REPLIGEN XCell ATF 2 User manual

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For Use with:
•XCell® ATF 2 Stainless Steel Housing
•XCell® ATF 4 Stainless Steel Housing
XCell® ATF 2 and 4
Stainless Steel Housings
User Guide

KrosFlo® TFDF™ Lab System
User Guide
XCell® ATF 2 and 4 Stainless Steel Housings
User Guide
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The information contained in this document is subject to change without notice.
With respect to documentation accompanying product, Repligen makes no warranty, express or
implied. Any and all warranties related to the documentation accompanying product are expressly
disclaimed. Customer shall refer to the terms and conditions of sale governing the transaction for
any and all warranties for the product.
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.
Products are not intended for diagnostic or therapeutic use or for use in vivo with humans or
animals.
For further information, please contact Repligen Corporation at www.repligen.com.
2023 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
[e-mail]
[phone] (Option 1)
Repligen Corporation
111 Locke Drive
Marlborough, Massachusetts 01752
www.repligen.com

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User Guide
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User Guide
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Contents
6.1.1 Installing the diaphragm .............................................................................................10
6.2.1 XCell® ATF 2 Stainless Steel Housing hollow fiber filter installation...........................11
6.3.1 Filter wetting...............................................................................................................12
6.3.2 Integrity testing pre-autoclaving ................................................................................12
6.3.3 Autoclaving .................................................................................................................12
6.3.4 Integrity testing post-autoclaving...............................................................................12
6.4.1 XCell® ATF 2 Stainless Steel tubing set kits and accessories.......................................13
6.4.2 Tubing set kit for XCell® ATF 2 Stainless Steel Housing connected to glass bioreactor
14
6.5.1 Removing the housing from the bioreactor ...............................................................15
6.5.2 HFF Removal ...............................................................................................................15
6.5.3 Decontaminating the HFF for Disposal .......................................................................15
6.5.4 Pump Air Inlet Filter, O-Rings, Gaskets, and Quick Connects.....................................15
7.1.1 Installing the diaphragm .............................................................................................16
7.1.2 XCell® ATF 4 Stainless Steel Housing hollow fiber filter installation...........................17
7.2.1 Pressure testing ..........................................................................................................18
7.2.2 Filter wetting...............................................................................................................18
7.2.3 Integrity testing pre-autoclaving ................................................................................18
7.2.4 Autoclaving .................................................................................................................18
7.2.5 Integrity testing post-autoclaving...............................................................................18
7.3.1 XCell® ATF 4 Stainless Steel Housing tubing set kits and accessories.........................19
7.3.2 Tubing set kit for XCell® ATF 4 Stainless Steel Housing connected to glass bioreactor
20
7.4.1 Removing the housing from the bioreactor ...............................................................21
7.4.2 HFF Removal ...............................................................................................................21
8.3.1 EU Declaration of Conformity.....................................................................................22
8.3.2 Tube set kits................................................................................................................23

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List of tables
Table 1. Explanation of user attention phrases ......................................................................................8
Table 2. Explanation of symbols .............................................................................................................8
Table 3. Dimensions................................................................................................................................9
Table 4. Suggested scale of operation ....................................................................................................9
Table 5. Headplate fitting part numbers...............................................................................................13
Table 6. XCell® ATF 2 tubing set kit options..........................................................................................13
Table 7. XCell® ATF 2 required accessories...........................................................................................13
Table 8. Headplate fitting part numbers...............................................................................................19
Table 9. XCell® ATF 4 tubing set kit options..........................................................................................19
Table 10. XCell® ATF 4 required accessories.........................................................................................19
Table 11. XCell® ATF 2 spare parts........................................................................................................23
Table 12. XCell® ATF 4 spare parts........................................................................................................24
List of figures
Figure 1. Pressure and exhaust strokes ..................................................................................................8
Figure 2. XCell® ATF 2 Stainless Steel Housing......................................................................................10
Figure 3. Orientation of silicone diaphragm in XCell® ATF 2 Stainless Steel Housing ..........................11
Figure 4. XCell® ATF 2 Stainless Steel Housing Installation of HFM......................................................11
Figure 5. XCell® ATF 2 Stainless Steel Housing connection to glass bioreactor (tri-clamp)..................14
Figure 6. XCell® ATF 2 Stainless Steel Housing connection to glass bioreactor
(AseptiQuik®/ReadyMate™).........................................................................................................14
Figure 7. XCell® ATF 4 Stainless Steel Housing......................................................................................16
Figure 8. Orientation of silicone diaphragm in XCell® ATF 4 Stainless Steel Housing ..........................17
Figure 9. XCell® ATF 4 Stainless Steel Housing connection to glass bioreactor (tri-clamp)..................20
Figure 10. XCell® ATF 4 Stainless Steel Housing connection to glass bioreactor (AseptiQuik®)...........20

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Abbreviations
A2B ATF device to bioreactor
A2C ATF device to controller
ATF Alternating tangential flow
DAC Disposable aseptic connectors
DF Driving force
DP Driving pressure
E Exhaust
FAS Field Applications Specialist
HFF Hollow fiber filter
HFM Hollow fiber module
hr Hour
kD kilodalton
L Liter
Lbs Pounds
m Meter
M Molar
min Minute
mm millimeter
NaOH Sodium hydroxide
P Pressure
PA Pump air hemisphere
PES Polyethersulfone
PL Pump liquid hemisphere
ppm Parts per million
PRV Pressure regulator valve
PS Polysulfone
psi Pounds per square inch
psig Pounds per square inch gauge
SIP Steam in Place
SUB Single-use bioreactor
µm micron
California Proposition 65 Warning
Warning
This product can expose you to chemicals including lead, which is
known to the State of California to cause cancer and birth defects or
other reproductive harm. For more information, go to
www.P65Warnings.ca.gov/product.

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1. Introduction
XCell® ATF Technology uses alternating tangential flow (ATF) to intensify upstream processes by
retaining cells in suspension cultures, such as mammalian cell culture and viral vectors. An
innovative diaphragm pump creates alternating tangential flow, leading to high viable cell densities
and increased throughput with lower cost of goods. Typical applications include intensification of
the following processes:
•N-1 fed-batch
•Long-term continuous processing
•Vaccine and virus production
•Gene therapy and media exchange
XCell® ATF 2 and XCell® ATF 4 Stainless steel Housings enable production of high density, high
viability cell culture applications with linear flux and low shear for scale-up to 50 L. The stainless
steel product line includes the housings, XCell® Lab Controllers, software, tubing set kits, pressure
sensors, and accessories.
Hollow fiber filters (HFF) used in XCell® ATF Technology are made of Polyethersulfone (PES) or
polysulfone (PS) and are available in 0.2 and 0.5 µm and 50 kD pore size and 1 mm inner diameter
and are scalable across device sizes.
Single-use tubing set kits and accessories are available for the connection of XCell® ATF 2 and 4
Stainless Steel Housings to both glass and single-use bioreactors, simplifying preparation and
connectivity and allowing for the proper exchange of cell culture material. XCell® Lab Controllers
operate the diaphragm pumps. The housings and tubing set kits are also compatible with legacy C24
Controllers. Additional information on the use of the controllers and XCell® ATF Technology can be
found in XCell® Lab Controller User Guide.
This document describes the major components, set-up, connectivity, and preparation of the glass
XCell® ATF 2 and 4 Housings and hollow fiber filters. Users should possess a basic level of skill in the
areas of aseptic technique and fluid handling.
For further support in optimizing or troubleshooting, please contact your local Repligen Field
Applications Specialist (FAS).

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2. Description of XCell® ATF Stainless Steel Housings
XCell® ATF Technology includes two sizes of housings to support a fully scalable process. An
integrated diaphragm pump provides alternating tangential flow through a hollow fiber filter which
retains large components (i.e., cells) and removes waste or product components (permeate/filtrate).
An additional pump continuously removes cell-free permeate from the system.
2.1 Components
•Filter housing: XCell® ATF 2 utilizes a fully encapsulated hollow fiber membrane that is
attached above the stainless steel diaphragm pump. XCell® ATF 4 includes a stainless steel
housing that holds the hollow fiber filter.
•Diaphragm pump: spherical housing in which a silicone diaphragm membrane is inflated and
deflated by pressurized air or vacuum, creating alternating flow.
•XCell® ATF to Bioreactor (A2B) connection assembly: tubing assembly connecting the
filtration assembly to a bioreactor or process vessel.
•Bioreactor adapter: adapter between the connection assembly and bioreactor port. Typical
fittings for glass bioreactors include Ingold or tri-clamp. Single-use bioreactors (SUB) use
disposable, aseptic connectors.
Note: Bioreactor adapter is not provided.
•Hollow fiber membrane (HFM) or hollow fiber filter (HFF): 0.2 or 0.5 µm or 50 kD pore size
filter, positioned between the bioreactor and the diaphragm pump.
•Filtrate/harvest tubing: sterile tubing and fittings connecting the housing to the collection
vessel. Compatible with a filtrate/harvest pump.
Note: Filtrate/harvest tubing and pump are not provided.
2.2 XCell® ATF Housing pump cycle
The diaphragm pump generates alternating tangential flow (ATF) through the hollow fiber filter. ATF
is a pulsating, reversible flow of liquid between a process vessel and diaphragm pump.
The process consists of two cycles (Figure 1), the pressure cycle (P-cycle) and the exhaust cycle (E-
cycle). The P cycle occurs when air is fed into the ATF pump, moving the diaphragm up towards the
pump liquid-side (PL) hemisphere and driving the liquid from the diaphragm pump through the HFF
and into the process vessel.
The E-cycle occurs when vacuum is introduced, thus pulling the diaphragm down towards the pump
air-side (PA) hemisphere, and liquid is pulled from the process vessel through the HFF, and back into
the diaphragm pump. The diaphragm must travel between the two extremes to complete one cycle.
The flow through the hollow fiber filter generates tangential flow in each direction in an alternating
fashion.

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Figure 1. Pressure and exhaust strokes
Note: XCell® ATF Stainless Steel Housings require a vacuum (negative pressure) to move the
diaphragm to its lowest position. Positive pressure from the bioreactor is insufficient to
completely deflate the diaphragm, necessitating the use of vacuum to ensure proper
XCell® ATF Stainless Steel Housing operation.
3. 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 regarding 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
4. 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
Safety
Alert
Symbol
Hazard to personnel is present, the safety alert symbol is omitted when the
hazard is related to property/equipment damage only
From bioreactor
To bioreactor
Filtered harvest
HFF
HFF
Diaphragm
Diaphragm
Positive air pressure
Negative air pressure
Filtered harvest

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5. Specifications
5.1 Utility requirements
Utility requirements for the XCell® ATF 2 and 4 Systems can be found in the XCell® Lab Controller
User Guide.
Table 3. Dimensions
Filtration Assembly Dimensions1H, W Nominal footprint (m2)
XCell® ATF 2 SS Pump Housing
30", 5"
0.04
XCell® ATF 4 SS Pump Housing
21", 8"
0.13
1Indicated dimensions are estimates for the filtration assembly, as the choice of cartridge and the
connection to the bioreactor and accessories can affect height and effective area.
Housing selection depends on the perfusion rate or total volume throughput in addition to other
process conditions (Table 4).
Table 4. Suggested scale of operation
Housing Bioreactor size
XCell® ATF 2 2 - 10 L
XCell® ATF 4 10 - 50 L

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6. XCell® ATF 2 Stainless Steel Housing
XCell® ATF 2 Stainless Steel Housing (Figure 2) is recommended for use with 2 - 10 L bioreactors. The
assembly of the housing and accessories includes installing the diaphragm into the housing,
connecting the hollow fiber filter to the housing, and connecting the filter to the bioreactor and the
harvest vessel. The system is prepared for use by filter wetting, integrity testing, and autoclaving.
Figure 2. XCell® ATF 2 Stainless Steel Housing
6.1 Stainless steel XCell® ATF 2 Housing assembly
XCell® ATF 2 Stainless Steel Housing assembly includes installation of the diaphragm and hollow fiber
filter and connection of the tubing from the housing to the bioreactor and the housing to the harvest
vessel.
6.1.1 Installing the diaphragm
Important: The silicone diaphragm must be replaced after each run and prior to sterilization. To
avoid premature failure, it is important to ensure that the diaphragm is seated in the correct
orientation. Use with aqueous liquids only. For use with other fluids contact Repligen for a technical
discussion. When used as instructed, the diaphragm is designed to last over one million
pressure/exhaust cycles.
1. Hollow fiber membrane
2. Permeate/harvest
3. Retentate
4. Diaphragm pump
5. Air inlet
6. Drain port

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Figure 3. Orientation of silicone diaphragm in XCell® ATF 2 Stainless Steel Housing
6.2 XCell® ATF 2 Device Diaphragm Placement
Important: The silicone diaphragm must be replaced after each run, prior to sterilization.
1. Set the pump air-side (PA) hemisphere (Figure 3, 2) on a flat surface with the open (concave)
side of the hemisphere facing up.
2. Place diaphragm (3) with the convex side oriented up onto the PA hemisphere, and place
the PL hemisphere, wide opening down, on top of the diaphragm.
3. Assure the diaphragm gasket O-ring is positioned uniformly around the periphery of the
pump O-ring groove. Place the pump liquid-side (PL) hemisphere (1) on top. The two
hemispheres, with diaphragm in between, should be equally spaced.
4. Clamp (4) the two hemispheres together.
5. Secure the diaphragm pump to the stand.
6.2.1 XCell® ATF 2 Stainless Steel Housing hollow fiber filter installation
XCell® ATF 2 Stainless Steel Housing accommodates a variety of hollow fiber modules. For selection
support, please contact your local Repligen Field Applications Specialist (FAS).
Figure 4. XCell® ATF 2 Stainless Steel Housing Installation of HFM
1. Pump liquid-side hemisphere
2. Pump air-side hemisphere
3. Diaphragm
4. Clamp
1. Hollow fiber module (not included)
2. Clamp
3. Gasket
4. PL hemisphere
5. Drain port

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1. Place a sanitary gasket (Figure 4, 3) into the sanitary groove on top of the Pump Liquid-side
(PL) hemisphere (4).
2. Connect the HFM (1) to the pump housing by aligning it with the sanitary gasket on the top
of the PL hemisphere (4).
3. Secure the connection tightly with a ¾" tri-clamp (2).
6.3 Filter preparation
To prepare the filter for use, the assembled housing should be wetted, integrity tested and
autoclaved.
6.3.1 Filter wetting
Hollow fiber filters require wetting after installation and pressure testing. Slight variations in
assembly are possible depending on the configuration of equipment purchased and its intended use.
Refer to the XCell® ATF Technology Filter Preparation and Autoclaving User Guide for instructions.
6.3.2 Integrity testing pre-autoclaving
Note: Repligen strongly recommends the practice of pre-autoclaving integrity testing to ensure a
proper assembly prior to autoclaving.
Refer to the XCell® ATF Technology Filter Preparation and Autoclaving User Guide for configurations
for both glass and single-use bioreactor integrity tests and testing instructions and criteria.
6.3.3 Autoclaving
The XCell® ATF 2 Stainless Steel Housing assembly is autoclaved either before or after integrity
testing. XCell® ATF 2 comprises a small-diameter, self-contained hollow fiber filter connected to a
diaphragm pump, and can be autoclaved using a pre-programmed liquid cycle at 121°C for 30
minutes. Refer to the XCell® ATF Technology Filter Preparation and Autoclaving User Guide for
instructions.
6.3.4 Integrity testing post-autoclaving
Post-autoclaving integrity testing is required only if testing was not performed pre-autoclaving. Refer
to the XCell® ATF Technology Filter Preparation and Autoclave User Guide for instructions.
6.4 Connecting XCell® ATF 2 Stainless Steel Housing to a bioreactor
XCell® ATF 2 Stainless Steel Housing is generally connected to the bioreactor prior to autoclaving.
Fluid management for XCell® ATF 2 Stainless Steel Housings include retentate (A2B) tubing set kits
and accessories that connect the housing with the bioreactor, ensuring proper exchange of cell
culture material. The A2B tubing connects to XCell® ATF 2 with a tri-clamp. The bioreactor end of the
tubing is available with either a tri-clamp or an AseptiQuik® or Readymate™ disposable aseptic
connector. The tubing is connected to the headplate (Table 5) and feeds into a dip tube (P/N TC-DIP-
LAB-.25/.5-250 or TC-DIP-.25/.5-250. Permeate tubing connects to the housing with a hose barb and
should be sterilized by autoclave or attached to the harvest vessel with a tubing welder or
disposable sterile coupling. The permeate tubing should be compatible with the permeate/harvest
pump. Tubing is also provided for the bottom permeate and drain ports, both of which connect with
a hose barb.

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Table 5. Headplate fitting part numbers
Head plate fitting Bioreactor
HP:MPP-19 3 L Applikon
HP:MPP-M26 5 - 10 L Applikon
HP:BBI-M26 Sartorius/BBI
HP-PG13.5L Millipore Mobius® CellReady
PG13.5 Generic
6.4.1 XCell® ATF 2 Stainless Steel tubing set kits and accessories
Several components are required to connect the XCell® ATF 2 Stainless Steel Housing to a bioreactor.
XCell® ATF 2 autoclavable tubing set kits (Table 5) are available in three configurations depending on
the type of bioreactor in use and the preferred connectors. All tubing set kits have a pressure rating
of 25 psi. Additional accessories (Table 6) are required, regardless of the chosen tubing set.
Table 6. XCell® ATF 2 tubing set kit options
Tube set part number Tubing connection Use
ATF2-A2B-TC Tri-clamp/tri-clamp ATF to bioreactor
ATF2-A2B-RMTC GE ReadyMate™/tri-clamp ATF to bioreactor
ATF2-A2B-TCAQ AseptiQuik®/tri-clamp ATF to bioreactor
ATF2-A2B-AQ AseptiQuik®/ AseptiQuik® ATF to bioreactor
Table 7. XCell® ATF 2 required accessories
Description Part number Single/Multi-use
Pressure
rating
ID
Material of
construction
Top and bottom
permeate and drain
SSATF2-TSK SU 25 psi 0.25”
Platinum-
cured silicone

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6.4.2 Tubing set kit for XCell® ATF 2 Stainless Steel Housing connected to glass bioreactor
Assembly of the housing, tubing set, and accessories is completed prior to autoclaving.
Figure 5. XCell® ATF 2 Stainless Steel Housing connection to glass bioreactor (tri-clamp)
A. TC-DIP-LAB-.25/.5-250 or
TC-DIP-.25/.5-250 (Dip tube)
B. ATF2-A2B-TC (A2B)
C. SSATF2-TSK
C1. Top permeate
C2. Bottom permeate
C3. Drain
Connection instructions (Figure 5):
1. Ensure dip tube (A) is attached to head plate of bioreactor.
2. Attach A2B tubing (B) to XCell® ATF 2 and bioreactor head plate.
3. Attach top permeate (C1) to hose barb.
4. Attach bottom permeate (C2) to hose barb.
5. Attach drain (C3) to hose barb. Autoclave.
Figure 6. XCell® ATF 2 Stainless Steel Housing connection to glass bioreactor
(AseptiQuik®/ReadyMate™)
A. TC-DIP-LAB-.25/.5-250 or
TC-DIP-.25/.5-250 (Dip tube)
B. ATF2-A2B-RMTC or
ATF2-A2B-TCAQ or
ATF2-A2B-AQ (A2B)
C. SSATF2-TSK
C1. Top permeate
C2. Bottom permeate
C3. Drain
Connection instructions (Figure 6):
1. Ensure dip tube (A) is attached to head plate of bioreactor.
2. Attach A2B tubing (B) to XCell® ATF 2 and bioreactor head plate.
Note: Bioreactor adapter tubing not provided.
3. Attach top permeate (C1) to hose barb.

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4. Attach bottom permeate (C2) to hose barb.
5. Attach drain (C3) to hose barb. Autoclave.
6.5 Disassembly and maintenance
6.5.1 Removing the housing from the bioreactor
1. Stop the permeate/harvest pump and disconnect the harvest bag from the housing.
2. Stop and disconnect the A2C line from the diaphragm pump.
3. Close or disconnect compressed air and vacuum services from the pneumatic enclosure.
4. Disconnect sensors from the filtration assembly to pneumatic enclosure.
5. If necessary, remove the pneumatic enclosure from the filtration assembly.
6. Close valves connecting the filtration assembly to the vessel.
7. Drain liquid from the connection.
8. Optionally, purge the connection with water, steam, or some other medium.
9. Disconnect the filtration assembly from vessel.
10. Remove the filtration assembly to an appropriate work area.
6.5.2 HFF Removal
1. Drain the system into an appropriate waste vessel or drain.
2. Remove all sensors from the housing.
3. Remove air inlet filter connected to the diaphragm pump and any other connections to the
XCell® ATF Housing.
4. Open the clamp connecting the filter to the diaphragm pump and separate.
5. Open the clamp connecting the filter to the A2B tubing and separate.
6. Remove the O-ring at the exposed end of the HFF.
7. Press the HFF in the opposite direction to remove.
8. Prepare filter housing for cleaning and reuse.
6.5.3 Decontaminating the HFF for Disposal
1. Rinse the HFF two times with water to remove residual cells and debris.
2. Decontaminate with either 0.5M NaOH, potassium hydroxide, or bleach for 2 – 24 hours.
Note: 5.25% chlorine is equivalent to 52,500 parts per million (ppm). If household chlorine bleach is
used, two working solutions are generally recommended: 5000 - 6000 ppm for initial
decontamination of organic spill material, and 500 - 600ppm for disinfection of cleaned
surfaces.
6.5.4 Pump Air Inlet Filter, O-Rings, Gaskets, and Quick Connects
All consumable items are single use.

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7. XCell® ATF 4 Stainless Steel Housing
XCell® ATF 4 Stainless Steel Housing (Figure 7) is recommended for use with 10 – 50 L bioreactors.
The assembly of the housing and accessories includes installing the diaphragm and hollow fiber filter
into the housing, connecting the tubing set kit to the bioreactor, and connecting the harvest line to
the housing. The system is prepared for use by pressure testing, filter wetting, integrity testing, and
autoclaving.
Figure 7. XCell® ATF 4 Stainless Steel Housing
7.1 XCell® ATF 4 Stainless steel Housing assembly
XCell® ATF 4 Stainless Steel Housing assembly includes installation of the diaphragm and hollow fiber
filter and assembly of the tubing set kit that will connect the housing and the bioreactor and the
tubing that will connect the housing and the harvest vessel.
7.1.1 Installing the diaphragm
Important: The silicone diaphragm must be replaced after each run and prior to sterilization. To
avoid premature failure, it is important to ensure that the diaphragm is seated in the correct
orientation. Use with aqueous liquids only. For use with other fluids contact Repligen for a technical
discussion. When used as instructed, the diaphragm is designed to last over one million
pressure/exhaust cycles.
1. Housing
2. Permeate/Harvest
3. Retentate
4. Diaphragm pump
5. Air inlet
6. Drain Port

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Figure 8. Orientation of silicone diaphragm in XCell® ATF 4 Stainless Steel Housing
7.1.1.1 XCell® ATF 4 Stainless Steel Housing diaphragm placement
1. Set the pump air-side (PA) hemisphere (Figure 8, 2) on a flat surface with the open (concave)
side of the hemisphere facing up.
2. Place diaphragm (3) with the convex side oriented up onto the PA hemisphere, and place
the PL hemisphere, wide opening down, on top of the diaphragm.
3. Assure the diaphragm gasket O-ring is positioned uniformly around the periphery of the
pump O-ring groove. The two hemispheres, with diaphragm in between, should be equally
spaced.
4. Clamp (4) the two hemispheres together.
5. Secure the diaphragm pump to the stand.
7.1.2 XCell® ATF 4 Stainless Steel Housing hollow fiber filter installation
XCell® ATF 4 Stainless Steel Housing accommodates a variety of hollow fiber filters. For support in
selecting hollow fiber filters, please contact your local Repligen Field Applications Specialist (FAS).
1. Stand the filter housing vertically, with harvest port up.
2. Insert the HFF into the housing.
3. Insert one O-ring into the groove between HFF and housing end-ferrule wall.
4. Place the reducer onto the end, forcing the O-ring into the groove.
5. Clamp the reducer to the housing, compressing the O-ring against the HFF. A seal is formed
between the housing, reducer and HFF.
6. On the other end of the filter housing, place the second O-ring into the groove between HFF
and housing end-ferrule.
7. Carefully place the filter housing onto the diaphragm pump, forcing the O-ring deeper into
the groove.
8. Clamp the filter housing to the diaphragm pump.
9. Assemble the rest of the XCell® ATF Housing, including tubing set kits.
7.2 Filter preparation
To prepare the filter for use, the assembled housing should be pressure tested, wetted, integrity
tested and autoclaved.
1. Housing assembly, pump liquid-side
(PL) hemisphere
2. Housing assembly, pump air- side
(PA) hemisphere
3. Diaphragm
4. Clamp

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7.2.1 Pressure testing
Pressure testing ensures that the XCell® ATF 4 Stainless Steel Housing assembly is free of leaks and
can withstand the required operating pressures. The assembly can be pressure tested with or
without a filter module, before or after sterilization. Ideally, the test is performed with a filter prior
to autoclaving. Slight variations in the configuration of the external connections are possible
depending on the configuration of the equipment purchased and the connections being made to the
bioreactor. Refer to the XCell® ATF Technology Filter Preparation and Autoclaving User Guide for
instructions.
7.2.2 Filter wetting
Hollow fiber filters require wetting after installation and pressure testing. Slight variations in
assembly are possible depending on the configuration of equipment purchased and its intended use.
Refer to the XCell® ATF Technology Filter Preparation and Autoclaving User Guide for instructions.
7.2.3 Integrity testing pre-autoclaving
Note: Repligen strongly recommends the practice of pre-autoclaving integrity testing to ensure a
proper assembly prior to autoclaving.
Refer to the XCell® ATF Technology Filter Preparation and Autoclave User Guide for configurations
for both glass and single-use bioreactor integrity tests and testing instructions and criteria.
7.2.4 Autoclaving
The XCell® ATF 4 Stainless Steel Housing assembly is autoclaved either before or after integrity
testing. The cycle sequence presented is strongly recommended to ensure sterility and to retain
filter integrity and performance. The procedure includes three pre-vacuum pulses, which provide
proper chamber evacuation for a faster and more consistent ramp to the required temperature.
Refer to the XCell® ATF Technology Filter Preparation and Autoclaving User Guide for instructions.
7.2.5 Integrity testing post-autoclaving
Post-autoclaving integrity testing is required only if testing was not performed pre-autoclaving. Refer
to the XCell® ATF Technology Filter Preparation and Autoclave User Guide for instructions.
7.3 Connecting XCell® ATF 4 Stainless Steel Housing to a bioreactor
Fluid management for XCell® ATF 4 Stainless Steel Housings include retentate (A2B) tubing set kits
and accessories that connect the housing with the bioreactor, ensuring proper exchange of cell
culture material. The A2B tubing connects to XCell® ATF 4 with a tri-clamp. The bioreactor end of the
tubing is available with either a tri-clamp or an AseptiQuik® disposable aseptic connector. The tubing
is connected to the headplate (Table 8) and feeds into a dip tube (P/N TC:DIP-10/250 or TC:DIP-
10/450). A pressure sensor kit connects to the housing with a tri-clamp and the permeate tubing
connects to the pressure sensor. The permeate tubing should be sterilized by autoclave or attached
to the harvest vessel with a tubing welder or disposable sterile coupling. The permeate tubing
should be compatible with the permeate/harvest pump. Tubing is also provided for the drain port
and connects with a hose barb.

XCell® ATF 2 and 4 Stainless Steel Housings
User Guide
19
SG-SSATF24 rev 1
Table 8. Headplate fitting part numbers
Head plate fitting Bioreactor
HP:MPP-19
3 L Applikon
HP:MPP-M26
5 -10 L Applikon
HP:BBI-M26
Sartorius/BBI
HP-PG13.5L
Millipore Mobius® CellReady
PG13.5
Generic
7.3.1 XCell® ATF 4 Stainless Steel Housing tubing set kits and accessories
Several components are required to connect the XCell® ATF 4 Stainless Steel Housing to a bioreactor.
XCell® ATF 4 autoclavable tubing set kits (Table 9) are available in three configurations depending on
the type of bioreactor in use and the preferred connectors. All tubing set kits have a pressure rating
of 25 psi. Additional accessories (Table 10) are required, regardless of the chosen tubing set.
Table 9. XCell® ATF 4 tubing set kit options
Tube set part number Tubing connection
ATF4-A2B-TC Tri-clamp
ATF4-A2B-TCAQ AseptiQuik®
Table 10. XCell® ATF 4 required accessories
Description Part number
Single- or
multi-use
Pressure
rating
ID
Material of
construction
Top permeate and drain SSATF4-TSK SU 25 psi 1/4”
Platinum-cured
silicone
Dip tube
TC:DIP-10/250 or
TC:DIP-10/450
MU N/A 3/8” 316 L SS
Pressure sensor kit SSATF4-PSK-V2 SU 25 psi 3/8” Polycarbonate

XCell® ATF 2 and 4 Stainless Steel Housings
User Guide
20
SG-SSATF24 rev 1
7.3.2 Tubing set kit for XCell® ATF 4 Stainless Steel Housing connected to glass bioreactor
Assembly of the housing, tubing set, and accessories is completed prior to autoclaving.
Figure 9. XCell® ATF 4 Stainless Steel Housing connection to glass bioreactor (tri-clamp)
A. TC:DIP-10/250 or TC:DIP-10/450 (dip tube)
B. ATF4-A2B-TC (A2B)
C. SSATF-PSK-V2 (pressure sensor)
D. SSATF4-TSK
D1. Top permeate
D2. Drain
Connection instructions (Figure 9):
1. Ensure dip tube (A) is attached to head plate of bioreactor.
2. Attach A2B tubing (B) to XCell® ATF 4 and bioreactor head plate.
3. Connect pressure sensor (C) to XCell® ATF 4.
4. Attach top permeate (D1) to pressure sensor (C).
5. Attach drain (D2) to hose barb. Autoclave.
Figure 10. XCell® ATF 4 Stainless Steel Housing connection to glass bioreactor (AseptiQuik®)
A. TC:DIP-10/250 or TC:DIP-10/450 (dip tube)
B. ATF4-A2B-TCAQ (A2B)
C. SSATF-PSK-V2 (pressure sensor)
D. SSATF4-TSK
D1. Top permeate
D2. Drain
Connection instructions (Figure 10):
1. Ensure dip tube (A) is attached to head plate of bioreactor.
2. Attach A2B tubing (B) to XCell® ATF 4 and bioreactor head plate.
Note: Bioreactor adapter tubing not provided.
3. Connect pressure sensor (C) to XCell® ATF 4.
4. Attach top permeate (D1) to pressure sensor (C).
5. Attach drain (D2) to hose barb. Autoclave.
7.4 Disassembly and maintenance