ObjectivesThe 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).ResultsThe 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 degrees C and a circulation rate of 182.5mL/min could support the hybridoma had the maximal antibody titer of 60.75 mu g/mL for 20days. When the ammonium ion was 65ppm or lactate close to 2.6g/L, the medium was replaced to maintain the stable and healthy cells at the high cell concentration of 3.33x10(8)/mL for continuous antibody production. Two serum-free media could be successfully applied to this perfusion system and maintain hybridoma growth and antibody production.ConclusionThe single-use HFBs could provide the advantages including high cell density, low shear stress, and continuous antibody production.