by Nitin Kulkarni, Senior Field Application Scientist, MaxCyte, Inc.
Cell and gene therapies (CGTs) are transforming the treatment landscape across oncology and rare diseases, enabling targeted approaches that are difficult or impossible to achieve with conventional small molecule drugs and immunotherapeutics.1 The sector is rapidly evolving, with advances in gene editing and cell engineering leading to an explosion in the number of therapies in clinical trials and moving to routine clinical use. This growth is shifting attention across the sector from questions around basic scientific feasibility to those about the realities of commercial manufacturing. A key challenge for developers of CGTs remains the transition from research to manufacturing scale-up, and the smoothness of this transfer can make or break new therapies.
The highly personalized nature of CGTs creates significant complications for the manufacturing process, leading to difficulties that are not present for conventional therapeutics. CGTs are often derived from the patient’s own cells, meaning each individual treatment requires unique handling, introducing significant variabilities throughout development and manufacturing. Protocols that show success during R&D stages often require significant modifications – or even replacement – during scale-up, adding time to the process, driving up costs, and potentially delaying regulatory approval and release to the market. Ideally, manufacturers should be able to trust that technologies used for cell engineering during development can maintain performance, consistency and reproducibility along the pipeline. However, achieving this is often challenging, not least due to differences between patients and the inherent biological variability observed within primary cell populations over time.
The value of consistency
Different challenges are introduced as CGTs move through development stages, from early research to proof-of-concept, through process development, scale-up and finally clinical manufacturing. In early research workflows, systems are typically optimized for flexibility and speed as researchers work to screen potential treatments. At later stages, processes must be adapted to the practical realities of larger-scale manufacturing (Fig. 1). Any changes in equipment, materials or process parameters as development is scaled up can introduce variability that requires additional optimization and validation, creating more pressures for development teams.2,3
Process reoptimization when scaling often introduces additional hurdles which extend development timelines, and potentially increase the risk of process failure even further. Variability introduced during the scaling process can affect the critical attributes of cell viability, transfection efficiency, or product consistency. Process changes also attract the scrutiny of regulators, particularly as programs move into clinical and commercial phases, with key guidelines such as ICH Q5E requiring manufacturers to demonstrate that reoptimization does not impact quality, safety, or efficacy.4 Together, these additional burdens highlight the importance of approaches that enable continuity between R&D and manufacturing to make scaling up more predictable and efficient. However, continuity is not possible for every technology or process: cells respond differently to changes in culture format, density, handling, and the microenvironment.5 Ideal strategies must account for this biological variability alongside consideration of engineering requirements, to allow reliable and robust manufacturing.
Considerations when scaling electroporation approaches
One example of a critical stage to consider when scaling is electroporation, a central technique applied in many CGT workflows. Electroporation uses brief electrical pulses to induce temporary permeability in the cell membrane, through which DNA, RNA and gene-editing components can enter without adversely impacting cell viability. Electroporation offers an attractive alternative to viral-based delivery systems, avoiding many of the limitations and safety considerations. However, despite the advantages of electroporation over viral vector delivery systems, it is not an out-of-the-box solution. Performance is influenced by cell type, buffer composition, process conditions, and the parameters set on the device. Therefore, users must carefully optimize settings to achieve delivery without impairing cell viability, and this will potentially require adjustment as processes scale up. Electroporation sits in the middle of the CGT development process, meaning its efficiency is impacted by what comes before. During R&D workflows, cells typically move directly into electroporation but, during commercial manufacturing, cells will often pass through longer fluid paths which lengthens their time in buffer and exposure to shear stress, oxygen and heat, increasing the potential for clumping. Larger scale processes are typically batched, which can exacerbate these problems. Together, these challenges can impact cell health and overall process efficiency, making it difficult for developers to maintain consistent performance when transitioning to scaled-up manufacturing.6,7
Established and proven scalable electroporation platforms can, however, help to reduce the disruption of moving between stages, improving the continuity of process conditions across development. These technologies enable entire protocols and parameters to be transferred with minimal modification, almost eliminating the need for repeated optimization. The seamless scaling process is only possible when using instruments and consumables that are well characterized for both electrical properties and the resulting heat generation, as these differ enormously across formats.
Mutually beneficial collaboration across the sector
The complexity of CGTs means that the right technologies and a significant level of technical expertise are needed during the development and scaling of processes. Optimization is often iterative, and requires input from specialists in both the biological system and in the technologies used in manufacturing. In many cases, historical datasets and established protocols can provide a reliable starting point for optimization, often considerably reducing the number of experimental iterations required, but many manufacturers – particularly start-ups and spin-outs – lack this data. Working with experienced commercial partners with significant experience and process knowledge across diverse CGT workflows can help developers across the industry to benefit from expertise.
The relationship between therapy developers and technology providers should extend beyond just the procurement and initial implementation of a new instrument; it should be a strategic partnership that delivers value throughout the development pathway. Engaging with commercial partners who routinely provide training, troubleshooting, and ongoing process guidance ensures that technologies are implemented consistently across different stages and sites, resulting in more consistent product production. Effective collaboration results in a shared understanding of process requirements and better long-term alignment between all parties. For developers, this reduces risk, improves reproducibility and smooths the progression from concept to clinic; for technology providers it acts as an opportunity to share expertise and gain an understanding of what collaborators want from technologies, leading to targeted improvements to products and systems.
Beyond technology, broader operational forces – including supply chain reliability, access to materials, and the availability of skilled personnel – can all influence the success of CGT manufacturing. Industry forums and collaborations are also crucial, encouraging knowledge transfer that can help to standardize processes across different sites and development stages. As CGT manufacturing expands, alignment between research, process development, and clinical production will become increasingly important.
Scaling for success
Ultimately, a complex set of technical and operational challenges shapes the journey of a new CGT from concept to clinic. As therapies move from early R&D stages to clinical use, it is essential to scale processes while maintaining consistency, quality, and regulatory compliance. Developers must strategically plan and engage across the industry in order to navigate these challenges successfully and avoid the need for reoptimization.
Electroporation – a central component of many CGT workflows – has a direct impact on both the efficiency of the process and the manufacturability of the end product. The ability to apply consistent electroporation conditions across developmental stages is essential for ensuring reproducibility and process continuity. Scalable platforms that maintain parameters across stages reduce complexity and improve overall reliability. Across the CGT landscape, successful deployment depends on the integration of appropriate technologies with technical expertise, often driven by collaboration between developers and technology providers. CGT manufacturing is an exciting, rapidly evolving field, and approaches that enable scalability alongside reliability will be key for advancing production of the next generation of therapies.
About the author
Nitin Kulkarni is a Senior Field Application Scientist at MaxCyte with more than 12 years of experience advancing cell and gene therapy applications from early-stage research through clinical development and commercial manufacturing. His expertise spans cell culture, molecular biology, immunology, bioprocessing, and microfluidic cell isolation technologies, enabling him to support customers across a wide range of therapeutic development programs. Prior to joining MaxCyte, Nitin conducted postdoctoral research in autoimmune diseases and cancer at Beth Israel Deaconess Medical Center and held scientific and product support roles at Corning Life Sciences and MicroMedicine. Drawing on experience across both academia and industry, Nitin brings a practical, end-to-end understanding of cell therapy development and commercialization.