Continuous stable production of von Willebrand Factor monoclonal antibody in spin filter bioreactor with bleeding technology

• Published in: Biotechnology and bioprocess engineering 2000-04, Vol.5 (2), p.130-135
• Main Authors: Yun, Jeong-Won, Lee, Soo-Young, Choi, Byung-Wook, Oh, Han-Kyu, Kim, Se-Ho, Byun, Tae-Ho, Park, Song-Yong
• Format: Article
• Language: English
• Subjects: Ammonia; Aquaculture; Bioreactors; Bleeding; Cell culture; Cell density; Cell viability; Filters; Hybridoma; Kinetics; Metabolites; Microbiology; Monoclonal antibodies; Nutrients; Oxygen consumption; Perfusion; Von Willebrand factor
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/BF02931884

The characteristics of two different modes of perfusion culture, intermittent and continuous bleedings, were investigated by culturing the hybridoma cells producing von Willebrand Factor (vWF) monoclonal antibody (McAb) in a 15 L bioreactor without clogging the filter. Both culture methods exhibited similar profiles of cell density and metabolite concentrations during the culture period at the cell concentration of around 1×10^sup 7^ cells/mL. When the perfusion rate was increased, the intermittent bleeding culture showed problems of ammonia accumulation and decrease of cell viability. The continuous bleeding culture exhibited higher physiological activity than that of the intermittent bleeding culture in terms of nutrient consumption and metabolite production kinetics. But the analysis of specific oxygen consumption rate showed that the specific oxygen consumption rate of intermittent bleeding culture was similar to that of exponential growth phase. The continuous bleeding culture showed higher specific vWF McAb productivity and cumulative production than those of the intermittent bleeding culture. Finally we proved the possibility of long-term operation of continuous bleeding culture and produced approximately 40 g of vWF McAb in a 15 L bioreactor after one-month operation.[PUBLICATION ABSTRACT]