Analysis of nutritional factors and physical conditions affecting growth and monoclonal antibody production of the hybridoma KB-26.5 cell line

The effect of medium composition and physical conditions on the growth pattern and monoclonal antibody production of the hybridoma cell line KB-26.5 has been studied in batch and fed-batch cultures. Different aspects have been analyzed both individually and in combination, as it is expected that not only one component plays a predominant role in this process but a combination of a number of them. Modification of the concentrations of glucose and glutamine, supplementation of the medium with vitamins and amino acids, influence of lactate and ammonium accumulation, and pulse addition of fetal calf serum have been studied in detail, contributing to an improvement in the cell growth and IgG3 production. Finally, the combined effect of all these factors in a fed-batch culture leads clearly to a major improvement in the cultures, with a 10-fold increase in the final monoclonal antibody concentration with respect to nonoptimized batch experiments.