50L Scale Perfusion Culture of Hybridoma Cells by Gravitational Settling for Cell Separation

Summary

A mouse - human hybridoma, X32 was cultured in a perfusion system with a gravitational settling zone for cell separation. The viable cell density and IgG concentration reached 10⁷ cells/mL and 30 mg/L respectively in 50L scale, which were almost equivalent to those in the smaller culture.

In the case of scaling-up a gravitational settling type of perfusion culture vessel, the larger settling area is required for cell separation in proportion to the culture volume. As the culture volume increases, the cell separation becomes more difficult because it is susceptible to be influenced by any liquid turbulence in the settling zone.

Then, the clear separation of cells from the culture medium was achieved by improving agitation and controlling liquid turbulence in the settling zone. In consequence, we succeeded in 4L, 20L and 50L perfusion culture as in 120 mL one.

The system reported here satisfied the requirement in practical application.