In this application we substituted an optional NBS spin filter with a marine blade impeller for the pitched blade impeller, (Fig. 2). The spin filter impeller is very useful for a continuous perfusion process because it is a simple device that lends itself to media exchange and can be easily scaled up. (See Appendix.)
With most commonly-available spin filters, screens rapidly become clogged when cells reach high cell density during perfusion. Therefore, NBS designed spin filters with three different screens for various cell sizes. Please contact us for screen size suggestions.
All six control modules included in the Advanced Cell Culture Kit were used for this run:
Materials and Methods
HyQ SFX-Insect (liquid) medium supplied by Hyclone (Logan, UT) (serum-free expression medium).
Hi-5 insect cell inoculum supplied by Hyclone (Logan, UT) Inoculum was cultivated in a NBS Shaker (Innova 2000) which was inside an Incubator (Fisher Scientific, Model No. 10).
Sterilize the vessel with phosphate buffer solution (PBS) for 60 minutes.
Remove the PBS from the vessel.
Add 1.5 L of insect cell media to the vessel.
Inoculate 500 ml of insect cell suspension to provide a starting cell count of 4.1 x 105 cells/ml.
Setpoints were keyed into the controller prior to inoculation. The vessel was allowed to equilibrate prior to inoculation. DO loops current value rem