Repeated fed-batch culture of hybridoma cells in nutrient-fortified high-density medium.

Long-term high-density cultivation of the hybridoma 2c3.1 was successfully carried out in a repeated fed-batch mode using high-density media that were constructed to meet in vitro cell growth limitations. The high-density culture was possible in a range of 0.5 approximately 1.0 x 10(7) cells/mL in MBRI 40-02 medium for over 2500 h by the repeated supplementation of the most fortified medium, MBRI 40-03, and consequently, distinct enhancement of MAb production was achieved. MAb concentrations were maintained around 1 g/L for about 1000 h of the process and the maximum MAb concentration was around 1.56 g/L. The result supported strongly the fact that the nutritional fortification was the most critical factor for high-density cell culture in vitro. The mean chromosome number of the hybridoma 2c3.1 was maintained stably for about 1500 h, whereas gradual loss of the MAb activity was apparent during the long-term cultivation. (c) 1993 John Wiley & Sons, Inc.