Enabling Cell Line Tech Transfer For mAb Platform Manufacturing

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By Steve Loftus
Business Steering Group Lead, FUJIFILM Diosynth Biotechnologies　

　 

The biopharmaceutical industry aims to deliver efficiently innovative therapies to those in need. Technology transfer, the process of transferring bioprocess knowledge, skills and technology, is a pivotal mechanism for ensuring adequate and timely access to medicines. Technology transfer occurs between a sending site and a receiving site, which can be within the same facility or across different geographies and even different companies.

Technology transfer supports clinical development teams by supplying material for pre-clinical, clinical and commercial efforts. The need for technology transfer can arise at any stage of the drug lifecycle. During early development phases, biotechs often use technology transfer to increase production capacity by moving manufacturing processes from a development laboratory to a facility where there is the expertise and experience to complete the full suite of CMC development, as well as the manufacturing processes required to support the entire product lifecycle (pre-clinical, clinical and commercial process development and manufacturing). Successful early-phase transitions are crucial for accelerated pivotal trial testing, enabling maximal patient benefit more quickly.

Substantial platform knowledge, product development history, cGMP experience and facilities have been established for mAbs produced by CHO cell lines, yielding a low-risk development and manufacturing profile. This enables a rapid development model based on platform expertise, manufacturing technology and infrastructure. However, it is commonly assumed that even low-risk mAbs require custom process optimization during technology transfer to meet the unique needs of a specific cell line, molecule or manufacturing facility. This perception arises from the belief that integrating an external cell line into a contract development and manufacturing organization’s (CDMO) standardized platform process is more complex and potentially less efficient than developing a custom process.

Advancements in technologies and expertise supporting the process development and manufacturing of mAbs has enabled greater flexibility in the biopharmaceutical industry. Many of today’s emerging biopharmaceutical companies are highly specialized, and the expertise of the right CDMO can be invaluable, particularly in streamlining and standardizing processes for larger production scales that support clinical and commercial manufacturing. Bridging the gap between discovery and manufacturing for products like mAbs often requires a balance between standardization and flexibility.

FUJIFILM Diosynth Biotechnologies (FDB) has been able to demonstrate the adaptability of its mAb platform process for externally developed cell lines using Sartorius’ 4Cell® CHO platform (Figure 1A). By performing comprehensive technology transfer in partnership with Sartorius, FDB was able to verify its established process, perform seed train expansion, and populate a production bioreactor, comparing culture parameters, growth, critical quality attributes (CQAs) and other key variables to validate a reproducible, scalable manufacturing process.

Case Study: Establishing A Successful Partnership

Figure 1A. An overview of the streamlined collaboration approach to linking Sartorius CLD services to FDB’s mAb Platform services.

Working in partnership with Sartorius, FDB was able to demonstrate the adaptability of its mAb platform process using a Sartorius cell line developed using their 4Cell® CHO platform (Figure 1B). Working with its partner, FDB performed a 4-week comprehensive technology transfer to understand Sartorius’s cell line and process in order to align with FDB’s mAb platform upstream process and to mobilize the cell line, reference materials, and consumables. FDB’s Ambr® 250 establishment run demonstrated comparability between FDB’s and Sartorius’s respective process performance. The 35-day FDB platform process’ robustness and scalability were successfully evaluated at 10 L scale, revealing comparable growth performance, product formation and critical quality attributes (PQA). This case study serves as evidence of a successful demonstration of FDB mAb Platform Process scalability using an external cell line.

Figure 1B. Workflow implemented to establish the partnership between Fujifilm and Sartorius.

The Process

The culture was progressed every three days through a 5-stage shake flask expansion and a 2-stage seed bioreactor expansion, using FDB’s platform conditions in order to provide enough material to inoculate the production vessels (either Ambr® 250 or 3x XDR 10) (Figure 2). During the Ambr® 250 run, FDB evaluated the Sartorius process (utilizing Sartorius’ 4Cell® SmartCHO media) and FDB platform process side-by-side, while during the 10 L run FDB evaluated different feeding strategies and scalability.

Figure 2. Process overview from vial thaw to production stage. FDB platform consists of a 5-stage shake flask expansion followed by 2 seed bioreactor before initiating the 14-day production phase, resulting in a 35-day process.

Growth Performance

Seed Train Expansion

Figure 3A shows comparable growth metrics between the Sartorius and FDB platforms during the shake flask expansion, demonstrating adaptability and robustness of the FDB platform, even when using non-platform media. The introduction of the seed bioreactors to align the Sartorius process to FBD platform was successful and demonstrated good facility fit and scalability (Figure 3B). On target transfer day (day 3), culture was above the target transfer for VCD, as per FDB platform.




Figure 3. Culture growth and viability profiles of (A) Shake flask expansion and (B) N-2 (Wave 25) and N-1 (XDR 10) seed expansion bioreactors.

Production Bioreactor

During the 14-day fed-batch production phase, growth, metabolites, gases and product formation were measured. Culture was fed from day 3 to day 13 with Sartorius feeds (4Cell® SmartCHO). During the production phase, growth metrics were shown to be comparable across scales, facilities and feeding strategies (Figure 4). The comparable growth metrics in the Ambr® 250 supports a straightforward technology transfer from Sartorius Process into FDB Platform Process.

The VCD and viability in the Ambr® 250 (FDB), 5 L (Sartorius) and 10 L (FDB) runs trend very close together, demonstrating the versatility, scalability and robustness of the FDB platform and Sartorius cell line across different production systems. The efforts to improve cell line manufacturability by simplifying the feeding strategy was successful as no detrimental effect is observed in growth characteristics between the three 10 L bioreactors.

Figure 4. Production bioreactors (A) Growth and (B) Viability profiles. Ambr® 250 data presented as mean ± 2SD (n=3).

Metabolites

A well-balanced feeding strategy is essential for high culture performance. Carbon limitation or high lactate levels can have a detrimental effect on culture performance and product quality attributes. Glucose levels were well controlled throughout the runs, across the different scales, demonstrating that the FDB glucose feeding strategy is scalable, robust and suitable to Sartorius cell lines. Lactate levels remained low throughout the runs, across the different scales, with peak lactate below 1.5 g/L and around day 3.

All other measured outputs, such as glutamine, glutamate, ammonium and osmolality were also comparable across bioreactors, scales and production systems (data now shown), once again demonstrating a straightforward technology transfer from Sartorius Process into FDB Platform Process, as well as the scalability of the FDB platform process.

Figure 5. Production bioreactors (A) Glucose and (B) Lactate profiles. Ambr® 250 data presented as mean ± 2SD (n=3).

Product titer and quality

Titer was measured from mid-run to day 14 (Figure 6). Profiles are comparable across bioreactors, scales and production systems, reinforcing the suitability of the FDB Platform to cultivate Sartorius cell lines. Product quality is also comparable across scales, indicating a well-controlled, scalable process.

Figure 6. Production bioreactors (A) Titer by Protein A UPLC and (B) Critical product quality attributes. Ambr® 250 data presented as mean ± 2SD (n=3).  

Scalability

The next step for this partnership would be to demonstrate FDB Platform Process scalability at pilot scale (200 L) while using the Sartorius cell line. Current FDB internal data already demonstrates the FDB platform process’ scalability potential. Using a well-defined and understood production process alongside standardized equipment, FDB is able to deliver an efficient, robust and scalable process that is seamlessly transferrable to manufacturing without requiring a time-consuming optimization phase.

Figures 7A, 7B, 7C and 7D below:

Figure 7. FDB mAb platform scalability across several projects. Production bioreactors (A) Growth, (B) Viability, (C) pCO2 and (D) Titer and productivity.

Beyond The Case Study: Routes To Manufacturing

The capabilities FDB brings to bear for its customers touch nearly every aspect of biologics and advanced therapeutic production, from early characterization and optimization to late-stage, large-scale commercial manufacturing. These capabilities are supported by a network of state-of-the-art manufacturing facilities located across the globe, each highly flexible and specialized in different manufacturing technologies and scales. They include:

Expansions to FDB’s manufacturing facilities in the U.S. and Europe are part of its Partners for Life strategy, which aims to build capacity in large-scale production to support biopharma customers’ end-to-end needs and provide supply chain resiliency. In combination with FDB’s efforts to create a platform process that can be leveraged in any of its facilities, this interconnected and expansive network of manufacturing capacity is available to customers anywhere in the world. Its Denmark and Holly Springs facilities each offer multiple 20,000 L stainless steel bioreactors; in its facilities in the UK and US FDB can offer single-use stirred tank bioreactor systems up to 2,000 L. Capacity expansion at the Billingham, UK, site will also see the completion of a new fully single use flexible cell culture facility offering bioreactor scale of up to  5,000 L, enabling a network of adaptable large-volume production at every scale.

Different Cell Line, Same Result: The Value of FUJIFILM Diosynth Biotechnologies’ mAb Platform

Though FDB can perform cell line development (CLD) for clients, its partnership with Sartorius enables access to additional CLD services, creating new shared capacity; moreover, by transferring Sartorius’ CHO line to its own platform, FDB has been able to demonstrate the platform’s ability to take on cell lines developed outside FDB. In addition, its partnership with Sartorius gives FDB clients access to CLD that can produce a range of complex molecules, including:

  • Afucosylated antibodies
  • Non-traditional molecules and difficult to express constructs (enzymes, cytokines, bispecifics, trispecifics, etc.)
  • 2nd generation cell lines or cell line re-development programs
  • Biosimilars

Conversely, Sartorius CLD clients now have access to a CDMO that can seamlessly transition their molecule to process development and manufacturing, minimizing risk and enabling streamlined scaleup. From early manufacturability assessments to commercial cGMP manufacturing, FDB has significant experience in mAbs, having produced more than 65 CHO cell lines for mAb expression and engaged in over 100 CHO programs and eight commercially approved mAb programs. Its offerings include:

  • CLD, including expression studies, manufacturability assessments, product characterization, and early material supply
  • Process Development, including its mAb platform, cell culture PD, formulation PD, stability studies, process optimization, characterization, and validation
  • cGMP Manufacturing, including cell banking, drug substance scale-out and scale-up, and fill/finish

While FDB is well-versed in conventional process development, its mAb platform, where applicable, can enable the rapid implementation of processes and analytical methods, and offer the shortest and most predictable timelines and costs. This high-yield platform, structured around single-use technologies (SUTs), boasts both standard and flexible workflows and rapid implementation, generating tox material as early as six months and drug product roughly one year from the start of CLD.

Conclusion

Successful technology transfer is paramount for the efficient and timely delivery of innovative biopharmaceuticals to patients. By bridging the gap between development and manufacturing, technology transfer enables optimal production scale-up, ensuring adequate supply to meet clinical and commercial demands. FDB’s case study in partnership with Sartorius demonstrates the potential of standardized, flexible approaches like FDB’s mAb platform process to accelerate development and reduce risk. Operators were able to demonstrate the versatility, scalability and robustness of the FDB platform with an externally developed CHO cell line across different production systems, and likewise, to show that its platform processes are positioned to scale processes like this into large-volume systems while retaining quality and productivity.

By fostering collaboration and knowledge sharing between biopharmaceutical companies and CDMOs, the industry can optimize technology transfer processes, ultimately improving patient outcomes. Ultimately, partnering with a CDMO equipped with both a robust platform process and established partnerships can significantly expedite technology transfer and streamline mAb production. Through its successful collaboration with Sartorius, FDB has showcased the adaptability of its mAb platform for an externally sourced CHO cell line. This partnership not only validates FDB’s established process, but also facilitates seamless tech transfer and scaleup, minimizing the need for lengthy optimization and offering companies a more comprehensive path to commercialization.