Cell Culture Dish Podcast

As demand for DNA and RNA therapeutics continues to accelerate, manufacturers are under increasing pressure to improve efficiency, reduce risk, and scale production without expanding facility footprints. One emerging solution is the integration of traditionally separate downstream steps into a single automated platform. Asahi Kasei Bioprocess, has addressed this need with the THESYS® platform, a suite of oligonucleotide manufacturing technologies designed to streamline workflows from synthesis through downstream processing. In a recent podcast, Sagar Bhatt, Senior Project Engineer at Asahi Kasei Bioprocess America, discussed the development of an integrated system within the THESYS C&D/TFF system combines cleavage, deprotection (C&D), and tangential flow filtration (TFF), and why this shift represents a meaningful evolution in oligonucleotide manufacturing. Rethinking a Fragmented Workflow Historically, oligonucleotide production has relied on a series of disconnected unit operations. Cleavage, deprotection, ultrafiltration, and diafiltration are often performed across multiple systems, sometimes even in different rooms. “Cleavage and deprotection are often carried out using fairly basic setups… and in many cases, they still involve a lot of manual handling,” Sagar explained. “They also typically require additional equipment, like separate tanks, which adds complexity to the process.” This fragmented approach introduces several challenges. Material transfers between systems increase processing time and create opportunities for product loss. In addition, deprotection reactions, particularly for RNA, require careful thermal control due to heat generated during acid addition. “If the rate of the acid addition and resulting temperature rise are not controlled carefully, it can negatively impact the product… and damage product quality.” Facility constraints add another layer of complexity. Because oligonucleotide processing often involves flammable solvents, operations must occur in hazardous environments. However, traditional filtration systems are not typically designed for these conditions, forcing manufacturers to physically move material between areas. Recognizing these inefficiencies, Sagar and his team saw an opportunity to simplify. “By integrating these operations into a single physical equipment and related automation, we could potentially streamline the workflow, reduce handling steps, and significantly improve overall manufacturing efficiency.” From Concept to THESYS® Integration The idea of combining reaction-based and membrane-based processes might seem complex, but Bhatt emphasized that the separation of these steps is largely historical, not technical. “Cleavage and deprotection are reaction steps… whereas TFF is a membrane separation process,” he said. “Though there are different mechanisms involved, they can operate on the same product stream and can share the same fluid handling architecture if designed properly.” Within the THESYS® platform, this integration is enabled through automation and system design that bring multiple unit operations into a single, cohesive workflow. Advances such as closed-loop temperature control, precise dosing, and real-time pressure monitoring allow both reaction and filtration steps to be managed within one system boundary. Equally important was designing the platform for hazardous environments from the outset. “That eliminated the need for intermediate product transfers, which made this integration approach much more practical.” Engineering for Efficiency and Scale One of the most significant engineering challenges was balancing performance with practicality. “Designing the combined system to keep the footprint to a minimum while also taking operability and maintainability into consideration was one of the biggest challenges,” Sagar said. The team also focused on minimizing holdup volume, maximizing product recovery, and ensuring cleanability for GMP operations—all within a compact system design aligned with THESYS®’s broader focus on efficient, scalable manufacturing systems. The result is a platform that delivers efficiency gains primarily by eliminating transfers. “In traditional setups, the material moves between different systems and sometimes even between different rooms,” he explained. “Each transfer adds time, manual handling, and potential product loss.” By consolidating operations into a single THESYS-based system, manufacturers can complete processes sequentially without interruption, reducing both time and risk. Improvements in Safety and Process Control Beyond efficiency, integration significantly enhances safety and control. “Operators no longer need to move material between systems in solvent handling environments,” Sagar said. “Keeping everything inside one enclosed physical system significantly reduces exposure risk and handling errors.” From a control standpoint, a unified automation framework governs all stages of the process. “The same system carefully controls dosing and temperature during the reaction phase and also controls pressure and flow during filtration,” he noted. “The unified control improves reproducibility and helps contain the process within validated operating conditions.” Footprint and Operational Savings The benefits extend to facility design as well. “Instead of installing separate systems with their own vessels, pumps, and control panels, manufacturers operate one integrated system,” Sagar said. “This can free up valuable clean room space and reduce infrastructure requirements.” Fewer systems also mean fewer cleaning cycles, fewer validations, and less maintenance, resulting in meaningful operational savings over time. Designing for Flexibility: A “Built for You” Approach A defining characteristic of the THESYS® platform is its flexibility. The system is designed to align with each customer’s process rather than enforce a rigid standard. “The system is designed to adapt to each customer’s manufacturing process rather than forcing the customers to adapt their processes to the equipment,” Sagar explained. For example, the system can accommodate different filtration control strategies. “Some customers perform filtration using transmembrane pressure… while others use permeate flow. The system has the capability to operate on both.” Customer feedback also shaped physical design elements, including vessel sizing, filter configurations, and facility integration. “These kinds of small but critical considerations help the system fit naturally into the customer’s manufacturing environment without major disruptions.” Validation, Compliance, and Integration Integration also simplifies GMP validation. “When you combine multiple operations into a single system, you reduce the number of vessels, process flow paths, and connection points,” Bhatt said. “That means fewer product contact surfaces to clean and validate.” The system uses hygienic design principles and recipe-driven automated cleaning cycles, with electronic batch records supporting compliance. Importantly, the THESYS®-based system integrates easily into existing manufacturing lines. “The upstream synthesis process does not change,” Sagar noted. “After synthesis, the product simply enters this integrated platform.” Because the system is designed for hazardous environments, it can be installed directly where cleavage and deprotection already occur, eliminating the need for downstream relocation. Industry Recognition and Future Impact The system’s impact has already been recognized with an Interphex Innovation Award, which Sagar described as “a strong validation from the industry.” “The award recognized that integrating these steps… addresses real challenges that manufacturers face, like complex workflows and inefficient equipment setups.” Looking ahead, he sees integration—central to platforms like THESYS®—becoming a core design principle in oligonucleotide manufacturing. “As demand for DNA and RNA therapeutics continues to grow, manufacturers will need equipment that supports higher throughput, consistent product quality, and faster batch turnaround.” Integrated, modular platforms will play a key role in meeting these demands. “Over time, we are likely to see more compact automated systems performing multiple process steps… helping facilities scale production more efficiently and accelerate the development of new therapies.” A Shift Toward Smarter Manufacturing The integration of cleavage, deprotection, and TFF within platforms like THESYS® represents more than just a technical advancement, it signals a broader shift toward streamlined, automated, and modular biomanufacturing. By reducing complexity, improving safety, and enabling scalability, integrated systems are poised to redefine how oligonucleotides are produced, bringing the industry closer to faster, more efficient delivery of next-generation therapeutics. To learn more, please see Optimize Your Oligo Manufacturing

Surfactants are indispensable in the production of biologics, vaccines, and cell therapies. Yet for years, they’ve posed a persistent challenge: they are notoriously difficult to monitor accurately and in real time. That challenge is now being addressed by Nirrin and its groundbreaking Atlas platform, a real-time spectroscopy solution that is reshaping how biomanufacturers measure and manage surfactants. In this episode of The Cell Culture Dish podcast, Editor Brandy Sargent spoke with Bryan Hassell, Founder and CEO of Nirrin, and Hannah Furrelle, Analytical Scientist at the company, to discuss the science behind Atlas and its implications for bioprocessing. Real-Time Data Without Compromise At the core of Atlas’s innovation is its ability to provide high-quality quantitative data in under a minute—without any sample preparation. “The real breakthrough with Atlas is speed with confidence,” explained Hassell. “Time to market for biopharma is increasingly critical, yet a lot of critical decisions still rely on data from assays that take days or even months. Atlas changes that.” Unlike traditional techniques, which often require significant sample manipulation and suffer from matrix interference, Atlas uses high-precision tunable laser spectroscopy to directly analyze samples in their native form. “What makes Atlas so powerful is that we’re looking at the sample without altering it,” Furrelle explained. “That means the data we get is true to the process—there’s no distortion from prep steps or artifacts introduced by the method.” Moving Beyond PLS: A New Modeling Approach One of the technological breakthroughs enabling this leap in performance is Nirran’s move away from PLS models in favor of an iterative optimization framework. This approach eliminates the need for extensive training data, reducing model complexity while increasing robustness and flexibility. “Where a PLS model might need 20 to 30 bioreactor runs to build a dataset, Atlas delivers data on the fly,” Hassell said. “It’s not only faster, it’s more robust, more compliant, and more versatile—especially for applications like scale-up or tech transfer, where traditional models often break down.” Applications Across the Biomanufacturing Workflow Atlas is already being integrated into real-world bioprocessing environments, including both batch and continuous manufacturing. In batch processes, manufacturers use Atlas to confirm critical parameters—like protein and excipient concentrations—before proceeding to the next unit operation. This enables earlier course corrections and helps prevent downstream failures. “In the past, you either waited days for lab results or moved forward at risk,” said Hassell. “Atlas provides the immediate answers needed to make confident decisions in the moment.” For continuous manufacturing, the value is equally profound. Atlas provides the real-time, quantitative feedback necessary for dynamic process control. “You can’t have continuous processing without real-time data,” he said. “Atlas gives you the insights needed to support real-time decisions at every step.” Eliminating Risk with No-Prep Analysis One of Atlas’s standout features is its ability to deliver no-prep analysis. This eliminates sources of variability that often arise during sample handling and processing. “We’re scanning samples in their native form,” said Furrelle. “That means what we’re measuring reflects what’s actually in the process—without distortion from dilutions or centrifugation.” This no-prep capability also speeds up workflows and eliminates risk by allowing operators to verify component concentrations instantly before committing to the next step in production. Laying the Foundation for Smart Biomanufacturing Nirrin sees Atlas not just as a data tool, but as a stepping stone to smart biomanufacturing. Although full automation isn’t yet widespread, Atlas is helping to lay the groundwork by delivering trustworthy real-time data, something most operations have historically lacked. “Right now, we’re focused on validating the technology and educating the industry,” said Hassell. “Without sensors like ours, you can’t have smart manufacturing. But once real-time data becomes available, everything else,automation, digital twins, AI,can start to fall into place.” Furrelle agreed, adding, “You can’t automate without sensors. Atlas gives you real-time insights that teams can actually use, not work around.” From QC Tool to Strategic Platform As teams adopt quality-by-design (QbD) approaches, Atlas is being used well beyond its initial QC role. It’s becoming a platform for optimizing surfactant levels, improving batch-to-batch consistency, and proactively preventing formulation issues. “You can’t have quality if you don’t start from a place of quality,” Furrelle said. “Validating components before you use them is no longer just ideal, it’s achievable.” Supporting Regulatory Compliance and GMP Readiness With growing expectations around regulatory compliance, Atlas offers distinct advantages for GMP workflows. By capturing data directly from the process, without modification, it ensures traceability, eliminates method error, and delivers reproducible results. “Getting data that wasn’t previously accessible is foundational to compliance,” said Furrelle. “Most of the time, we assume buffer concentration is correct based on scale readings and signatures. With Atlas, you can actually measure and prove it.” Hassell added that Atlas’s direct-measurement capability enhances confidence in audits and inspections. “When you can take a sample straight from the line and analyze it without modification, you eliminate layers of error. That’s critical for compliance and it’s something our customers are really gravitating toward.” Built for Sustainability and Greener Excipients Atlas also aligns with the industry’s increasing focus on sustainability. Its optical system requires no consumables, a rare feature in today’s single-use-dominated industry. “We deliberately avoided a consumables-based model,” Hassell said. “Our customers want tools that align with their long-term goals and sustainability is a big part of that.” The platform also supports real-time validation of novel excipients, including greener alternatives. Its broad spectral range is packed with chemical information, enabling confident measurement of even chemically distinct surfactants. “We’ve had great success supporting teams developing new excipients,” said Hassell. “The system’s flexibility and depth of information make it future-proof.” Real-World Impact: Surfacing Hidden Issues Early use cases illustrate the power of the platform. In one of the first beta tests, a biopharma team used Atlas to trace a suspected surfactant stability issue. “They didn’t know where the problem started, but within minutes, we helped them pinpoint exactly where the formulation went off track,” said Hassell. “It was a powerful example of real-time debugging.” Furrelle recalled a similarly impactful use case involving ultrafiltration and diafiltration (UF/DF). “Traditionally, you wouldn’t know if you lost or enriched an analyte until after the step was complete. Atlas allows you to monitor the process live and optimize it. That’s a huge advance.” A Broader Role in Process Development With increasing adoption, Atlas is quickly evolving into a comprehensive process platform. “I’ve always thought of it as more than QC,” said Furrelle. “It’s helping teams move from retrospective quality control to proactive process design.” More teams are now asking not just whether a step worked, but how to improve it. Atlas provides the data needed to make those decisions in real time. What’s Next for Nirrin and Atlas As of now, Nirrin is focused on validating the technology and supporting its integration into real-world workflows. With backing from the NIH and a growing customer base, the company has no shortage of momentum. While the current focus is on protein-based therapeutics, Nirrin is already preparing for broader applications. “We have line of sight into other modalities, like cell and gene therapies,” said Hassell. “But first, we want to solve the problems in front of us really well and help the industry move forward with confidence.” Looking Ahead: The Future of Surfactant Analytics Both Hassell and Furrelle are confident that Atlas will play a central role in the future of surfactant monitoring. “We want Atlas to be the go-to instrument for companies developing new formulations,” said Hassell. Furrelle emphasized the importance of robust analytics during tech transfer. “Right now, that hand off is often based on assumptions and broad specs. With Atlas, you can measure exactly what’s happening and that changes everything.” Conclusion Atlas isn’t just solving an old problem, it’s setting a new standard. With its speed, accuracy, and no-prep design, it’s enabling smarter biomanufacturing, better compliance, and more sustainable practices. As the industry moves forward, Atlas stands poised to become an essential platform for quality, innovation, and transformation in bioprocessing. See Atlas in Action: https://www.nirrin.tech/nirrin-atlas-evaluation-contact-us/

In this podcast, we spoke with Dr. Jorge Escobar Ivirico, Product Manager, Bioprocess Solutions at Eppendorf, about the fascinating world of induced pluripotent stem cells (iPSCs), exploring their groundbreaking potential in regenerative medicine, personalized therapies, and drug development. Our guest explained how iPSCs, created by reprogramming adult somatic cells, can differentiate into virtually any cell type, making them invaluable for research and therapeutic applications. We delved into the importance of consistency, quality control, and reproducibility in iPSC production, alongside the challenges of culturing these cells, such as maintaining pluripotency and scaling production for clinical use. The discussion highlighted exciting advancements, including the development of organoids and universal T cells, as well as the ethical considerations distinguishing iPSCs from embryonic stem cells. Looking to the future, Jorge envisioned iPSCs becoming a cornerstone of standard medical practice, while acknowledging the need to address safety, scalability, and regulatory hurdles to fully realize their potential. What are Induced Pluripotent Stem Cells (iPSCs)? "Induced pluripotent stem cells are a type of stem cell created by reprogramming adult somatic cells, like skin or blood cells, back into an embryonic-like state," explains Jorge. This process involves introducing specific transcription factors, often called Yamanaka factors, to transform these cells into a versatile state. Once reprogrammed, iPSCs can differentiate into almost any cell type, making them invaluable tools for research, drug development, and potentially life-changing therapies. The Growing Importance of iPSCs iPSCs offer a range of advantages, particularly their ability to sidestep ethical concerns tied to embryonic stem cell use. “What makes iPSCs so important today,” Jorge notes, “is their versatility and potential applications. Researchers can create patient-specific cell lines, which are essential for drug screening, disease modeling, and personalized medicine.” This technology is pivotal for regenerative medicine, offering hope for repairing damaged tissues and organs. “From neurodegenerative diseases to heart damage, iPSCs open the door to innovative treatment possibilities,” he adds. Mastering the Production Process Producing iPSCs is a meticulous endeavor. "Consistency is key," emphasizes Jorge. Researchers must ensure that each batch of cells meets strict criteria to avoid unpredictable outcomes, especially when precision is vital in both research and therapeutic applications. Standardized protocols and quality control measures are essential to achieve consistency. These involve monitoring for contamination and verifying the cells' ability to differentiate into various cell types. “Imagine developing a therapy based on a specific batch of cells, only to find that subsequent batches behave differently,” he warns. “Such inconsistencies can jeopardize patient outcomes.” Tackling Challenges in Culturing iPSCs Culturing iPSCs presents its own set of challenges. High cell numbers are often needed for large-scale research or therapeutic applications, but scaling up production without compromising quality is no small feat. Maintaining the cells’ pluripotent state is another hurdle, as they can easily differentiate prematurely under certain culture conditions. "Environmental parameters like temperature, pH, oxygen levels, and nutrient availability must be rigorously controlled," Jorge explains. “Even minor fluctuations can negatively impact cell health and their ability to remain pluripotent.” Innovations Addressing Culturing Hurdles To overcome these challenges, researchers are turning to advanced techniques like 3D culture systems and bioreactors. These provide a more natural growth environment for the cells, enhancing their viability and functionality. “By transitioning from traditional 2D cultures to 3D systems, we can better mimic the natural environment, significantly improving outcomes,” Jorge shares. 3D systems also increase growth surfaces for cell attachment and allow for higher cell density, which is critical for producing the large quantities required for therapies. "It’s not just about quantity," he notes. "These systems help maintain the quality of the cells, ensuring consistent results across labs and facilities worldwide." Transformative Applications of iPSCs One of the most exciting applications of iPSCs is in creating universal T cells for therapies like CAR-T cell treatments. “Traditional CAR-T therapies require harvesting a patient’s cells and modifying them—a time-intensive process that isn’t suitable for all patients,” Jorge explains. "With iPSCs, we can create a bank of genetically modified T cells that can be used across multiple patients, reducing risks like graft-versus-host disease and speeding up treatment timelines." Balancing Potential with Risk Despite their promise, iPSCs come with risks. “If iPSCs are not fully differentiated before transplantation, they can form teratomas—tumors containing various cell types,” Jorge cautions. Prolonged culturing can also lead to genetic instability, raising concerns about mutations that could affect therapy outcomes. Additionally, advanced techniques like CRISPR gene editing, while revolutionary, carry risks of off-target effects that could lead to unintended consequences. “Ensuring the safety and efficacy of iPSC-derived therapies requires rigorous preclinical testing and long-term monitoring during clinical trials,” he emphasizes. The Role of Embryonic Stem Cells While iPSCs offer an ethical alternative, Jorge argues that embryonic stem cells still have a role in research. "They provide unique insights into early human development and serve as important controls for studying pluripotency and differentiation," he explains. "Each type of stem cell has its own advantages, and we should leverage them where they are most effective." A Glimpse into the Future Looking ahead, Jorge envisions significant advancements in iPSC-based therapies over the next 5-10 years. “We’re likely to see more approved treatments for conditions like neurodegenerative diseases, heart disease, and certain cancers,” he predicts. Improvements in manufacturing and scaling processes will make these therapies more accessible, potentially transforming them from experimental solutions to standard medical practices. Remaining Challenges Achieving these milestones will require addressing several hurdles. "Safety and standardized production protocols are critical," Jorge stresses. Regulatory frameworks also need to evolve to keep pace with these advancements. “The future of iPSCs is incredibly promising, but we must remain diligent in overcoming the scientific, logistical, and ethical challenges ahead.” The field of iPSCs continues to inspire hope for groundbreaking medical treatments, making this an exciting time in regenerative medicine and stem cell research. “iPSCs are truly a testament to human ingenuity,” Jorge concludes.

In this podcast, we spoke with Dr. George Wang, Vice President of Discovery and Preclinical Services at WuXi Biologics about the importance of identifying potential manufacturing, stability, and scalability challenges early to mitigate risks, reduce costs, and streamline drug development timelines. By evaluating factors such as solubility, stability, and manufacturability during initial candidate screening, companies can avoid costly setbacks later in the process. Advanced tools like high-throughput assays, computational modeling, and AI-based predictions are now essential for these evaluations. What Is Developability? Dr. Wang began by defining developability as the assessment of whether a drug candidate possesses the necessary attributes to be scaled up for production during Chemistry, Manufacturing, and Controls (CMC) development and, ultimately, for clinical trials and commercialization. He explained, “It’s about identifying potential red flags early on—issues like aggregation, degradation, or manufacturing inefficiencies—that could derail a candidate further down the line.” Why Focus on Developability During Discovery? Traditionally, discovery efforts have focused on identifying antibodies with the highest efficacy and safety profiles. However, the increasing complexity of biologics, including bispecific antibodies and antibody-drug conjugates, has shifted industry focus. Dr. Wang emphasized the costly consequences of overlooking developability in the discovery phase. “Imagine investing millions into a molecule, only to discover insurmountable stability or manufacturability issues during development,” he said. “Performing these assessments early is like an insurance policy, mitigating risks and saving time and resources.” The Economic Case for Early Developability Assessments Dr. Wang highlighted the economic rationale for incorporating developability assessments during the initial discovery phase. “The cost of discovery is less than 1% of the total development cost. Spending a bit more upfront can save millions in reengineering or restarting development,” he noted. He also pointed out that superior developability attributes can provide a competitive edge, enabling faster clinical trial entry or product approval. Key Challenges and Industry Solutions Despite its benefits, the integration of developability assessments in discovery labs faces challenges. Labs often lack the tools, materials, and expertise required for systematic evaluations. “Developability attributes must be assessed using a robust combination of computational methods, analytical tools, and high-throughput assays, which many labs are not equipped to handle,” Dr. Wang explained. Companies like WuXi Biologics have stepped in to bridge this gap. “Our Discovery unit collaborates closely with our CMC team to identify and address developability issues early on,” said Dr. Wang. WuXi’s “WuXiDEEP™,” platform has become a cornerstone of their success, helping fix more than 50 problematic molecules and guiding hundreds of projects through the development pipeline. A Stepwise Approach to Developability Dr. Wang outlined a stepwise approach to developability assessments, starting with high-throughput evaluations during the initial screening of hundreds of candidates. “We use computational analysis to identify red flags such as post-translational modification hotspots or aggregation risks,” he explained. Promising candidates then undergo more detailed assessments, requiring larger material quantities and lower-throughput methods. Even when issues arise, solutions like protein engineering can salvage candidates with strong biological functions. “It’s not about discarding problem molecules outright but addressing and optimizing their developability profiles,” Dr. Wang emphasized. The Role of AI in Developability Assessments Artificial intelligence (AI) is playing an increasingly significant role in drug discovery, and Dr. Wang underscored its potential in developability assessments. “AI-based modeling and prediction tools enable us to anticipate developability challenges early, optimizing the process even further,” he said. Practical Impact and Case Studies Dr. Wang discussed the webinar he gave, Developability Assessment and Optimization in Antibody Drug Discovery, in which he shared case studies that demonstrated the transformative power of early developability assessments. From rapid decision-making to optimized engineering solutions, these examples illustrated how WuXi Biologics helps clients navigate the complex landscape of biologics development. Final Thoughts As the biologics industry continues to evolve, developability assessments have become indispensable for reducing costs, saving time, and maintaining competitive advantages. Dr. Wang’s insights underscore the importance of integrating this approach during the discovery phase, laying a strong foundation for successful drug development. To learn more, please listen to the full podcast and watch Dr. Wang’s webinar, which provides a detailed guide to implementing developability assessments. Developability Assessment and Optimization in Antibody Drug Discovery

In this podcast, we spoke with Ryan Bernhardt, CEO of Biosero and Jesse Mulcahy, Director and Head of Automation at Cellino about the importance of utilizing automation in cell therapy research and production and the potential of these technologies to transform the healthcare landscape and improve patient access. The Challenge of Accessibility in Cellular Therapy The traditional methods of creating induced pluripotent stem cells (iPSCs) are notoriously laborious and expensive, often costing hundreds of thousands, if not millions, of dollars per patient. This high cost poses a substantial barrier to accessibility for many patients in need of personalized cell therapy treatments. Cellino is leveraging advanced automation, AI, and linear technology to dramatically redefine and improve on traditional production processes. Advancing Automation in Cell Therapy Cellino’s approach employs its innovative technology, known as NEBULA. This system utilizes self-contained units, referred to as cassettes, to cultivate personalized cell therapies directly in hospitals. NEBULA uses AI to monitor cell growth while incorporating laser technology to selectively eliminate unhealthy cells. This level of automation has the potential to reduce the manufacturing costs of personalized stem cell therapies by at least tenfold, making treatments more accessible to a broader range of patients. Supporting automation for Cellino is Biosero’s Green Button Go software suite, which plays a crucial role in automating the workflows of life science organizations. Ryan explained how their technology empowers life science organizations to automate essential scientific processes, facilitating the scheduling of workflows and direct communication with lab instruments. With the capability to run processes continuously—day or night—labs can maintain and cultivate cells without the constraints of a conventional workweek. This 24/7 operational capacity allows for the rigorous demands of cellular therapeutics to be met more efficiently. Bridging Gaps with Integrated Automation Ryan describes how lab automation can no longer be seen as merely robotic arms and conveyor belts; it integrates three key elements: physical, logical, and data. By orchestrating these components, automation streamlines and accelerates research across labs that were traditionally siloed and specialized in specific areas. This approach connects different labs, unifying knowledge, expertise, and data systems, enabling real-time decision-making and data-driven insights. Automation enhances workflows by eliminating delays and optimizing project timelines. It serves as a performance tool for scientists, improving efficiency, consistency, and the ability to address complex challenges, while also incorporating AI and machine learning for smarter, continuous processes. Jesse Mulcahy, Director and Head of Automation at Cellino emphasized the significance of Biosero’s orchestration software in improving efficiency by optimizing scheduling, reducing downtime, and maximizing throughput in cell therapy production. The Green Button Go orchestrator improves consistency by automating key steps and minimizing human intervention, ensuring reproducible results for quality control. The software is flexible and modular, allowing for easy adaptation of workflows as needs evolve, whether adding new instruments or changing protocols. This scalability is crucial for producing personalized cell therapies more efficiently and at a larger scale. Addressing Pain Points and Future Trends Despite the advancements, there are still hurdles to overcome in the biologics’ development landscape. Ryan notes that the field is evolving rapidly, with significant advancements in cell culturing, automation, and decision-making processes. Traditional cell culturing is being automated to assess key factors like cell viability, confluence, and other qualitative aspects, aiding decisions on feeding, splitting, and harvesting. While improvements are still needed, especially for various cell lines and techniques, the integration of cryopreservation and storage/retrieval is also advancing. Newer technologies like spheroids, organoids, and gene/protein therapies are driving automation efforts, with a focus on achieving robustness, cost-effectiveness, and higher quality outputs in life science organizations. Looking ahead, both Ryan and Jesse envision a future of enhanced modularity and integration of AI in biologics development. As more laboratories adopt decentralized manufacturing systems, the potential for on-site production of personalized therapies will drastically reduce costs and time associated with transporting and scaling these treatments. The integration of advanced imaging and robotics technologies will also allow for unprecedented precision in monitoring and manipulating cells, opening new avenues in cell and gene therapy. A Bright Future for Automation in Biologics The future of automation in biologics development appears promising, with the potential to transform the industry and enhance patient access to innovative therapies. Ryan’s parting words capture the excitement of this new era: “It’s hard to think about science being done in the absence of automation.” Meanwhile, Jesse advises organizations to approach automation strategically, starting small and focusing on areas that promise immediate benefits while ensuring team involvement to facilitate smooth transitions. As these technologies continue to evolve, the collaboration between companies like Cellino and Biosero will be pivotal in shaping the future of healthcare, making groundbreaking treatments more accessible to patients in need. To learn more, please see: Cellino's website at https://www.cellinobio.com/ Biosero's website at https://biosero.com/

In this podcast, we spoke with Luca Alberici, Senior Vice President and General Manager, Milan Facility, AGC Biologics about the road to their recent EC and FDA approval to commercially manufacture Lenmeldy™ and their future plans in cell and gene therapy. What is Lenmeldy? We began the podcast by talking about AGC Biologics’ Milan site and their FDA approval to commercially manufacture Orchard Therapeutics’ Lenmeldy. Luca explained that Lenmeldy is a gene modified cell therapy product for the treatment of Metachromatic leukodystrophy (MLD), an ultra-rare hereditary disease characterized by accumulation of fats that causes neurodegenerative symptoms. It is a pediatric disease, and patients generally die by the age of five. With this therapy the patient’s stem cells are collected and modified through the use of a lentiviral vector to add a gene called ARSA that encodes for the right form of the enzyme that these patients are encoding wrong. These modified stem cells are then administered back to the patient so they can immune reconstitute not only the immune system, but the cells will also cross correct through secretion of the right form of the enzyme. After just a single shot of the therapy, there is an improvement in their condition and they develop normally, especially if treated in a pre-symptomatic phase. This is the power of gene therapy at its best. The Pathway to FDA Approval for Commercial Manufacture We then discussed the pathway for receiving approval to commercially manufacture this product and how the AGC Biologics Milan team navigated this process. Luca described that it was quite a long journey. AGC Biologics were manufacturing this product at preclinical and clinical phases dating back roughly 15 years. They worked with a series of different sponsors, it was developed by the San Rafaelle Telethon Institute for Gene Therapy in Milan Italy, then GSK continued the clinical development before it was acquired by Orchard Therapeutics. AGC Biologics remained the manufacturer during this time and in 2020 received approval for commercial manufacture of the product in Europe, but FDA requirements are different so over the last two years, they partnered with Orchard Therapeutics and worked to meet the FDA requirements for approval. Luca explained that approval required a great deal of work on the process, the analytics, the quality system, supply chain, and raw materials. One of the most transversal aspects of the validation of a product is getting it ready to be manufactured for the market and it was great to go this last mile with a strong partner like Orchard Therapeutics. He also credits the infrastructure of AGC Biologics, which is a multi-site global organization and provided the Milan site great support in terms of general quality, standards, procedures, and simply by having faced multiple FDA inspections before. The combination of all these factors was what carried them to the finish line, it required extensive teamwork, not only at the Milan site but also the entire organization. The Only Site to Receive EC and FDA Commercial Manufacture Approval I followed up by mentioning how with approval from the European Commission and the FDA, the AGC Biologics Milan facility is the only one in the cell and gene therapy industry to have commercial manufacturing authorization from both the FDA and the EC for LVV and cells. I asked Luca why there are so few CDMO's who have achieved this and what makes the FDA and EMA approval process so challenging? He explained that the Milan site was the first site to receive clinical manufacturing approval for an ex vivo gene therapy in 2003, 21 years ago, when cell and gene therapy was almost nonexistent at the time. They were the first facility to receive approval for commercial manufacturing in Europe for a marketed product in 2015/2016, 10 years ago. Now they are the only site who can do viral vector and cell therapy, both approved from the main authorities, the EMA and FDA. Most of the time, different services are provided by separate facilities, but AGC Biologics handles everything internally. This approach dates back 28 years when market capacity for such processes was lacking, prompting AGC to build its internal capabilities for vector and cell supply across all geographies. They continue to benefit from their 30 years of accumulated experience. Few Contract Manufacturing Organizations (CMOs) are commercially authorized, and none have been authorized for a decade and for multiple products. AGC Biologics is the excess manufacturer for Strimvelis and has four other filings under review (two with the EMA and two with the FDA). Additionally, they have completed three PPQ filings that are expected to be reviewed in late 2024 or early 2025. Luca explained that there are currently only 15-20 approved cell and gene therapy products, and AGC Biologics manufactures 5-7 of them, handling about 30% of all late-stage and commercially approved assets. This is due to the newness and difficulty of cell and gene therapy, meaning only pioneering CDMOs have reached this stage. AGC Biologics is proud of their position, and customers continue to rely on them because they offer early innovation, strong manufacturing capacity for late stages, and the quality needed for commercial approval. Keys to a Strong Partnership Next I asked Luca what he thought made the partnership with Orchard Therapeutics so successful. He emphasized that mutual trust and transparency have been crucial. They began collaborating with Orchard Therapeutics around 2017-2018, building daily on transparency, technical expertise, and science. Orchard Therapeutics owns the clinical portion and the product but relies on AGC Biologics for the manufacturing of the viral vector and cell therapy. They work together as an integrated value chain, fitting perfectly into the process of collecting cells from a patient, processing them, and then returning them to patients. Their partnership operates as a single team, continually innovating new solutions and processes. Transparency is key, with a "bad news first" policy to openly discuss any issues with their customers and find solutions together. This approach led to a realistic and ambitious plan that ensured they were well-prepared for regulatory inspections, resulting in their FDA approval. They aim to leverage this success for future approvals and product developments to treat more patients in need. Advice for Developers in Cell and Gene Therapy Regarding advice for developers working on cell and gene therapy products, Luca highlighted the importance of being realistic and pragmatic. Balancing the life-saving potential of these therapies with the need to maintain quality and effectiveness is crucial. A conservative approach might not be viable economically or too slow to meet the needs of the therapy. Developers should rely on experienced partners who have demonstrated credibility in the market. The field has seen significant ups and downs, often driven by unmet expectations and poorly executed ventures. Many new CDMOs have emerged, some overpromising on timelines and costs. Some developers may have been thinking that cell therapy is easier than it is, or simply a copy and paste of other modalities and some CDMOs found themselves having extra capacity and therefore bid aggressively with timelines and cheaper prices. This has set back progress, underlining the importance of realistic expectations and partnering with trusted, experienced organizations to navigate the complexities of cell and gene therapy development successfully. Therefore partnerships are essential, not just with CDMOs but with all players in the value chain who have proven their credibility. The Future for AGC Biologics I also asked what the future holds for AGC Biologics. Luca explained that in the coming year, they anticipate manufacturing 7-8 commercially approved products, significantly impacting the industry. Despite this growth, their focus remains on innovation and continuing to serve their early-stage customers, including academia, startups, and mid-sized biotechs in preclinical to Phase 1-2 stages. They are continuously innovating their proven processes in gene therapy (lentiviral and retroviral vectors) and cell therapy (stem cells and T cells) and are expanding their capabilities in AAV (adeno-associated virus) for in vivo gene therapy and investing in emerging cell types like exosomes, T regulatory and others. They enjoy partnership and bringing their customers into commercial manufacturing, but they always start very early in the process, and this is where their innovation is crucial. Milan develops the platform processes and then transfers these technologies to their other sites in Colorado, USA, and their upcoming site in Yokohama, Japan. AGC aims to establish manufacturing sites for cell therapy across three geographies (Europe, US, Japan) and for viral vectors in Europe and the US, with Milan as the innovation center. This strategy enhances their platform offerings and geographical reach, driving forward their comprehensive and strategic approach in the industry. I closed the interview by asking if Luca had anything else he would like to add. He emphasized that AGC is more than just a manufacturer; they are a Contract Development Organization (CDO) focused on development. While manufacturing is a significant part of their operations and they excel at it, their true value lies in their development capabilities. They can work with mature processes, adding value through suggestions, modifications, and scaling up. However, their strength is supporting clients with less mature processes. With 28 years of experience primarily in development, they have created processes from scratch for various cell types and developed comprehensive analytics. They have conducted over 160 tests, many of which are validated for market readiness. ...

In this podcast, we spoke with Nainesh Shah, Senior Application Engineer at Asahi Kasei Bioprocess about buffer management including the benefits of inline buffer formulation, and single use inline buffer formulation systems. Buffer Management We started the podcast by talking about how critical buffer management is to bioprocessing. Mr. Shah discussed how buffers are required in large quantities during the biomanufacturing process and that traditionally buffers were made in large tanks, stored, and used as needed. However, now real estate in the bioprocessing industry is at a premium and companies are looking to utilize new technologies that can reduce facility footprint. For buffer management, it makes sense to create buffer on demand to reduce the footprint dedicated to buffer production in the past. Inline buffer formulation is a hot topic with companies who require a large quantity of buffer because it provides a way to create buffer on demand in a much smaller footprint. The interesting thing is that it is now also a hot topic among small R&D scale buffer users as well. Inline buffer formulation systems are ideal for users who need 200 to 500 liters of buffer at a time. The system takes the concentrate and adds clean water to provide just the right amount of buffer on demand. Another benefit of inline buffer formulation is that you can achieve a quick process changeover and move on to the next buffer formulation without spending valuable time cleaning the tank, taking samples, and readjusting the critical parameters. Recently, any new manufacturer, whether it's a large scale or small scale tends to move into this field of buffer management and operates one or two Inline Buffer Formulation (IBF) systems like the MOTIV™. They then use these systems to make all sorts of buffers needed for their various processes. The MOTIV Family of Inline Buffer Formulation Systems Next, I asked Nainesh if he could talk a bit more about the MOTIV family of inline buffer formulation and fluid management systems that Asahi Kasei Bioprocess America (AKBA) offers. He explained how the award-winning MOTIV family has evolved into a series of inline buffer formulation systems designed to help companies move past downstream bottlenecks by driving buffer productivity. The product family includes 3-pump, 5-pump, and custom IBF configurations that can fit most any space, cost, or performance requirements. The MOTIV is a leader in buffer production with a range of scale from 4,500 liters per hour to 10 liters per minute to fit an entire range of volume requirements. He went on to say that they have added a new feature where MOTIV can fill up bags with buffer and monitor the quantity in the bag to make buffer on demand even easier. MOTIV SU Then we talked about the new MOTIV SU, a single use inline buffer formulation system, built to produce complex buffers on-demand effectively and efficiently, all from one pump head, and without the need for CIP/SIP procedures between batches. The innovative design modulates flow through control valves while simultaneously integrating buffer solutions and mixing. As with all the MOTIV systems, OCELOT System Control ensures precise blends every time, controlled by pH and conductivity feedback or flow. The MOTIV SU is perfect for a biomanufacturer who does not want to spend time with cleaning and validation. It is great for one time use as it does not require time spent in cleaning, validation, and making sure that it is free of all the contaminants and all the buffers which may be harmful for the next process. Another benefit would be if a biomanufacturer used a buffer which had a chemical or ingredient which would be problematic for other processes, and they wanted to eliminate any risk of contamination. Since the MOTIV SU has replaceable parts, which come as a pre-built unit, it is easy to replace the components and then the system is ready to run again. The MOTIV SU is on the same platform as the MOTIV and is an ideal solution for companies that don't want to disrupt their regular production lines, but still want to use the same production criteria that in the future could be added to production lines. Nainesh went on to talk about why single use is such an important option. He said that because there is so much time, energy, and money being spent to clean and provide the proper validation, which then sometimes needs to be reverified or revalidated later. Single use is a good option, and if the process permits, it can be used for a long duration of time provided you have the backup data that you are not contaminating. If you're using the same material repeatedly, yes, it's a single use, it's a replaceable part, but it doesn't mean that it's a one time use and then it is gone. It is just like the bioprocess industry using 50 liter and 100 liter bags for holding their buffers, which are all single use and this goes along that same line for the bioprocessing industry. Single use versus stainless steel Next he walked through when you would choose a single use option versus a stainless steel option. He explained that a primary consideration would be if you are using anything that would be considered corrosive. If a company is using something in their process and they are not sure whether it is corrosive and they don't have any documentation or any test reports to show whether it is corrosive or whether stainless steel will hold up and they don't want to damage the stainless steel equipment, it would be good to have a single use option as a fail proof. They can start bioprocessing studies in the single use system and at the same time start doing some studies with stainless steel to see if there is any issue with corrosion. In six months time, if they don't find any corrosion issues, they can switch over to a stainless steel option. Another reason to go with single use is color. Any chemist or bioprocessing person would know whether it's going to impart coloration or affect the stainless steel from the inside, that is again a quick answer to go for a single use option. Another area where single use is a good option is if you are considering whether to invest and want to prove that your technology works. You could start with the single use and then when you have a more permanent situation you can move into stainless steel. I followed up by asking if there are any striking differences between the single use and the stainless steel option. He explained that there are a few things that need to be considered when you're using a single use system. First, because it uses only one pump, which is also a mixer and it creates a suction force to take the concentrates in, there are limitations on the maximum or minimum flow that you can get and the concentrations. It is not good to use very concentrated material and then use a small quantity per minute to do the dilution. So that is a limitation that the user should consider. The MOTIV SU works on the same principle of mixing feedback control loop for the pH and the conductivity, or the flow and it can control in all the three parameters. It has its own tolerance values that the user can put in of what is acceptable or not acceptable. When it is not acceptable, the system will divert buffer out to a waste outlet and then when it gets back into specification it will revert the buffer back into the product outlet or the designated outlet. Collection of the buffer will start and the system will monitor how much is collected. The user can define and set all these parameters in their method, which are very easy to use and intuitive, just a quick 15-20 minute walk through can get a team up and running. Set up and operation Next, I asked if he could talk specifically about the operating system for this unit and also answer the question of if an end user wants to create a new buffer, do they need to come to AKBA. He said that the operating system is based on a Windows based PC housed inside the electrical enclosure with Allen Bradley controls for all the control mechanisms. The user does not have to contact AKBA or work with AKBA if they want to add any new buffer or modify a new buffer. They can make their own buffers, but we teach the user how to make methods via a drop down menu with fill in blanks. Then they can run the method and the system is ready to go. Customers can always come to AKBA if they're having any issues with making a buffer and we are always there to help guide them through. Lastly, I asked what the set up is like for end users who are interested in implementing the system. He explained that the regular MOTIV and the MOTIV SU are all skid units which are mounted on wheels that can be easily wheeled in and out of an area as needed. As he said earlier, real estate is very important in the bioprocessing industry and rooms are being utilized for multiple purposes. The mobility of the MOTIV family means that the user can connect to an electrical power supply that can be 110, 220 or 230 volts, in some instances with a higher flow rate it may need the three-phase power supply, but those are all plug ins. Then they connect the air supply for the valves and everything to open. Then connect the concentrate containers, that can be bags, 1 meter cube totes, drums, etc. Lastly, they direct where they want the buffer to go whether it's going to a container sitting right next to the system, or it could go into a port that can transfer the buffer into another room. That’s it, very easy and no special tools required for starting the system up or shutting it down. The floor size requirement is a typical office table space. It has a small HMI on top of it to see how the process is moving and all the pertinent data is is displayed there. If there are alarms it will show the user right there what is happening and the operator or the user can take care of it right away For more information,...

In this podcast, we talked with Dr. Ma Sha, Head of Bioprocess Applications at Eppendorf SE about advancements and challenges in cell and gene therapy production along with solutions for scale up and transition to stirred-tank bioreactor suspension culture. We began the interview by talking about the biggest advancements in cell and gene therapy, including CAR T-cell therapy development, clinical results and FDA approvals. Another area of great advancement is induced Pluripotent Stem Cell (iPSC) Culture technique. Dr. Sha explained that it used to be very difficult to culture iPSCs until it was possible to culture iPSC suspension spheres in stirred-tank bioreactors, which was a big breakthrough in the cell and gene therapy area. I followed up by asking Dr. Sha what he sees as the major challenges in the development and production of cell and gene therapies that still need to be addressed. He said that one of the major breakthroughs has been the autologous therapies that have been approved, particularly CAR T-cell therapies. However, this has also been a major challenge, because the autologous model is not cost effective. As a result, there has been a shift toward developing allogeneic therapies and building this production model will be a major challenge moving forward. Another challenge on the manufacturing side is ensuring to follow Good Manufacturing Practice (GMP), as it has been a common request from cell and gene therapy companies. Next, I asked him about the move from 2D to 3D culture and his experiences with this transition. Dr. Sha shared that several of the projects that Eppendorf bioprocess works on start as 2D culture in flasks, it is a natural place to start for most of the cell lines since they are attachment cells. They must then be converted into suspension culture to enable 3D culture, since 2D culture significantly limits the yield and productivity. He went on to say that if you look back at the evolution of antibody production, it was important to convert production to suspension cell culture and this is also necessary for the cell and gene therapy field. Moving from 2D to 3D culture and especially utilizing stirred-tank bioreactors enables much higher yields. As it stands, the yield for cellular therapy cell production is fairly low, especially compared to the industry standard of CHO cells used for antibody production, so a lot of improvement needs to happen. We then discussed stirred-tank bioreactors and their increased use in cell and gene therapy development and production. I asked Dr. Sha what are the key factors that developers should consider when choosing stirred-tank bioreactors. He explained that stirred-tank bioreactors fit the model of allogeneic production. Autologous models are not suitable for stirred-tank bioreactors. Developer companies need to keep in mind that if they want to move to stirred tank bioreactor platform, they need the production model to be allogeneic. In addition, it is important to consider the support available with respect to scaling up and leveraging supplier experience. For example, Eppendorf bioprocess over the years has produced many application notes to help customers scale their manufacturing. They have even built model production systems in Eppendorf stirred-tank bioreactors. The program called “Scale up Assist” allows customers to skip much of the difficult calculations required to achieve reproducible yields when moving from smaller to larger vessels. Eppendorf has a very long history of working in protein-based therapeutic cell culture production and about ten years ago expanded to include cell and gene therapy. I asked Dr. Sha in his experiences, what are the most important takeaways in terms of areas that still need work and advancements on the horizon. For instance, what can we learn from protein-based therapy cell culture to apply to cell and gene therapy production? He said that he thought that allogeneic production is a great lesson learned because simplicity is king. The ability to produce one therapy for different patients from one source cell line would greatly simplify the workflow and cost model. He stressed that the one takeaway that must happen for the field to become more successful would be the allogeneic model. This model also fits perfectly with stirred-tank bioreactors. Another takeaway for stem cell therapy would be the need for more success stories, including more FDA approvals to build successful examples so companies can follow the same path. There are a lot of cell therapies in clinical trials, but the number of approved therapies is small, especially when compared to CAR T-cell therapy, for example. I closed the interview by asking Dr. Sha if he had anything else to add. He said that in addition to Eppendorf’s long history with stirred-tank bioreactors, they have acquired two bioprocess companies previously known as New Brunswick Scientific and DASGIP GmbH to add to their significant history and expertise in bioprocessing. In addition, Eppendorf has a lab in the United States and one in Germany where they are developing sophisticated cell and gene therapy applications including iPSC suspension culture in their SciVario® twin bioprocess controller. Using this tool, they have developed a very high-density mesenchymal stem cell culture of around half a million cells to 1,000,000 cells per ml. In an upcoming application note, they will showcase their extremely high yield 15 million cells per ml bioreactor solution. Eppendorf is committed to developing cutting edge applications to support customers in the cell and gene therapy space. To learn more, please see: Eppendorf Bioprocess Application notes links: Stem Cell Exosome Production on the SciVario® twin, a Flexible Controller for Your Bioprocess Needs The DASbox ® Mini Bioreactor System as a Tool for Process Development And Stem-Cell Derived Exosome Production in Standardized Culture Conditions

In this podcast, we talked with Dennis Hodgson and Phil Sanders from Agilitech about the benefits of single-use mixers, dealing with supply chain concerns, ensuring scalability, and tailoring a mixer to meet specific process needs. Benefits of Single-use Mixers We began the podcast by talking about the overall benefits of single-use technologies for mixing. Dennis explained that single-use mixers are very versatile and can be used to replace stainless steel vessels within the manufacturing area. Single-use mixers all have the same advantages of other single use components, such as coming fully sterile and eliminating the need to steam and clean in place. Dennis went on to say that another big advantage that single-use mixers have over stainless steel is the ability to customize. For example, a 500 L single-use mixer can be used with a virtually unlimited array of customized vessel configurations, which would include the inlet outlet, port configurations, sampling ports, vent filters, and various process analytics that can be added. Next, we talked about adoption of single-use technology for mixing and possible concerns that customers might have. Dennis shared that a big concern recently has been supply chain shortages that have created limited availability and long lead times for single-use consumables. He said that he has heard from some clients that they have had to skip planned production batches because the single use bags that they needed to process the batch were not available. Phil added that supply chain concerns have caused some of their clients to think about moving to stainless steel systems to avoid any production delays. Single-use Technologies Supply Chain Challenges I followed up by asking what could be done to address single use supply chain issues moving forward. Dennis explained that Agilitech has the luxury of not being tied to any one supplier, so they can source from multiple vendors. This allows them the flexibility to move between vendors and load projects based on their capacity and lead times. This also allows them to make sure that they are offering competitive pricing because vendors know that they're not the sole source of a component. Ensuring Flexibility in Single-use Mixing We then talked about mixers presenting unique challenges in that they are used for a variety of applications with many different demands. I asked how Agilitech can ensure that their single-use mixer has the flexibility needed for multiple applications. Dennis explained that because Agilitech isn’t tied to a single design, they are able to have conversations with the client to customize a solution for their needs. Their main goal is to make a product that meets the needs of the individual companies and their process. Additionally, they design their systems purposefully to handle many different capabilities such as sampling, analytical measurements, weight measurements, temperature control, etc. Because they use standard control hardware, their mixing vessels can easily be integrated into existing control systems such as Delta V or Wonderware through the available Ethernet IP connection. This allows users to read and write to certain control parameters. I then asked about which options are available for customization on the single-use mixers. Dennis said that they can customize all the inlet and outlet ports with regards to port size, tubing length, connector type, etc. As far as the mixing units themselves go, they can be jacketed or not, have load cells or not, have probe analytics such as pH, conductivity, temperature, DOE, and optical density, so all those different analytical devices can be incorporated as well. Phil added that if there are specific standards within an organization, for what control systems need to be installed on these systems Agilitech is flexible with Rockwell, Delta V, Siemens, all the major platforms that customers might need. I followed up by asking about how these customization options affect the cost of these mixer systems. Phil said that frequently when customers hear the word customization or even tailored, they think that is going to drive the price up or maybe even drive the delivery time out. When Agilitech tailors a solution, it speeds up the delivery time and reduces the cost because everyone is on the same page and has alignment on what will be delivered. Agilitech works through all the issues up front with clients, so customers get exactly what they’ve asked for. Single-use Mixer Scalability We then discussed scalability and about how low volume mixers are difficult to find. I asked how Agilitech has been able to build scalability in their product line. Dennis described that their current sizes that range from 10 L to 650 L. He shared that their 10 L size has a minimum working volume of 1.25 L, which permits a process application such as a highly concentrated TF step to be able to be used with that mixing vessel. Additionally, because Agilitech has a total of seven different sizes, it allows customers to scale up their process for additional capacity using identically designed equipment. Levitronix is working on motors and impeller sizes that would permit Agilitech to offer up to a 3,000 L size in the future. Easy Process Implementation Next, I asked Dennis if he could talk about process implementation. He explained that each vessel comes with its own easy to use touchscreen interface that allows users to locally control the start and stop of the motor, change the setpoint of the speed on the agitator, and read local measurements for the weight, temperature, and other process analytics that are connected. Also, by using standard industrial hardware and standardizing on up-to-date future proof communication protocols such as Ethernet IP these systems can very easily be integrated into existing control systems. Lastly, I closed the interview by asking Dennis and Phil if they had anything else that they wanted to share with listeners. Dennis said that the industry has gotten too used to there being too few options in the market and perhaps have started to shape their processes to align with what is currently available. At Agilitech, the aim is to shift that perception and put the power back with the end user to define their equipment requirements that meet their process instead of vice versa. For example, changing the inlet and outlet size of your single-use mixing bag to better fit your process shouldn't increase the costs and time to receive that bag significantly. End users should remain rigid in their requirements and process. Vendors should be the ones that remain flexible to meet the needs of the user. Phil added that the solution should be plug and play to some extent. It shouldn't be a monumental effort to implement a new solution in an existing process. A plug and play approach allows companies to get to market faster and to source components quicker. To learn more, please visit Agilitech Single-Use Mixers.

In this podcast, we spoke with Cory Hinz, Engineering Manager at Asahi Kasei Bioprocess about the different methods that are available for liquid chromatography mobile phase solutions and the benefits of inline blending. Cory also describes how to implement binary blending feeding of a liquid chromatography process using inline blending. Liquid Chromatography Mobile Phase Solutions I began the discussion by asking Cory if he could tell listeners about the different methods that are available for liquid chromatography mobile phase solutions. He explained that for chromatography, it's important to remember that the process and the chemistry should drive the method used. Some chromatography processes use prepared mobile phase solutions that don't require inline mixing, while others blend two or more solutions together to formulate. The mobile phase takes these blends and changes their composition over the course of the elution. Each of these methods is driven by the needs of the process. Inline Blending Next, I asked Cory if he could tell us about the benefits of inline blending. He said that inline blending allows solutions to be prepared at their point of use, not just for chromatography processes, but any blending process. This increases the consistency of the blended solution, reduces dependency on the accuracy of raw materials, allows for real time quality assurance, and eliminates the risks and extra resources and space required for traditional tank approaches. Inline blending also adds an element of flexibility, allowing functions such as buffer preparation to become more of a utility than an additional process. Binary Blending Feeding of a Liquid Chromatography Process using Inline Blending Cory then provided details about how to implement a specific solution for binary blending feeding of a liquid chromatography process using inline blending. He explained that binary blending is the most common configuration they see for their chromatography equipment customers, because medium and high-pressure liquid chromatography require a dedicated pump to supply the pressure dictated by the process. It is important to design the binary blending at the suction side of the pump. This is done by employing two modulating control valves, one for each of the two components of the mobile phase, and ensuring sufficient supply pressure to each one. He then told us about the role that each of the valves play in creating the ideal blend. He described how the control valves do most of the heavy lifting for binary blending. The first valve controls the diluent, which will be the purified water or buffer that comprises the majority of the mobile phase blend. The second valve controls the component that is getting diluted. These valves each react to a different process parameter to achieve high accuracy. The second valve is the most intuitive, the component being diluted can have its proportion increase or decrease based on the movement of the control valve. For example, if the concentrate is below target, the valve will open to allow more concentrate through. This can be based on flow connectivity or any critical process parameter that can be measured inline. The first valve is less intuitive. It is controlled by the pressure in the system after the two streams have combined. If the blending pressure is too low, for example, the valve will open to increase that pressure. The result of this configuration is that if the two valves react to one another via the process but are not linked by a system control algorithm. This results in flexibility and accuracy and also provides a way to monitor and mitigate pump cavitation. Next I asked Cory about controlling the incoming process pressures of each of the valves. He said that in order for the binary blending scheme to work optimally, the incoming supply pressures of each stream should be controlled to prevent fluctuation that can disrupt the automatic blending control. To accomplish this, they employ a booster module on each of the two inlet streams. They have found that this additional feature is successful in isolating the system from fluctuations in supply, due to facility or other environmental effects. He went on to say that it is also convenient to match the two supply pressures to one another because that creates an identical pressure drop over both belts and a more intuitive blending scheme. He then explained how the booster modules work together to maintain pressure. The booster modules are identical and are each composed of a recirculation loop driven by a load pump. Once primed, the load pump runs at a constant speed and directs flow both through the recirculation circuit and to the control valve. By placing an aero augmented back pressure regulator on the recirculation line, we can automatically control the effective pump discharge pressure. This is the supply pressure that each valve sees. The air augmented design of the back pressure regulator means that the system can target the desired pressure set point by modulating the supply air to the regulator. I closed our session by asking Cory about other formulation challenges that he gets asked about. He said that not all applications require binary blending, some require combinations of three or more components. This makes system control more complex but is a familiar application for Asahi Kasei Bioprocess. They have tackled this several times in liquid chromatography applications, but the most common implementation of multi component blending is for buffer formulation. Their line of MOTIV™ systems is specifically designed to achieve highly accurate multi-component blends using our proprietary Pro-Yield™ technology. Both their chromatography and MOTIV formulation systems operate on their state-of-the-art, industry 4.0 process control software known as OCELOT™ System Control. LEARN MORE ABOUT AKBA’S BIOPROCESS CHROM SYSTEMS

In this podcast we spoke with Klaus Kienle, Global Product Manager for the Mixing portfolio at Pall Corporation about the latest mixing technologies including single-use solutions, the need for increased flexibility, and a more standard vendor agnostic approach. The Role of Mixing in Biomanufacturing I started the conversation by asking Klaus if he could talk about the role that mixing plays in biomanufacturing and current challenges in this area. Klaus explained that mixing is an omnipresent process. It starts with upstream buffer media and ends in fill and finish. It is an important part of manufacturing across several modalities, including monoclonal antibodies, mRNA based vaccines, gene therapies and various other processes. Across these various applications, flexibility is key, and it is also the primary challenge for the future. He continued by saying that Pall customers have expressed that they want increased flexibility, better lead times, and less supplier dependency in the future. Advancements in Mixing Next, I asked about the latest technological advancements in mixing. Klaus stated that the latest advancements are moving towards tackling the flexibility challenge, which means supplying solutions that are available with shorter lead times and are more vendor agnostic, so they fit with other vendors’ manifolds and full sets. This is consistent with the recent launch of the Allegro™ Ready Standard Solutions from Pall, which is not only limited to mixers, but also includes storage transfer sets and other segments. Pall has launched this new standard set ranging from a 30 liter mixer up to a 3000 liter mixer. I continued the discussion by asking if he could talk a little bit more about some of the additional advantages of this set of new standards. He described how these standards are ready to go, so if a customer is asking for a manifold, there is no time required to generate a drawing or waiting for pricing back, resulting in a short lead time. Pall is working towards having these standards available off the shelf, reducing lead time further with availability in the range of single digit weeks, depending on manufacturing and where the customer is located. He shared that Pall has invested $1.5 billion to increase capacity and reduce lead times. Single Use Mixing I then asked him about some of the remaining challenges that exist with single use technologies. Klaus explained that one of the main challenges that the remains in single use is sustainability, especially since it consists predominantly of plastic components. However, there was a recent publication in New Biotechnology, authored by biopharmaceutical companies, “Streamlined life cycle assessment of single-use technologies in biopharmaceutical manufacture.” It makes the case that single use technology is providing better sustainability in the biopharmaceutical process because single use technology allows customers to use more intensified processes, thereby increasing the efficiency per consumable. Single use technologies also support a closed process and reduced clean room requirements resulting in lower energy requirements. This is in line with the new standard designs from Pall, where the filtered product line is fully closed and processing ready. For instance, now the powder port on these designs ensures a closed and controlled environment. This then allows bioprocessing companies to lower the cleanroom environment requirements, which translates to significant energy savings. Modular Mixing Approach I continued our discussion by asking Klaus about a recent white paper published by Pall and Lonza, that highlighted a modular mixing approach. (need paper link) He described how a modular approach can give customers the flexibility that they are looking for to adapt to new requests, especially in the contract manufacturing organization (CMO) environment. Global CMOs are producing product for developers and as a result, they are frequently changing the product they are manufacturing. Their facility needs to be set up with the greatest degree of flexibility to adapt to changing needs as well as short notice production demands. For example, the need to be able to respond under short notice was clearly seen in the push to produce the COVID-19 vaccine. If a CMO gets this request and they’ve built their facility with a modular approach, systems could be interchanged easily. This would allow them to adapt quickly from a monoclonal antibody process to an mRNA process. A modular approach can also save companies on CAPEX as some consumables they will have in stock. The paper illustrates the ease of use of Pall mixer systems and their flexibility, which allowed a fast change over without buying new equipment. What to Consider when Selecting a Mixing System Next Klaus provided advice for listeners about what end users should be considering when selecting a mixing system. He replied that there is a message right now from some suppliers that one size fits all solutions are the best option, but the reality is simply that most processes are so unique and so variable that one size fits all doesn’t work. Pall takes a different approach by offering four different mixers providing a variety of solutions depending on the process. For example, if a customer needs a very powerful mixer, Pall offers the magnetic mixer, but if they need a shear sensitive one, they recommend the LevMixer® system. Because they utilize the same mixing tank, a simple change of the drive unit makes the equipment ready for either the high power demand of the magnetic mixer or the shear sensitive mixing of the LevMixer system. This flexibility allows customers to quickly adapt their entire facility towards a specific process without exchanging the big capital systems. Before we wrapped up the interview, I asked Klaus if he had anything else that he'd like to add for listeners. He told me that the Danaher philosophy is listening to the voice of the customer. Pall embraces this and takes it very seriously. This means that Pall reaches out to customers to ask what their specific needs are and how they can improve. One key slogan within Danaher is continuous improvement and Pall has applied that to their product portfolio by continuing to work on providing flexibility and interchangeability, not just within Pall, but future interchangeability between several technologies in the market. He added that his personal goal is to deliver the next level mixing experience, which in some cases is not the next level technology. It might be the next level of flexibility instead of the technology, but in the end, the bottom line is that the customer is experiencing happy mixing. For more information, please see the white paper Trends in Single-Use Mixing Technologies for Biomanufacturing

In this podcast we spoke with Derrick Alig, North American Western Regional Sales Manager for PSG Dover Biotech, Chris Couper, President and Founder of Liquidyne Process Technologies, and Phil Sanders Biotech Chief Innovation Officer at Agilitech about current supply chain challenges, possible solutions, what the future holds, and ways to navigate supply chain shortages to ensure manufacturers meet their timelines. Supply Chain Shortages I began the discussion by asking our panel members if they could discuss challenges that their customers are currently having with sourcing single use consumables and technologies. Derek began by discussing the lack of raw materials to make these products, whether it is polymer-based components where lead times have been extended due to lack of raw materials, or other areas such as chips. As a result, customers are having to purchase larger quantities of product in advance, which ultimately leads to even longer lead times. Chris added that from a distribution perspective and an integrator perspective, many of their primary suppliers have had issues. They have also seen that many manufacturers have been able to ramp up their production with plants that were put in place in 2019-2020. However, it takes one to three years depending upon the complexity and the scope work to create additional manufacturing facilities and production lines. In addition, many manufacturers are using alternate materials. While they may have qualified one product in the past, now they are qualifying additional supply chains, so they have a primary supply chain and also secondary and tertiary chains. Phil discussed bringing an agnostic approach to managing supply chain shortages to alleviate some of the issues of having a single source. He also pointed out that sometimes the focus is on single use supply chain issues, but there are companies using reusable equipment that are having some of the same supply chain issues, especially when it comes to these things like chips and control systems. Supply Chain Solutions Next, I asked the panel how their individual companies are approaching these challenges, specifically how they are working with customers to provide solutions for these challenges. Derek explained that at PSG Dover they are committed to providing quality products to customers in the biotech market. They focus on delivery times for customers by adding more shifts to keep up with demand and in late 2021, they added a second validated cleanroom to provide additional production capacities. They have also acquired companies to provide additional capacity. Chris added that the situation could have been much worse if manufacturers had not stepped up and added capacity like PSG Dover and that they have seen improvements in lead times. He added that for Liquidyne they have a minimum of three supply chains for virtually every component that they offer. They let their customers know that they need to qualify the three components so they can be used interchangeably to meet timelines. Phil added that maintaining flexibility is critical. For example, maybe the entire amount of inventory that is needed isn’t available now, but there is enough to get started while orders are placed for the rest of the material. Instead of trying to provide a customer with inventory for an entire year, provide three months' worth, then another three, and so on. Supply Chain in the Future I followed up by asking what they thought the future looks like for the supply chain over the next three to five years. Does this resolve itself or does it shift to another potential supply chain shortage? Derek said that he thinks that customers will continue to require multiple supply chain solutions and suppliers will also need to continue to add multiple sources for their raw materials and electrical components. Chris said that he thinks companies that are successful will take the time to study what has occurred, how they reacted, what could have been done differently, potentially better, and what missteps should be avoided. One thing that we've learned as an industry is that we must be able to react quickly to the market, science had the ability to react quickly to this pandemic and they moved vaccines and treatments to the market in an incredibly short timeline. They took things that historically take several years to accomplish and did it in a matter of months. Suppliers need to be able to respond quickly to increased demand as well. Ultimately, he thinks that customers will need to communicate about what they need in for one to two years in advance instead of in three to nine months. Phil said that he thinks the biggest issues are what we don't know today, what geopolitical event is going to happen that could cause sanctions on a country where we're getting components from and then having to shift those things. What act of God, tsunami, or earthquake, etc. If we can start figuring out what issues that coming, maybe we can maintain a specific inventory on these components. Maybe Chris keeps some of them, Derek keeps some of them, and I keep some of them so that we're working together to provide solution. He thinks over the next three to five years, the things that we're talking about today are going to be a thing of the past and we're going to be talking about a new supply chain shortage. Chris added that as three companies we look at risk and try and share that risk to create a better overall customer experience. We must work together in order to make that a reality, and we have to make sure that we communicate individually and together as a team with the client base that exists along with the client base that we don't even know about yet. All the panelists agreed that there needs to be excellent communication and partnership with customers to fully understand what their needs are and to help them ask the right questions to ensure that they will have the supply they need to move forward. Navigating the Supply Chain Then the panel discussed the things that they recommend customer related to supply chain, not just for purchasing and production, but also for scientists. Derek started by saying that what they really want to know from their customers is their timeline so they can match it up with what they can deliver. It allows PSG Dover to review all the different options available for the best solution possible and also to make them aware of what is realistic with the current supply chain. Chris added that it is critical to know what the key components for a process are and what are the risk items or items that have volatile lead times. It is critical for customers to be able to rank these items in terms of risk. Because of volatility in the marketplace in terms of getting the product or volatility in terms of price, we're seeing price adjustments 30 days' notice and it's not only for future orders, but also for orders that haven't shipped. New price increases happen because the raw materials that were used to build the product have escalated in price, and this is what's being passed down the supply chain. It is very important to look at the high-risk items and make sure that the product is secure and that there isn’t an issue in a year or two. Phil explained that customers need to understand that assemblies are different components. So, you can't look at an entire assembly and say that that whole thing is at risk. It's the different components and understanding that sensors or pump heads or connectors or any of these things can be a supply chain issue and having that discussion with the team is important. It is important for customers to think about if there is an issue with one of the components, what is plan B? Is there inventory on the shelf or are there other options that can be explored. Phil then gave an example from one of their customers that had a specific design to be able to maintain their TMP control, but the issue was that the team wasn’t sure that the supply chain for that design could be maintained. Instead Agilitech built the unit with interchangeable parts, so if that a component wasn't available, there were others on the market that could be substituted. He reiterated that it is important to be adaptable and flexible and have those conversations, so you have a plan for managing supply chain issues in advance. Chris added that it is very important to have the entire team present when in discovery meetings with the customers. In order to facilitate quick turnaround times, these meetings need to involve not only the scientists, but ideally project managers, people from automation, and people from quality and documentation so that all the expectations can be addressed and met. This post is sponsored by Liquidyne Process Technologies, Inc.. This post is sponsored by Agilitech.

In this podcast, I spoke with John Ketz and Denis Kole about viral vector production, including current manufacturing challenges, navigating the road to commercialization, and successful scale up strategies. We began the interview by discussing the strides made in cell and gene therapy. Denis shared information about the several approved therapies and the more than 2,000 ongoing clinical trials. Denis added that while the immense potential of these personalized therapies is becoming more and more clear, the challenges and bottlenecks surrounding their development and manufacturing are also becoming a reality. This is especially true when it comes to producing and delivering sufficient amounts of these complex therapies in a reasonable timeline. He explained that patient demands are increasing and the increase in IND applications for cell and gene therapies is resulting in increased demand as well as competition for resources. Access to qualified labor has become a significant bottleneck and likely will continue to remain so, at least for the foreseeable near future. He went on to say that the lack of standardized approaches for gene therapy modalities is another challenge that can increase the risk of failure. It can result in increased process complexity due to the need to screen large numbers of variables and can result in extended development times, which in turn affect the time to market for these needed therapeutics. In addition, the availability of manufacturing capacity is also becoming quite limited with the field currently reporting a significant backlog that can extend as much as 16 to 18 months. John added that he also sees capacity issues and it is something that Andelyn Biosciences® is trying to address. He also said that they are working on increasing speed and consistency. There are a lot of challenges that need to be addressed when moving from small flask or bench top scale into larger production scales. These may be challenges that you are not aware of or don't encounter at small scale. It is important to be mindful of this during scale up. Addressing Current Manufacturing Challenges To continue the discussion, I asked them to share the best way to address these challenges moving forward. Denis began by sharing that the demand for clinical and commercial manufacturing for advanced therapies is expected to continue to increase as more drug development companies entering the space and new therapies continuing their development journey. As a result, the manufacturing backlog currently observed will likely continue to remain present, if not expand. So, while large pharma and a few larger biotech companies may have the resources to internally support their clinical development and manufacturing needs, most of the smaller and medium sized drug development companies will continue to face challenges associated with quick access to qualified labor and access to extensive process expertise and available manufacturing capacity. He went on to say that this is where groups like Pall’s AcelleratorSM process development services and Andelyn Biosciences can play an important role with support through partnerships and collaborations. These types of collaborations really can provide the necessary resources and support for therapy developers to target shrinking the development timelines, reducing the risks associated with process development, and scaling to commercial scale. In addition, Pall and Danaher's integrated single use bioprocess offerings, provide scalable solutions that can reduce some of the risks associated with the development and manufacturing of therapeutics. These solutions really aim to alleviate some of the bottlenecks and risks associated with the lack of a standardized approach. John added that at Andelyn Biosciences, they focus on creating a robust small scale model to ensure the consistency of the product when they move into larger scale. By doing this they can try to account for any of the differences that they might have, and can identify and define all of the critical process parameters throughout the whole process. Then they test the limits of all of these parameters in their model creation. He shared that working with a group like Pall, they have more consistency and confidence in the platform, so when they scale up, they know that it can work the first time and they don't spend multiple rounds on large scale experimentation. John continued by saying that to address capacity issues, Andelyn is currently building a new manufacturing facility that will be able to handle additional large scale manufacturing for gene therapies that are coming online. Choosing a Scale Up Platform I then asked them if they could talk a little bit about the key factors that developers should consider in deciding which is the right technology for scale up. Denis replied that where the industry stands today, it is important to realize that there's currently no one size fits all approach when it comes to production platforms and production systems for gene therapies. There are a multitude of factors that cell and gene therapy developers must consider when deciding what is the right technology and platform for scale up and it is imperative to evaluate the target patient population size, the overall product demand, and the timelines that are associated with the clinical trial milestone. This work needs to be done in parallel with assessing the risks associated with each development and production platform and evaluating the advantages and challenges associated with each solution. He said that he also believes that it is very important to think about a therapy’s commercial scale needs as early as possible in the development process. Starting this exercise early on, offers an opportunity to think about scalable manufacturing solutions early in the development process, and will likely lead to better process understanding and simplified transfer of the process to a manufacturing scale that meets the program's needs. John added that it is imperative to have confidence in all the small bench scale studies to be able to translate those results into large scale production. It is helpful to work with a company like Pall to aid in the backlog of experiments and increased manufacturing demands by limiting a lot of the time spent on large scale experiments on your product. Scaling up an AAV Manufacturing Process to 500 Liters Next, we talked about their upcoming webinar on how they scaled up an AAV manufacturing process to 500 liters. I asked them to share some of the highlights. John told me that he will be presenting how Andelyn used design of experiment studies for creation of a model that is robust enough at small flask and bench scale level to translate all of those optimized parameters into a 50 liter pool bioreactor. He will go into detail on their statistical approach and some of the results. He will also touch on some of the issues that they saw after moving from a small flask and benchtop into a 50 liter bioreactor. He will also discuss how they tech transferred that platform to Pall to help scale into a 500 liter bioreactor. Denis spoke next saying that optimizing and successfully scaling a viral vector process to manufacturing scale is not a trivial process. It requires extensive process knowledge and expertise, and one of the major hurdles is scaling the process while maintaining good process control, demonstrating equivalent productivity and maintaining product quality attributes. During the webinar, Denis will discuss how Pall’s Accellerator process development services team can support gene therapy development companies and then he’ll dive deeper into the specific approach that they took during the collaboration with Andelyn Biosciences. He will share some data that demonstrates the successful process scalability from the 50 liter scale to the 500 liter scale using the Pall Allegro™ bioreactor platform. The results demonstrated comparable viral vector production as well as comparable recovery between the two different scales, again using some downstream technologies like Pall’s Stax™ filtration platform. I closed the interview by asking them if there was anything else that they would like to add for listeners. Denis said that we are living in really exciting times, the gene therapy journey through process development as well as industrialization is really a complex one, but the benefits of successful development and robust commercial processes are huge. We are all witness to the extraordinary impact that development and manufacturing of these therapies has on patients. John then said this is a very mission focused project for Andelyn, and their goal is always patient focused and patient oriented - that's their #1 mission. All of Andelyn’s work has the end goal of improving people's lives and it's exciting to be a part of projects that are helping to drive the field forward. To view the webinar, please register at Successful Scale Up of a High Yielding Viral Vector Suspension Platform to 500 L