiGenesis Engineers, “Technical opinion: Applicability of Recirculated Air for ATMP Products in Grade B/ISO 7 and Grade C/ISO 8”, March 2022ixUS Department of Labor, OSHA Standards, 1910.106, subpart H, Flammable LiquidsxBioProcessInternational, “Four Design Factors Shaping Multimodal Cell and Gene Manufacturing,” bioprocessinternational.com, June 18, 2021.