Multivariate analysis of metabolic parameters and optimization of antibody production using high cell density hybridoma in hollow fiber bioreactors

Objectives: The relationships of manipulation of culture temperature and medium circulation rate on the metabolic parameters were regressed by multiple linear regression analysis in hollow fiber bioreactors (HFB). Results: The high circulation rate could significantly enhance the oxygen consumption of the hybridoma cells and the medium’s oxidation–reduction potential. A mildly hypothermic condition of 36 °C and a circulation rate of 182.5 mL/min could support the hybridoma had the maximal antibody titer of 60.75 μg/mL for 20 days. When the ammonium ion was 65 ppm or lactate close to 2.6 g/L, the medium was replaced to maintain the stable and healthy cells at the high cell concentration of 3.33 × 10⁸/mL for continuous antibody production. Two serum-free media could be successfully applied to this perfusion system and maintain hybridoma growth and antibody production. Conclusion: The single-use HFBs could provide the advantages including high cell density, low shear stress, and continuous antibody production.