Perfusion Cell Culture Decreases Process and Product Heterogeneity in a Head-to-Head Comparison With Fed-Batch

Area of Science:

• Biotechnology
• Bioprocess Engineering
• Protein Therapeutics

Background:

• Fed-batch and perfusion are common cell culture platforms for monoclonal antibody (mAb) production.
• Optimizing these platforms is crucial for efficient and consistent therapeutic protein manufacturing.

Purpose of the Study:

• To compare the process and product attribute impacts of fed-batch versus perfusion platforms for IgG1 and IgG4 producing cell lines.
• To evaluate product quality attributes including purity, heterogeneity, and glycosylation under different culture conditions.

Main Methods:

• Bench-scale "plug-and-play" approach using commercial media and standard feed/filtration strategies.
• Process monitoring via Principal Component Analysis (PCA) and Lactate Dehydrogenase (LDH) assays.
• Product attribute analysis including charge variants, half-antibody formation, purity, and glycosylation.

Main Results:

• Perfusion achieved 7.5x higher average productivity than fed-batch, despite lower product concentration.
• Fed-batch cultures exhibited greater cell environment and metabolic variability, with 7-10x higher product exposure to cell lysate.
• Perfusion resulted in higher product purity, lower half-antibody formation, and increased abundance of neutral species, indicating reduced intra-lot heterogeneity.

Conclusions:

• Perfusion cultures demonstrate decreased intra-lot heterogeneity for monoclonal antibody production compared to fed-batch.
• While glycosylation remained similar, initial perfusion harvests showed distinct profiles, suggesting a lag between product quality and metabolic state.
• Further investigation is needed to determine the developmental, clinical, and commercial implications of enhanced uniformity offered by perfusion.