Enhancing productivity for animal health mAbs: from fed-batch to perfusion
Introduction
Enzene Biosciences partnered with a global leader in specialty veterinary pharmaceuticals seeking to improve the productivity of a felinized monoclonal antibody (mAb) program. The client had already developed the production clone at another CDMO and completed an initial fed-batch run, however, titers were low and the process showed limited scalability, considering the commercial viability of the program. The client approached Enzene to determine whether the molecule could be transitioned to perfusion using Enzene’s fully-connected continuous manufacturing™ (FCCM™) technology. Enzene ran both fed-batch and FCCM perfusion processes using the same clone, generating a side-by-side comparison of productivity and product quality. Early findings indicate that the FCCM perfusion process delivered significantly higher cumulative productivity, approximately five-fold, based on reported yields (9.1 g vs. 45 g, while also maintaining higher cell viability.
Why did the customer approach Enzene?
The client, developing a mAb for the treatment of feline companion animals, arrived with a clone generated at a different CDMO. Their initial fed-batch production run yielded low titers, constraining both cost efficiency and the development timeline. Low titers also raised concerns about downstream processing costs and the overall manufacturability of the molecule.
Key challenges included:
- Suboptimal fed‑batch performance:
The initial titers generated by the original process were too low to support a commercially viable manufacturing strategy, leaving the client exposed to higher cost of goods and limited scalability.
- Need for rapid evaluation of an alternative process mode:
The client wanted to understand whether their molecule was a good candidate for perfusion, particularly using Enzene’s FCCM platform, with the goal of lowering COGS. A fast turnaround was important to maintain development momentum.
- Maintaining product quality during process intensification:
Shifting from fed-batch to perfusion can improve volumetric productivity but may introduce differences in glycosylation, charge variants, or other CQAs. Although regulatory expectations differ for animal health candidates, product consistency remains essential, with the client defining strict CQA ranges: aggregates <4%, fragments <3%, acidic and basic charge variants <40%, and a glycosylation profile with >80% complex fucosylated species.
- Ensuring clone compatibility with continuous manufacturing:
Clones developed under traditional fed-batch assumptions do not always adapt seamlessly to perfusion. Enzene had to determine whether the existing clone would sustain long-term viability under intensified conditions.
This combination of low productivity, uncertainty about process adaptability, and the need for improved manufacturability positioned Enzene to evaluate the feasibility of a conversion to perfusion.
Enzene’s approach
Enzene applied its FCCM™ technology to evaluate the molecule under both fed‑batch and continuous conditions using the same clone. This allowed a clean, controlled comparison of the two process modes.
Process Evaluation Approach
Enzene executed two parallel development tracks:
- Fed‑batch run using Enzene’s platform process
This served as a baseline for direct comparison.
- Perfusion run using FCCM™
The FCCM™ system is designed for high-productivity perfusion with controlled media exchange and stable operating conditions. The perfusion run produced 45 g cumulatively, compared with 9.1 g from fed-batch.
Key observations included:
- Sustained high viability across an extended culture duration, with perfusion maintaining ~98–99% viability through the first 10 days and remaining above 90% even at day 25, demonstrating a far more stable culture compared to fed-batch
- Longer productive culture window, with perfusion supporting continuous production for 25 days versus ~12 days in fed-batch, enabling significantly greater cumulative output
- Higher peak viable cell densities, reaching >70 million cells/mL during the perfusion run, compared with ~21 million cells/mL in fed-batch, contributing to improved productivity
- More consistent cell–growth trajectory, with perfusion showing a smooth rise to peak VCD and a gradual decline, whereas fed-batch exhibited an earlier drop-off in both VCD and viability
- Steady accumulation of product over time, with perfusion achieving >48 g cumulative productivity by day 25 versus ~9 g in fed-batch by day 12, reflecting both higher daily productivity and longer culture duration.
Product Quality Comparison
Quality attributes remained consistent between fed-batch and perfusion, with no adverse shifts observed during the FCCM run. The high viability maintained throughout the extended perfusion culture supported a stable product profile, confirming that the intensified process preserved the molecule’s critical quality attributes while delivering substantially higher productivity.
Benefits of Perfusion Using FCCM™
- 5x increase in cumulative productivity, directly impacting cost efficiency and scalability
- Higher cell viability throughout the run, suggesting that the clone adapted well to continuous conditions
- Feasibility of converting a fed-batch program to perfusion demonstrating the robustness of Enzene’s FCCM platform for clones developed externally.
Conclusion
Enzene successfully demonstrated the feasibility and advantage of transitioning the client’s felinized mAb from a low-yield fed-batch process to a high-productivity FCCM™ perfusion process, achieving a roughly five-fold productivity increase with higher cell viability. This outcome suggests that the molecule is well-suited for continuous manufacturing and that Enzene’s platform can rapidly evaluate and optimize processes for clones developed outside its facility.
Find Out More
To learn how Enzene can support your animal health development program, including the benefits of using Enzene’s fully-connected continuous manufacturing™ (FCCM™) technology, etc., contact our scientific team or visit our resources page at www.enzene.com.
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