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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SG-SSATF24 rev 1
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

KrosFlo® TFDF™ Lab System
User Guide
XCell® ATF 2 and 4 Stainless Steel Housings
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

KrosFlo® TFDF™ Lab System
User Guide
XCell® ATF 2 and 4 Stainless Steel Housings
User Guide
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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

KrosFlo® TFDF™ Lab System
User Guide
XCell® ATF 2 and 4 Stainless Steel Housings
User Guide
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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