Cellicon
TM
Perfusion Solution for Process Development User Guide
18
Operating Parameters
The bench scale N-1 perfusion filter was
designed for N-1 perfusion processes
targeting:
• 3-5 days of perfusion
• 2.7 L working volume
• 2 VVD
• 50 million cells/mL cell density
The performance of the filter is specific to
the cell line and media combination, as well
as the chosen operating parameters. The
most critical factors that impact performance
are perfusate (permeate) flux, which should
be less than 23 L/m
2
/hr, cell density, and
crossflow rate.
Starting a Run
Priming
The recirculation pump for the cell retention
device is a magnetically coupled centrifugal
pump. It enables the assembly to be
single use and disposable while providing a
consistent and low shear flow rate. The pump
does not self-prime (will not successfully
pump unless it is filled with liquid).
When the bioreactor is in media hold and
the cell retention filter has been attached,
a priming procedure is performed to start
recirculation. Due to the harvest port location
on the Mobius
T
M
3L, it may prime after
unclamping the feed line. In this case, the
tubing will fill with liquid and recirculation
flow can start from the controller. If the
tubing does not self-prime, a secondary
peristaltic pump is required to start the
flow recirculation. Step-by-step priming
instructions can be found by clicking
Prime
system
on the main menu screen.
Priming Procedure
1. Select
Prime system
on the main menu
screen. This will switch the controller into
Speed setpoint
mode and provide step-
by-step guidance on how to complete the
priming.
2. Ensure that the perfusate (permeate) line
is clamped.
3. Take an external peristaltic pump and
place the pump head on the feed line of
the cell retention device loop.
4. Turn on external peristaltic pump (<100
mL/min) and allow liquid to flow until
there are no bubbles left in the feed line
or pump head. If necissary, adjust the
angle to remove all bubbles.
5. Stop external peristaltic pump but leave it
clamped so bubbles do not return to the
feed line.
6. Select
START PUMP
on the controller
with the speed setpoint at 3000 RPM.
Quickly remove the external peristaltic
pump from the feed line, allowing flow to
recirculate through the device. Increase
the RPM to clear bubbles from the loop.
7. Select
Exit Priming
(returns to flow
setpoint mode). The flow will adjust
to the setpoint that was previously
entered in flow setpoint mode. Adjust
the flow setpoint according to the desired
recirculation rate for the run (100 mL/
min).
8. Select
Zero adjust
from the control
region of the main screen. Refer to
Turning the Pump On
The system is ready to begin recirculation
flow once the flow path has been primed and
the sensors have been zero adjusted.
There are two ways to operate the pump
between
Flow setpoint
and
Speed setpoint
,
indicated by the toggle on the main screen.
Ensure that
Flow setpoint
is selected for the
perfusion run. Speed setpoint is intended
to be used for priming only. See below for a
description of each setpoint:
