Spin Filter Perfusion System for High Density Cell Culture: Production of Recombinant Urinary Type Plasminogen Activator in CHO Cells

• Published in: Bio/technology (New York, N.Y. 1983) Vol. 8; no. 1; pp. 54 - 58
• Main Authors: Avgerinos, George C, Drapeau, Denis, Socolow, Jeff S, Mao, Jen-i, Hsiao, Kathy, Broeze, Robert J
• Format: Journal Article
• Language: English
• Published: New York, NY Nature Publications 01-01-1990
• Subjects: Animals; Biological and medical sciences; Biotechnology; Biotechnology - methods; Cell Count; Cell Division; Cell Line; Cell Survival; Cloning, Molecular; Cricetinae; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; Gene Expression; Health. Pharmaceutical industry; Industrial applications and implications. Economical aspects; Other active biomolecules; Plasminogen Activators - biosynthesis; Plasminogen Activators - genetics; Plasminogen Activators - isolation & purification; Production of active biomolecules; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Transfection - genetics; Urokinase-Type Plasminogen Activator - biosynthesis; Urokinase-Type Plasminogen Activator - genetics; Urokinase-Type Plasminogen Activator - isolation & purification; Cell culture; Enzyme; Plasminogen activator; Mixed tank reactor; CHO cell line; Optimization; Cell density; Continuous process; Bioreactor; Production; Recombinant protein; Recycling; Viability
• ISSN: 0733-222X; 2331-3684
• DOI: 10.1038/nbt0190-54

We have used a 20 liter stirred tank fermentor, equipped with a 127 mesh ethylene-tetrafluoroethylene rotating screen for cell recycle, for the continuous production of recombinant single chain urokinase-type plasminogen activator (rscu-PA) from Chinese hamster ovary (CHO) cells. Viable cell densities between 60 and 74 million per ml were maintained at medium perfusion rates of 3.0 to 4.0 fermentor volumes per day. Cells were retained by the 120 micron nominal opening filter through the formation of "clumped" cell aggregates of 200 to 600 microns in size, which did not foul the filter. In 31 days of culture, a total of 51 grams of rscu-PA were produced in 1,000 liters of medium. The rscu-PA produced over the course of this continuous culture was purified and characterized both in vitro and in vivo and shown to be comparable to natural scu-PA produced from the transformed human kidney cell line, TCL-598.