Inclined sedimentation for selective retention of viable hybridomas in a continuous suspension bioreactor.
The continuous separation of nonviable hybridoma cells from viable hybridoma cells by using a narrow rectangular channel that is inclined from the vertical has been investigated experimentally. The effectiveness of the settler in selectively retaining viable hybridomas in the bioreactor while permitting the removal of nonviable hybridomas has been shown to depend on the flow rate through the settler. Intermediate flow rates through the settler have been found to provide the highest removal of nonviable hybridomas relative to viable hybridoma retention. At high dilution rates through the chemostat, over 95% of the viable cells could be partitioned to the bottom of the settler while over 50% of the nonviable cells are removed through the top of the settler. This successful separation is due to the significantly larger size of the viable hybridomas than the nonviable ones. A continuous perfusion experiment was performed in which an external inclined settler was used to retain virtually all of the viable hybridomas in the culture, while selectively removing from the culture approximately 20% of the nonviable cells that entered the settler. A stable viable cell concentration of 1.0 x 10(7) cells/mL was achieved, as was an antibody productivity of over 50 micrograms/(mL.day). These represent 3- and 6-fold increases, respectively, over the values obtained from a chemostat culture without cell retention.
Department of Chemical Engineering, University of Colorado, Boulder 80309-0424.
B C
BC
Batt
R H
RH
Davis
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DS
Kompala
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D002452:Cell Count; D002460:Cell Line; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D005434:Flow Cytometry; D006825:Hybridomas / Q000166:cytology*; D051379:Mice; D010477:Perfusion
D000911:Antibodies, Monoclonal