Rapid development of clone-specific, high-performing perfusion media from established feed supplements.

Area of Science:

• Biotechnology and Biopharmaceutical Manufacturing
• Cell Culture Technology
• Process Optimization

Background:

• Perfusion cultivation of recombinant Chinese hamster ovary (CHO) cells is crucial for the biopharmaceutical industry due to enhanced space-time-yields (STYs) and reduced residence times.
• Economical bioprocesses require cultivation media that support rapid cell growth and high protein production with minimal consumption.
• Developing tailored media is essential for maximizing the efficiency of perfusion cultures.

Purpose of the Study:

• To present a straightforward and rapid two-step approach for developing clone-specific, high-performing perfusion media for recombinant CHO cells.
• To optimize media formulations by combining commercially available basal media and feed supplements using design-of-experiment (DoE).
• To achieve high cell densities and STYs in perfusion cultures.

Main Methods:

• A two-step strategy involving initial selection of feed supplements in batch cultures.
• Optimization of feed supplement mixing ratios in small-scale semicontinuous perfusion cultures.
• Statistical response surface modeling to finalize media blends, followed by verification in perfusion bioreactors.

Main Results:

• Two novel media blends were developed, enabling perfusion bioreactor runs to reach 200 × 10^6 cells/ml within two weeks.
• Achieved minimum cell-specific perfusion rates as low as 10-30 pL/cell/day.
• Obtained STYs ranging from 0.4-1.2 g/L/day, representing a 10-fold increase compared to batch cultures.

Conclusions:

• The presented two-step workflow is effective for developing high-performing perfusion media for recombinant CHO cells.
• The optimized media blends significantly enhance cell density and productivity in perfusion cultures.
• This general workflow is broadly applicable to various perfusion platforms, cell lines, and media components.