Implementing an Intensified Dynamic Perfusion Process for Biologics Manufacturing at Scale and Continued Development

ABSTRACT

Perfusion cell culture is increasingly adopted to improve productivity, reduce footprint, and enable integrated continuous bioprocessing. The high‐intensity, low‐volume perfusion (HILVOP) process for antibody production was implemented at both 100 L pilot and 500 L manufacturing scales for early clinical supply, and development work was completed in tandem to create a next‐generation HILVOP platform. The HILVOP process employs a dynamic two‑stage strategy, using cell‑controlled perfusion during the growth phase followed by product removal during the intensified production phase via tangential flow filtration (TFF). Scale‑up performance was evaluated across multiple CHO cell lines with a focus on membrane robustness, product recovery, and space‑time yield. Early manufacturing runs identified membrane fouling as a key risk, particularly with challenging cell lines. Systematic optimization of TFF operating conditions (reduced membrane wall shear rate, larger pore size, alternative filter membrane, and refinement of perfusion rate profiles) improved membrane performance, delayed transmembrane pressure (TMP) rise, and increased cumulative product recovery while reducing media consumption. Additional productivity gains were achieved through high‑density seeding and extension of the productive perfusion phase, resulting in increased permeate space‑time yield. These results demonstrate the robustness and scalability of the HILVOP platform and highlight key design considerations enabling reliable manufacturing‑scale perfusion operation.