A perfusion culture system using a stirred ceramic membrane reactor for hyperproduction of IgG2a monoclonal antibody by hybridoma cells

作者：Dong Haodi; Tang Ya Jie; Ohashi Ryo; Hamel Jean Franç ois P^*

来源：Biotechnology Progress, 2005, 21(1): 140-147.

DOI：10.1021/bp049826l

• Cell densities
• Cell growth-limited phase
• Cell retention devices
• Nutrient-limited condition

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 (ΔD, vvd per day, vvdd) of perfusion rate, the maximal viable cell density of 3.5 × 107 cells/mL was obtained within 6 days without any limitation and the cell viability was maintained above 95%. At lower ΔD's, the cell growth became limited. Under nutrient-limited condition, the specific cell growth rate (μ) was regulated by ΔD During the nonlimited growth phase, the specific mAb production rate (qmAb) remained constant at 0.26 ± 0.02 pg/cell&middoth in all runs. During the cell growth-limited phase, qmAb was regulated by AD within the range of 0.25-0.65 vvdd. Under optimal conditions, q mAb of 0.80 and 2.15 pg/cell&middoth was obtained during the growth-limited phase and stationary phase, respectively. The overall productivity and yield were 690 mg/L&middotday and 340 mg/L&middotmedium, 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.

• 出版日期2005