A Perfusion Culture System Using a Stirred Ceramic Membrane Reactor for Hyperproduction of IgG_(2a) Monoclonal Antibody by Hybridoma Cells

摘要

A novel perfusion culture system for efficient production of IgG_(2a) monoclonal antibody (mAb) by hybridoma cells was developed.A ceramic membrane module was constructed and used as a cell retention device installed in a conventional stirred-tank reactor during the perfusion culture.Furthermore,the significance of the control strategy of perfusion rate (volume of fresh medium/working volume of reactor/day,vvd) was investigated.With the highest increasing rate (AD,wd per day,vvdd) of perfusion rate,the maximal viable cell density of 3.5 x 10~7 cells/mL was obtained within 6 days without any limitation and the cell viability was maintained above 95%.At lower AD's,the cell growth became limited.Under nutrient-limited condition,the specific cell growth rate (mu) was regulated by AD.During the nonlimited growth phase,the specific mAb production rate (q_(mAb)) remained constant at 0.26+-0.02 pg/cell-h in all runs.During the cell growth-limited phase,q_(mAb) was regulated by AD within the range of 0.25-0.65 wdd.Under optimal conditions,q_(mAb) of 0.80 and 2.15 pg/cellcentre doth was obtained during the growth-limited phase and stationary phase,respectively.The overall productivity and yield were 690 mg/L-day and 340 mg/L-medium,respectively.This study demonstrated that this novel perfusion culture system for suspension mammalian cells can support high cell density and efficient mAb production and that AD is an important control parameter to regulate and achieve high mAb production.