High-level production of a monoclonal antibody in murine myeloma cells by perfusion culture using a gravity settler.
A perfusion system is described for the production of a human monoclonal antibody in non-secreting murine myeloma (NS0) cells that was previously shown to be difficult to produce at high levels using fed-batch culture. The perfusion system was based on the use of a commercially available cell settler as the separation device to separate the cells from the culture. Separation efficiency of the cell settler was above 98%. Based on the growth and glucose consumption rates, fresh media was added to the culture and the turnover rate for the bioreactor was set at a maximum of 1.5 times the bioreactor volume per day. The perfusion process resulted in twice the maximum viable cell densities and up to three times the total protein production in a 53-day run period when compared to the fed-batch process. In addition, charge heterogeneity of the antibody as measured by ion exchange chromatography was lower for material purified from the perfusion runs compared to fed-batch. Perfusion mode of culture using a commercially available gravity settler is therefore a viable alternative to fed-batch mode for high-level production of this monoclonal antibody in NS0 cells.
Choo
Chiou-Yu
CY
Department of Process Development, Human Genome Sciences Inc., 14200 Shady Grove Road, Rockville, Maryland 20850, USA.
Tian
Yuan
Y
Kim
Wan-Seop
WS
Blatter
Erich
E
Conary
Jon
J
Brady
Ciaran P
CP
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification* / Q000378:metabolism*; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D002469:Cell Separation / Q000379:methods*; D002498:Centrifugation / Q000379:methods*; D051379:Mice; D009101:Multiple Myeloma / Q000378:metabolism*; D010477:Perfusion / Q000379:methods*
D000911:Antibodies, Monoclonal