Vortex flow filtration of mammalian and insect cells.
The use of vortex flow filtration for harvesting cells or conditioned medium from large scale bioreactors has proven to be an efficient, low shear method of cell concentration and conditioned medium clarification. Several 8-10 L batches of the human histiocytic lymphoma U-937 cell line (ATCC CRL 1593) were concentrated to less than 1 L by vortex flow filtration through a 3.0 microns membrane. An aggressive filtration regimen caused a 17% loss of cell viability and a 32% loss of IL-4 receptor binding capacity when compared to a batch centrifuged control. A reduction of the rotor speed from 1500 to 500 RPM and reduction of system back pressure from 10 to 0 PSIG resulted in cell viability and IL-4 binding capacity comparable to the control. Several 10 L batches of baculovirus infected Sf-9 cells were also concentrated to less than 1 L by vortex flow filtration through a 3.0 microns membrane. SDS-PAGE analysis of filtrate samples showed that aggressive filtration caused cell damage which led to contamination of the process stream by cellular lysate. When rotor speed was reduced to 500 RPM and system back pressure was reduced to 0 PSIG, the amount of contaminating lysate proteins in filtrate samples was comparable to a batch centrifuged control.
Hawrylik
S J
SJ
Pfizer Central Research, Groton, CT 06340.
Wasilko
D J
DJ
Pillar
J S
JS
Cheng
J B
JB
Lee
S E
SE
D000818:Animals; D001709:Biotechnology / Q000295:instrumentation / Q000379:methods; D002460:Cell Line; D002470:Cell Survival; D017077:Culture Media, Conditioned; D046508:Culture Techniques / Q000295:instrumentation* / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D005374:Filtration / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D016403:Lymphoma, Large B-Cell, Diffuse; D008322:Mammals; D008565:Membrane Proteins / Q000302:isolation & purification; D019948:Receptors, Interleukin-4; D011975:Receptors, Mitogen / Q000032:analysis / Q000096:biosynthesis; D018411:Spodoptera; D014407:Tumor Cells, Cultured
D017077:Culture Media, Conditioned; D008565:Membrane Proteins; D019948:Receptors, Interleukin-4; D011975:Receptors, Mitogen