REPLIGEN KrosFlo TFDF User manual

1
IF.UG.023 R1
KrosFlo® TFDF™
Lab System
User Guide

KrosFlo® TFDF™ Lab System
User Guide
KrosFlo® TFDF™ Lab System
User Guide
2
IF.UG.023 R1
The information contained in this document is subject to change without notice.
Repligen Corporation makes no warranty of any kind with regard to this material, including, but not
limited to, the implied warranties of merchantability and fitness for a particular purpose.
Repligen Corporation shall not be liable for errors contained herein or for incidental or
consequential damages in connection with the furnishing, performance, or use of this material.
No part of this document may be photocopied, reproduced, or translated to another language
without the prior written consent of Repligen Corporation.
For further information, please contact Repligen Corporation at www.repligen.com.
©2020 Repligen Corporation. All rights reserved. The trademarks mentioned herein are the property
of Repligen Corporation and/or its affiliate(s) or their respective owners.
Customer Support
508-845-6400
customerserv[email protected]om
Repligen Corporation
111 Locke Drive
Marlborough, MA 01752
www.repligen.com

KrosFlo® TFDF™ Lab System
User Guide
KrosFlo® TFDF™ Lab System
User Guide
3
IF.UG.023 R1
Contents
Optional connections..................................................................................................16
20 cm flow path installation .......................................................................................18
108 cm flow path installation .....................................................................................19
Flow path pressure sensor connections .....................................................................21
Tubing connections.....................................................................................................22
Pump priming set-up ..................................................................................................26
Pump priming process ................................................................................................27
Changing the default tubing calibration factors .........................................................35
Changing the system serial number ...........................................................................35
Updating the permeate flow meter installation status..............................................36
Setting tubing sizes .....................................................................................................36
Scaling .........................................................................................................................37
Changing the tubing calibration factors for a run (not default) .................................37
Resetting Tubing Calibration factors...........................................................................37
Setting the maximum magnetic levitating recirculation/feed pump (P-01) speed....38
Setting pressure units .................................................................................................38
Setting the Permeate Side Hold-up Volume...............................................................38
Alarm Set-up screen....................................................................................................40
Alarm History ..............................................................................................................42

KrosFlo® TFDF™ Lab System
User Guide
KrosFlo® TFDF™ Lab System
User Guide
4
IF.UG.023 R1
Experimental data.......................................................................................................44
Run Setpoints screen ..................................................................................................46
Overview screen .........................................................................................................49
Instrumentation..........................................................................................................56
Taring ..........................................................................................................................57
Pump Speed ................................................................................................................63
Pressure/Flow .............................................................................................................64
Weights .......................................................................................................................65
Data.............................................................................................................................66
PID...............................................................................................................................67

KrosFlo® TFDF™ Lab System
User Guide
KrosFlo® TFDF™ Lab System
User Guide
5
IF.UG.023 R1
List of tables
Table 1. Explanation of user attention phrases..................................................................................9
Table 2. Explanation of symbols.........................................................................................................9
Table 3. Instrument safety labels .....................................................................................................10
Table 4. ProConnex® TFDF™ Flow Path specifications .....................................................................17
Table 5. Sample and process ............................................................................................................31
Table 6. KrosFlo® TFDF™ Lab System permeate tubing flow rates ..................................................37
Table 7. Parameter data...................................................................................................................62
Table 8. Warning: Product use limitation.........................................................................................69
Table 9. System output.....................................................................................................................70
Table 10.System input.......................................................................................................................70
Table 11. System construction ...........................................................................................................70
Table 12. System environment ..........................................................................................................71
Table 13. System components list......................................................................................................71
List of figures
Figure 1. KrosFlo® TFDF™ Lab System ................................................................................................8
Figure 2. KrosFlo® TFDF™ Lab System Components ........................................................................11
Figure 3. Controller...........................................................................................................................11
Figure 4. Pump Station .....................................................................................................................12
Figure 5. ProConnex® TFDF™ Flow Path...........................................................................................13
Figure 6. Bench space needed..........................................................................................................14
Figure 7. Arranging the system on the bench ..................................................................................14
Figure 8. Assembling the stand ........................................................................................................15
Figure 9. System connections...........................................................................................................16
Figure 10.ProConnex® TFDF™ Flow Path configuration....................................................................17
Figure 11. Connecting pressure sensors ............................................................................................21
Figure 12. Making tubing connections ...............................................................................................22
Figure 13. Touchscreen display ..........................................................................................................23
Figure 14. Main Menu screen ............................................................................................................24
Figure 15. Menu bar...........................................................................................................................24
Figure 16. Screen navigation ..............................................................................................................25
Figure 17. Set-up before priming .......................................................................................................26
Figure 18. Primed system...................................................................................................................28
Figure 19. Experiment workflow ........................................................................................................32
Figure 20. Information screen............................................................................................................33
Figure 21. Main Menu screen ............................................................................................................34
Figure 22. Admin screen.....................................................................................................................35
Figure 23. System Settings screen......................................................................................................36
Figure 24. Reset Tubing Calibration ...................................................................................................37
Figure 25. PID loop settings screen....................................................................................................39
Figure 26. Alarm Set-up screen ..........................................................................................................40
Figure 27. Alarm Reset button ...........................................................................................................41
Figure 28. Alarm History screen.........................................................................................................42
Figure 29. Lock screen message.........................................................................................................42
Figure 30. Lock screen........................................................................................................................43
Figure 31. Data logging screen ...........................................................................................................43
Figure 32. Data Log Files.....................................................................................................................44
Figure 33. Recorded data example ....................................................................................................45
Figure 34. System Mode screen.........................................................................................................45
Figure 35. Concentration Mode Run Setpoints screen ......................................................................46
Figure 36. Concentration/Diafiltration Mode Run Setpoints screen .................................................47

KrosFlo® TFDF™ Lab System
User Guide
KrosFlo® TFDF™ Lab System
User Guide
6
IF.UG.023 R1
Figure 37. Concentration/Diafiltration/Concentration Mode Run Setpoints screen.........................48
Figure 38. Concentration Mode Overview screen .............................................................................50
Figure 39. Concentration/Diafiltration Mode Overview screen ........................................................51
Figure 40. Concentration/Diafiltration/Concentration Mode Overview screen................................52
Figure 41. Starting the Wizard feature...............................................................................................53
Figure 42. Initial screen ......................................................................................................................53
Figure 43. Initial Wizard feature screen .............................................................................................54
Figure 44. Calculation in progress ......................................................................................................54
Figure 45. Manual Mode Overview screen ........................................................................................55
Figure 46. Reset prompt.....................................................................................................................56
Figure 47. Manual Mode Overview screen ........................................................................................57
Figure 48. Manual Mode Run Setpoints screen .................................................................................58
Figure 49. Filter settings.....................................................................................................................58
Figure 50. Filter List screen (left side) ................................................................................................59
Figure 51. Navigation menu ...............................................................................................................59
Figure 52. Filter List screen (right side) ..............................................................................................60
Figure 53. Plot screen.........................................................................................................................61
Figure 54. Pump Speed plot ...............................................................................................................63
Figure 55. Pressure/Flow plot ............................................................................................................64
Figure 56. Weights plot ......................................................................................................................65
Figure 57. Data plot............................................................................................................................66
Figure 58. PID plot..............................................................................................................................67
Figure 59. Magnetic forces caution and warning...............................................................................69

KrosFlo® TFDF™ Lab System
User Guide
KrosFlo® TFDF™ Lab System
User Guide
7
IF.UG.023 R1
Abbreviations
AC Alternating current
CE Conformitée Européenne
C Concentration
D Diafiltration
DV Diafiltration volume
CF Concentration factor
cm Centimeter
FAS Field Application Scientist
Hz Hertz
in Inches
kg Kilogram
lbs Pounds
LMH Liters/Meters2/Hour
LPM Liters per minute
PCV Percent Cell Volume
PE Pressure Sensor
PID Proportional, Integral and Derivative
PSI Pounds per square inch
PV Process variable
rpm Reps per minute
SP Desired setpoint
TFDF Tangential Flow Depth Filtration
TMP Transmembrane Pressure
UL Underwriters Laboratories
VT Volumetric throughput

KrosFlo® TFDF™ Lab System
User Guide
8
repligen.com IF.UG.023 R1
Introduction
The KrosFlo® TFDF™ Lab System provides a complete solution for the separation of cells from media
during cell culture processes. The technology combines hardware, software and a single-use filter to
achieve the filtration result. Please direct questions regarding specific applications of the technology
to your regional sales representative or field application scientist.
This User Guide provides a reference document for your KrosFlo® TFDF™ Lab System and is updated
on a regular basis. For the latest version of the document, please visit www.repligen.com/resources.
It is highly recommended that the installation process be executed by a trained Repligen engineer.
For further support with troubleshooting or process optimization, please contact your local Repligen
Field Application Scientist.
Welcome
Thank you for choosing the KrosFlo® TFDF™ Lab System for your laboratory. This innovative KrosFlo®
TFDF™ Lab System provides a complete solution for the separation of cells from media during cell
culture processes with superior flux performance, scalability and ease of use.
Figure 1. KrosFlo® TFDF™ Lab System

KrosFlo® TFDF™ Lab System
User Guide
9
repligen.com IF.UG.023 R1
About this document
Several user attention phrases are used throughout this manual. Each phrase should draw the
following level of attention:
Table 1. Explanation of user attention phrases
Phrase Description
Note:
Points out useful information
IMPORTANT
Indicates information necessary for proper
instrument operation
CAUTION
Cautions users regarding potentially hazardous
situations in regard to user injury or damage to the
instrument if the information is not heeded
WARNING!
Warns users that serious physical injury can result if
warning precautions are not heeded
Safety precautions
Table 2. Explanation of symbols
Symbol Description
Caution
Risk of danger. Consult Operating Instructions for nature of hazard and
corrective actions. Potentially hazardous situation which, if not avoided, may
result in property/equipment damage
Caution
Risk of crushing. Keep fingers away from rotor while pump is in operation. Stop
pump before loading or unloading tubing
Caution
Hot surface. Do not touch
Caution
Risk of electric shock, consult Operating Instructions for nature of hazard and
corrective actions
Safety
Alert
Symbol
Hazard to personnel is present, the SAS is omitted when the hazard is related to
property/equipment damage only
Danger
Imminently hazardous situation which, if not avoided, will result in death or
serious injury
Warning
Pay attention to the magnetic forces when handling the magnetic levitating
centrifugal pump-head. Avoid other magnets or metal parts as contamination
from physical damage or cracks may arise from the magnetic attraction.
Specifically pay attention to the magnetic forces when handling two pump-
heads at the same time

KrosFlo® TFDF™ Lab System
User Guide
10
repligen.com IF.UG.023 R1
Table 3. Instrument safety labels
Symbol Description
Danger
High voltages exist and are accessible. Use extreme caution when servicing
internal components. Remove power from the pump before any cleaning
operation is started
Warning
Remove power from the pump before attempting any maintenance.
Warnings
Tubing breakage may result in fluid being sprayed from pump. Use appropriate
measures to protect operator and equipment
Turn drive off before removing or installing tubes Fingers or loose clothing
could get caught in drive mechanism
Cautions
Power must be turned off before connecting the external remote-control cable
to prevent damage to the drive
Do not contaminate the lubricant in the container, on the shaft or on the seal
with foreign material. Failure to observe this precaution may result in damage
to the seal and premature failure of the seal
No foreign matter should be allowed under the gasket on the back of the front
plate or under the heads of the screws. Failure to observe this precaution may
result in leakage during washdown of the drive
Caution
To avoid electrical shock, the power cord protective grounding conductor must
be connected to the ground. Not for operation in wet locations as defined by
EN61010-1
Warning
High impeller magnetic field strength
The magnetic levitating centrifugal pump head contains a rotor with a rare
earth magnet with high field strength. Pacemakers may be influenced, and
magnetic forces may lead to contusions. Keep distance between pump and
pacemakers, and handle pump heads with care
Caution
Pay attention to the magnetic forces when handling the magnetic levitating
centrifugal pump head. Avoid other magnets or metal parts as contamination
from physical damage or cracks may arise from the magnetic attraction.
Specifically pay attention to the magnetic forces when handling two pump-
heads at the same time
Caution
Keep fingers away from rotor while pump is in operation. Stop pump before
loading or unloading tubing

KrosFlo® TFDF™ Lab System
User Guide
11
repligen.com IF.UG.023 R1
Figure 2. KrosFlo® TFDF™ Lab System Components
Controller
Users change parameters and monitor processes through the interface of the Controller. A 12-inch
touch screen enables user input, such as a change in pump speed and displays a diagram of the
system for visual analysis. The Wizard feature within the KrosFlo® TFDF™ Software, assists running
an optimum process. Based on just a few user specifications, the KrosFlo® Wizard feature generates
run parameters for simple and accurate operation.
Figure 3. Controller
1. 12” Touchscreen interface
2. Control panel
Pump Station
The Pump Station transfers all liquids during the TFDF™ process. A low shear and digitally controlled
magnetic levitating centrifugal style pump delivers feed stock from the bioreactor vertically through
the lumen of the TFDF™ filter and then back into the bioreactor. The Permeate (top) peristaltic
pump transfers permeate from the filter enclosure to the permeate reservoir. The Diafiltration/Aux
peristaltic pump (bottom) transfers buffer from a diafiltration reservoir to the bioreactor during the
diafiltration stage. The overall process is monitored using multiple sensors:
•Three in-line single-use pressure sensors for feed, permeate and retentate
•A non-invasive, clamp-on ultrasonic retentate flow meter
4
1
2
3
5
1
2
1. Controller
2. Pump Station
3. Scale (for bioreactor)
4. Scale (for permeate)
5. ProConnex® TFDF™ Flow Path

KrosFlo® TFDF™ Lab System
User Guide
12
repligen.com IF.UG.023 R1
8
9
10
11
Optional sensors (purchased separately from the system) include:
•An in-line single-use pressure sensor for a secondary filter
•An in-line single-use pressure sensor for a guard/sterile filter
•An in-line turbidity monitor
Figure 4. Pump Station
1. Pump Station
2. Permeate peristaltic pump
3. Diafiltration/Aux peristaltic pump
4. Magnetic levitation pump
5. Stand mount
6. Flow meter
7. Control panel
8. Tubing guide rod
9. Extension rod
10. Sleeve
11. Filter clamp
TFDF™ Filter
The TFDF™ Filter is a tubular depth filter available in process development to pilot and production
scales. The filter functions within an enclosure with ports that deliver feed stock and remove
retentate and permeate. All filter sizes larger than the trial size are available as ProConnex® TFDF™
Flow Paths only, which may be configured from a library of components. Each ProConnex® TFDF™
Flow Path arrives as a fully closed, irradiated device that is ready-to-use. No flushing of the filter is
required.
1
2
3
4
5
6
7

KrosFlo® TFDF™ Lab System
User Guide
13
repligen.com IF.UG.023 R1
Figure 5. ProConnex®TFDF™ Flow Path
1. CPC AseptiQuik® genderless aseptic connector
2. Retentate tubing
3. Retentate pressure sensor (PE02)
4. Vent port
5. Pinch clamp
6. TFDF™ filter enclosure
7. Permeate pressure sensor (PE03)
8. Feed pressure sensor (PE01)
9. CPC AseptiQuik® genderless aseptic connector
10. Magnetic pump head
11. Ferromagnetic fixation disc
1
9
2
3
4
5
6
7
8
10
11

KrosFlo® TFDF™ Lab System
User Guide
14
repligen.com IF.UG.023 R1
WARNING! The Controller weight is 36 lbs. Two people are recommended to lift the
Controller out of the box and place it on the bench top.
FRONT VIEW
SIDE VIEW
5 ft (150 cm)
Set-up
Space requirements
Figure 6. Bench space needed
System arrangement
Figure 7. Arranging the system on the bench
1. Controller
2. Pump Station
3. Scale (for bioreactor)
4. Scale (for permeate)
1
2
3
4

KrosFlo® TFDF™ Lab System
User Guide
15
repligen.com IF.UG.023 R1
Stand assembly
Figure 8. Assembling the stand
5. Tubing guide rod
6. Extension rod (required only for
108 cm flow path)
7. Filter clamp
8. Sleeve
20 cm flow path
configuration
108 cm flow path
configuration
5
6
8
7
5
6
8
7

KrosFlo® TFDF™ Lab System
User Guide
16
repligen.com IF.UG.023 R1
System cable connections
Figure 9. System connections
Optional connections
1. Connect the 718 Minifast Turbidity sensor cable from the Pump Station to your turbidity
sensor.
2. Connect the longer M12 Eurofast depth station flow meter cable from the Pump Station to
your depth station.
KEY
Required
Optional
Non-cable
Note: Use tear drop to align pins with socket.
Communication cable pins are delicate.

KrosFlo® TFDF™ Lab System
User Guide
17
repligen.com IF.UG.023 R1
ProConnex® TFDF™ Flow Path
Table 4. ProConnex® TFDF™ Flow Path specifications
Type Filter Flow path Flow path Flow path Flow path
Surface Area (cm2)
3
3
150
1500
6000
Recommended
process volume
< 1 L
< 1 L
< 50 L
< 500 L
< 2000 L
Filter length (cm)
20
20
108
108
108
Effective length (cm)
2.2
2.2
108
108
108
Number of tubes
1
1
1
10
40
Figure 10. ProConnex® TFDF™ Flow Path configuration
1. CPC AseptiQuik® genderless aseptic connector
2. Retentate tubing
3. Retentate pressure sensor (PE02)
4. Vent port
5. Pinch clamp
6. TFDF™ Filter enclosure
7. Permeate pressure sensor (PE03)
8. Feed pressure sensor (PE01)
9. CPC AseptiQuik® genderless aseptic connector
10. Magnetic pump head
11. Ferromagnetic fixation disc
1
9
2
3
4
5
6
7
8
10
11
Example ProConnex® TFDF™ flow path with
filter, filter enclosure, tubing, clamps,
pressure sensors, magnetic pump head,
vent filter and genderless AseptiQuik®
connectors. Configurations may vary with
your requirements.

KrosFlo® TFDF™ Lab System
User Guide
18
repligen.com IF.UG.023 R1
Install flow path
Unbox the flow path and
remove the ferromagnetic disc.
20 cm flow path installation
1.
2.
3.
Align pump head.
Pull locking pin and
insert pump head.
Rotate filter up
(locking pin will click).
Note: Remove the ferromagnetic disc
before proceeding to the next step.

KrosFlo® TFDF™ Lab System
User Guide
19
repligen.com IF.UG.023 R1
108 cm flow path installation
Transfer tubing guide rod
to extension rod and raise
sleeve to maximum height.
Transfer extension rod
(with tubing guide rod) to
sleeve and rotate clamp so
that it is out of the way
during installation.
Connect retentate
pressure sensor with
RJ12 extender to PE02
This is much easier to
connect before the
flow path is installed
due to its height.
1.
2.
3.
Note: RJ12 cables are not supplied with the ProConnex®
assemblies and must be purchased separately.
Note: Extender available from Repligen support.

KrosFlo® TFDF™ Lab System
User Guide
20
repligen.com IF.UG.023 R1
Align pump head
Pull locking pin and
insert pump head
Rotate filter up
(locking pin will click)
7. Secure flow path with clamp and raise extension
rod so that guide rod is at retentate tubing height.
Make final adjustments as necessary.
4.
5.
6.