PROTEOLYTIC ACTIVITY OF HYBRIDOMA CULTURES -A SIMULATION STUDY

摘要

Using a structured model written in the PC program "MATLAB", we have studied the influence of proteolytic activity on antibody concentration in hybridoma cell cultures and the effect of addition of a sacrificial substrate, which the proteinase could degrade instead of the antibody. These simulations were carried out in a factorial experimental design including the variation of the individual rate constants for breakdown kinetics between different cell lines. The results show that the differences in these rate constants are very important for the increase in antibody yield when a sacrificial substrate is added and that proteolytic activity can be present, even though it is not revealed by the antibody curve, indicating that product degradation in cell cultures may be more wide spread than normally assumed. Apparent variations in productivity of antibody may be related to the productivity of proteolytic enzymes and the modification of the activity of these enzymes by medium components, rather than intrinsic changes in cell behavior.