High density cultivation of hybridoma in charged porous carriers

Abstract

A porous carrier whose surface is charged with polyethyleneimine (PEI) was evaluated for high-density cultivation of immobilized hybridoma cells. The carrier was used either in suspension or in fixed bed. For suspended carriers, the effect of carrier pore size, ion-exchange capacity (IEC), mixing intensity and inoculum concentration on the attachment and growth of HBs MAb-producing hybridoma cells in carrier CELLSNOW EX were studied. The carrier with 100 μm pore size was more efficient than that with 500 μm in supporting cell attachment. In addition, the amount of antibody which was detected by PHA (passive hemagglutination assay) was 4-times higher in 100 μm compared with 500 μm pore size. At low initial cell concentration, 2.8 × 10 5 ml −1, the attached cells did not grow in the first 7 d as indicated by a negligible glucose consumption. However, cell proliferation was observed at high inoculum, 9 × 10 5 ml −1. Hybridoma cells were expected to attach faster to EX with high IEC than with low IEC but results showed no significant difference. A different hybridoma cell line producing monoclonal antibody (MAb) against penicillin-G-amidase was successfully cultivated in an axial-flow fixed bed containing CELLSNOW carriers. The specific glucose uptake rate as an indicator for cell activity was 24-times higher compared with conventional chemostat cultures. The CELLSNOW carrier offers many advantages over other carriers.