Rapid intensification of an established CHO cell fed-batch process

Area of Science:

• Biotechnology
• Bioprocess Engineering
• Mammalian Cell Culture

Background:

• Biomanufacturing requires process intensification (PI) for economic and flexible production of biotherapeutics.
• Conventional fed-batches (FBs) are slow to implement intensified processes due to extended development times.
• Rapid, efficient, and straightforward PI strategies are crucial for the biopharmaceutical industry.

Purpose of the Study:

• To develop and demonstrate a rapid PI strategy for N-stage fed-batch (FB) processes.
• To combine N-1 perfusion and high inoculum densities for an intensified FB (iFB).
• To evaluate the impact of PI on space-time yield (STY), product quality, and cellular metabolism, including the use of productivity enhancers.

Main Methods:

• Step-wise intensification of a conventional Chinese Hamster Ovary (CHO) cell FB process in parallel small-scale bioreactors.
• Implementation of N-1 perfusion and high inoculum cell densities to create intensified FBs (iFBs).
• Evaluation of key process indicators (KPIs) including space-time yield (STY) and product quality.
• Assessment of metabolic changes using flux balance analysis (FBA).

Main Results:

• Successfully doubled space-time yield (STY) from 0.28 to 0.55 g/L/d.
• Maintained product quality in the intensified fed-batch (iFB) process.
• Achieved enhanced cell-specific productivity from ~25 to 37 pg/(cell·d) by reducing cell growth with butyric acid (BA).
• Observed transient metabolic differences that resolved quickly, indicating process adaptability.

Conclusions:

• A robust and rapid PI strategy for mammalian cell culture biomanufacturing was demonstrated.
• The intensified fed-batch (iFB) process, combining N-1 perfusion and high inoculum densities, significantly improves space-time yield without compromising product quality.
• This approach allows rapid implementation for new and existing processes, showing minimal sustained impact on cellular metabolism.