Scaling up from R&D to commercial production is a pivotal phase of the biopharmaceutical development process. Efficiency during this stage is critical to enable you to meet your planned commercialization timelines; however, this must be achieved without compromising on process performance or productivity.

Central to developing a scalable process and establishing an optimized scale-up strategy is a comprehensive understanding of your process. This should include knowledge of your cell line’s behavior and nutritional requirements, as well as your cell culture medium formulation and its performance. These insights are particularly important to help ensure product titer and critical quality attributes (CQAs) are consistently met at all scales. The logistics of large-scale manufacturing should also be evaluated ahead of scale-up to enable you to maintain process productivity as production volumes increase.

In this article, we outline the tools and techniques that can be leveraged to help you gain this process understanding and the key logistical considerations you should make ahead of scale-up. Additionally, we discuss the benefits that working with an experienced and knowledgeable external collaborator can bring, from offering additional analytical capabilities to providing in-depth process insights.

Navigating scale-up challenges with cell line understanding

As all biopharmaceutical manufacturing workflows are unique, there is no one-size-fits-all approach to scale-up. As a result, it is important to understand the areas where challenges can arise and thoroughly consider the specific requirements of your process to develop a tailored strategy.

One key challenge is the inherent variability of different cell lines, since even stable clones generated from the same parent cell line and vector system can often display clone-specific behaviors. These include differences in growth patterns and peak cell densities, which give rise to different optimal feeding strategies and overall nutrient requirements. There are also more subtle differences in cell line behavior that can be equally challenging to manage. For example, some cell lines can demonstrate less resilience to sparged gas-generated shear, or different quality profiles for produced proteins. As such, broad optimization strategies based on cell type alone may not lead to optimal results, making it critical to understand your cell line’s specific media and process requirements.

There are several analytical techniques that can help you gain a greater insight into your cell line’s nutritional requirements and identify the media components that are driving productivity. By helping you enhance key process parameters, including cell growth and viability, an optimal media formulation can help you more consistently meet your product titer targets and CQAs at all scales.

Cell culture analysis has been greatly enhanced over recent years with the development of new analytical techniques and approaches. Chromatographic techniques, namely high-performance liquid chromatography (HPLC) and liquid chromatography–mass spectrometry (LC–MS), have evolved significantly. This has helped increase both sensitivity (for quantitation of components with low concentration) and selectivity (to identify a greater number of components). These improvements have paved the way for multi-omics analysis.

Multi-omics combines the insights of proteomics, the study of proteins expressed in the cell, with metabolomics, the study of metabolites produced through intracellular metabolic pathways. Utilizing multi-omics can be highly beneficial, as it can help support a greater understanding of cell phenotype and the design of optimized media and feed formulations.

Understanding your formulation through media analytics 

Once suitable media and feed formulations have been developed, it is crucial to understand how they will perform at larger scales. To help reduce the risk of costly and time-consuming reformulation, you should consider validating the scalability and manufacturability of your formulation ahead of scale-up. One way you can do this is by working with a prototyping service to rapidly manufacture small batches of media that can be used to assess how the formulation will perform at commercial scales.

When choosing a prototyping service, you should prioritize looking for a vendor that harmonizes its process controls and quality management standards between its non-CGMP and CGMP manufacturing sites. Choosing a vendor that also utilizes CGMP-grade raw materials can be particularly beneficial. This can help maximize equivalency between prototype media and the final CGMP-manufactured product, supporting more accurate evaluation, simplifying technology transfer, and helping reduce the risk of scale-up delays.

More comprehensive analytical testing of your formulation can also help you implement robust quality control measures to reduce the risk of variability during commercial production. For instance, trace element testing can enable you to understand which trace elements your cell line or process is particularly sensitive to. This insight can be used to develop a comprehensive, routine raw material testing and monitoring program to help ensure consistency and optimal performance.

Cell culture media stability studies can also provide several advantages. By establishing the shelf life of your formulation in its final packaging, this analysis can help you set accurate expiration dates. This understanding can be invaluable to help you optimize media storage and enable you to consistently achieve reliable and reproducible performance.

Logistical considerations and choosing the right media format

Insufficient planning for the logistics of large-scale manufacturing during process development can also complicate scale-up and potentially lead to delays. Without careful consideration of your operational logistics, you could encounter shipping and storage challenges surrounding the transportation and management of higher volumes of bioprocessing components such as cell culture media and feeds. Liquid concentrates, formulations with higher nutrient density, and dry media formats have all been key in helping reduce the logistical challenges associated with managing the volume of media required for commercial-scale production.

Depending on your process, you may struggle to maintain process efficiencies as you transition toward clinical and commercial production. For instance, certain manual process steps like media preparation can take longer due to lower mixing efficiency at larger volumes. This is where advanced granular, dry media formats have had an even more transformative impact. By integrating the ease-of-preparation benefits of liquid media with the reduced shipping and storage requirements of dry media, granular formats can provide several operational advantages. Properties including pH and osmolality are pre-adjusted and the ability to develop complete, single-part formulations results in fewer manual interventions and can help improve consistency. In addition, the faster dissolution rates of granules compared to powders can increase efficiency, which becomes even more valuable at larger scales.

Improving your process understanding through collaboration

The range of tools and techniques that can be utilized to help you understand your cell line and media is extensive. Consequently, in-house capacity constraints, limited access to specialized equipment, or a lack of experience can make it challenging to implement them into your development workflow. Interpreting complex analytical data and converting the outputs into actionable process optimizations can be especially difficult without specific expertise. 

As a result, to support seamless scale-up, it can be beneficial to work with a knowledgeable collaborator with the analytical capabilities and expertise to help you gain a thorough process understanding. Using their resources, a collaborator can help you gain a deeper insight into your cell line and medium, as well as highlight any potential scale-up challenges that may arise. Moreover, they can help you manage your process logistics, providing support with format conversion and optimization of your supply chains.

Ultimately, with the guidance of an experienced collaborator, you can take proactive steps to minimize the risk of scale-up challenges, helping streamline your transition to sustainable large-scale production and long-term commercial success.

Find out how we can help you optimize scale-up with dedicated analytical, media development, and rapid prototyping support services at thermofisher.com/mediabydesign.

About the Author

Femi Egbebi, Senior Global Product Manager for Media Development Services, Bioproduction at Thermo Fisher Scientific.

Femi Egbebi is the Senior Global Product Manager for Media Development Services, Bioproduction, at Thermo Fisher Scientific. He has an MBA from Carnegie Mellon University, a PhD in Chemical Engineering from the Louisiana State University, and focuses his work on custom products and services for the bioprocessing industry.