The use of continuous culture to enhance monoclonal antibody production.

Hybridoma 14-4-4S, ATCC HB32, was grown in batch and continuous culture in an attempt to increase the production of monoclonal antibody (MAb), and to determine the adequacy of each culturing method for optimizing culture media. Various concentrations of glucose, fetal bovine serum (FBS) and serum type were studied to determine their effects on the specific growth rate (mu), the specific monoclonal antibody production rate (MPR), the maximum population density and the final MAb concentration. Attempts to optimize the culture medium using the batch method resulted in ambiguous results. However, by growing the hybridoma in a cytostat at a standardized population density of 1.0 X 10(6) cells/ml, the growth rate and MPR were found to decrease below feed concentrations of glucose and FBS of 22.5 mM and 10% (v/v), respectively. In addition, the use of the cytostat demonstrated that FBS was superior to both new born bovine serum (NBS) and Serum PlusTM (SPS). Batch cultures indicated that the production of MAb appeared to be related to the metabolic activity of the cells, and continuous culture demonstrated a direct relationship between mu and MPR.