USD3117c

Allegro™ STR Single-Use Stirred Tank Bioreactors

Redefining single-use cell culture performance

The Allegro STR bioreactor family combines Pall’s uT
 he single bottom-mounted impeller design
bioprocess engineering expertise, cell culture allows the biocontainer to be packed into a
know-how and our drive for quality into a series of compact unit that can be placed directly into the
single-use bioreactors that deliver consistent and hardware with minimal operator manipulation
scalable cell culture performance across the range.
u Installation and inflation of the biocontainer is
The Allegro STR bioreactor family has a direct
achieved in < 30 minutes through a guided
bottom-driven impellor allowing power inputs
sequence via the Human Machine Interface (HMI)
up to 0.25 W/kg, resulting in high oxygen transfer
rates and short mixing times. uT
 he low aspect ratio of the systems minimize the
operations that have to be performed at height
From the outset of the design, our technology
placed strong emphasis toward providing a uA
 utomatic biocontainer deflation speeds up
compact, ergonomic and intuitive turn key bioreactor turnaround time, minimizes operator activity
design concept to maximize usability and process and reduces the overall volume of waste
assurance, while maintaining optimal performance for disposal1
needed in a cell culture environment through several
easy and intuitive operation features:
All product contact surfaces in the Allegro STR bioreactors u Typical mixing data for the Allegro STR bioreactor range:
are single-use components therefore the demands on
maintenance, cleaning and cleaning validation are minimal. 18

Mixing Time (Seconds)

This makes single-use bioreactors the first choice for process 16
development, CMO plants or multiproduct manufacturing 14
12
facilities where maximum flexibility is required.
10
8
Extensive experiments were performed to characterize the 6
bioreactor and gather data on mixing, mass transfer and cell 4
culture performance. Additionally, all components of this 2
10.3 11.4 13.5 16.0
bioreactor were rigorously tested, and several long term 0
50 L 200 L 1000 L 2000 L
experiments were completed in order to ensure the integrity
and compatibility of the materials with regards to cell culture
uM
 ixing and kLa data demonstrate that the systems
demands. During the product development, 45 day life tests
offer a broad design space allowing for good cell
were performed under maximum operating conditions to prove the
culture performance for a wide range of mammalian
design of the single-use system: no leaks were observed during
cell culture processes.
this testing. A smaller number of life tests were also performed
for >100 days continuous operation with no leaks observed. Cell Culture Performance and Scalability:
2.0 x 107 100%

Viable Cells (cells.mL-1)

Integrated testable gamma 1.6 x 107 80%
irradiated Emflon® II vent
filter with heating jacket

Cell Viability
1.2 x 107 60%
Well structured graphical
user interface
8.0 x 106 40%

Siemens PLC based

controller with 6 digital 4.0 x 106 20%
mass flow meters

Large viewing window and 0 0%

transparent film allow visual 0 48 96 144 192
Time (Hours)
checks on cell culture
Viable Cells
Ports for aseptic insertion 10 L 10 L 200 L 1000 L
of conventional probes and Cell Viability
single-use sensors 10 L 10 L 200 L 1000 L

Please refer to Pall document USD3135 ‘Cultivation of Chinese Hamster

Ovary (CHO) Cells in Allegro STR 1000 Single-Use Stirred Tank
Features and Benefits Bioreactor System’ for further details.

Performance uG
 eometric similarity across the product range ensures scalable
Mixing and kLa Performance: performance across the Allegro STR bioreactor family

 igh kLa (up to 40 h-1) and short mixing times (< 30 seconds)
uH uS
 ingle-use pH and DO sensors provide equivalent
achieved by bottom mounted direct drive, pitched blade performance to conventional (control) sensors
impeller.
100 7.30
u Typical kLa data for the STR bioreactor range: 90 7.25
Agitation Speed kLa Range 80 7.20
DO (%)

70 7.15
Allegro STR Scale (L) Range (rpm) (h-1)
pH

60 7.10
50 25 – 200 0.31 – 24.2 50 7.05
200 25 – 150 2 – 33 40 7.00
30 6.95
1000 25 – 105 4 – 41
20 6.90
2000 25 – 105 7.5 – 29.5 0 48 96 144 192
Time (Hours)
DO Control DO Single-use pH Control pH Single-use

2
Usability Intuitive Operation
Simple Installation and Inflation u Totally Integrated Siemensu Automation Portal (TIA Portal)
software platform with standard TFT color touch screen Panel
u Biocontainer easy to unpack and carry up to 2000 L scale
PC (SIMATIC IPC 477D) enables ease of navigation while
uF
 olded biocontainer easily placed directly onto floor of configuring setup or running am operation
bioreactor hardware, no operator intervention to the film
uE
 asy to set up and run local control time based pump
during installation
profiling to support various feeding strategies
uS
 tep-by-step instructions and color-coded numbered labels
uA
 larms and status clearly identified with full audit trail available
guide the operator through the processes
in a batch report
uM
 ulti-point biocontainer directional arrows to ensure correct
uE
 asy to establish different local cascades to maintain
installation, and mitigate operator error during installation
bioreactor control strategies to support culture demands
u Installation and inflation of the biocontainer in less than
uD
 ynamic processing flexibility provided through either weight
30 minutes
or time profile feeding features
u Only one operator for most manipulations
u Clear to user when operator intervention is required
uA
 utomatic inflation sequence is enabled through self guided
lifting arm1 and preconfigured tubing management system
uP
 ackaging unique rip cord design in packaging design to
alleviate need for sharps being used setup and installation

Working at Height Minimized

uE
 ntire portfolio maintains low aspect ratio to reduce overall
system height and increase efficiency on CO2 strip rates
uE
 asy access from floor level to entire bioreactor at all
volume sizes
u All Allegro STR bioreactors, including the 2000 L, height < 3 m
u Reduced risk of operator injury due to shortened height

1 Available only on the Allegro STR 1000 and 2000 L bioreactors.

www.pall.com 3
Process Assurance and Reliability Sensors

Biocontainer Packaging uC
 an be operated with single-use sensors (pH and DO) and /
or up to six conventional probes
u Compact and protective packaging
uA
 dditional analogue connections for additional sensor
u Integral components provide support for biocontainer film, integration (one sensor connection is configured for
tubing and connectors during both installation and use dissolved carbon dioxide probe input)
uP
 rotects the biocontainer and components during u High flexibility in process measurements
transportation and installation
uS
 ensor calibration includes incorporated guided
prompt sequence to ensure calibration is performed
for batch consistency
uA
 llows integration of various process analytical measurements
for process automation
uP
 rocess mitigation steps enabled through sensor redundancy
available for both pH and DO sensor technology

Allegro STR 1000 bioreactor Allegro STR 50 bioreactor

Installation and Inflation

u Procedures predominantly automated
uB
 ioreactor microswitches provide operator feedback
of correct step completion
uM
 inimizes operator intervention and prevents
accidental damage
u Reduced risk for out of box failures
uO
 perator cannot advance to inflation sequence until correct
installation is confirmed by the HMI (controlled via interlocks
Control Software
and switches on the hardware)
uV
 alidated control software, can be connected via OPC
UA server to SCADA (supervisory control and data
acquisition) system
u Developed in accordance with GAMP5
uE
 asy and safe operation in stand-alone mode or integrated
into supervisory system

4
Applications Bioreactor Hardware
Typical applications include: The bioreactor hardware provides the support for the
biocontainer. It has been designed with a viewing window to
uM
 ammalian suspension cell culture, using e.g. CHO or
observe the cell culture at any time. Heating is provided via a
HEK293 cells
water jacket on all Allegro STR bioreactors. On the Allegro STR
u Insect cell culture, using e.g. Sf9 cells 200 bioreactor, an option with a heating mat is also available.
Pall offers a Temperature Control Unit (TCU) to work with the
u Suspension cells in batch, fed-batch and perfusion mode Allegro STR bioreactors and the integration of other common
u Adherent cell growth on microcarriers, ex. VERO TCUs can be discussed. The Allegro STR bioreactor portfolio
also can be equipped to communicate with third party TCUs.
The bioreactor hardware has integrated load cells to provide
Engineering Characterization information on the liquid mass.
Pall has conducted a range of calculations and experiments to
characterize the Allegro STR bioreactors. Mixing characterization
testing methods, such as mixing time, fluid velocity profiles
and Kolmogorov scale were assessed in Computational Fluid
Dynamics studies (CFD). The mixing model was then verified
in a tracer experiment. Other important parameters like kLa,
dissolved CO2 removal and heating/cooling profiles with
thermocouples inserted were also performed. The experimental
setup and results are documented in a separate application note.

Geometry
When filled with the maximum working volume, the Allegro STR
bioreactors have an aspect ratio of approximately (1.4:1).
This significantly reduces the overall height of the bioreactor
compared to cylindrical versions of the same working volume.
It is also much easier and safer for the operators to perform most
installation steps without the use of ladders, hoists, or platforms.
The filled biocontainer in the bioreactor forms in a cubical design.
The cubical design facilitates mixing as natural baffles and
increase the turbulence in the bioreactor. Additional baffles have
been placed at three sides of the container to further increase
that effect, especially at the high end of agitation speed. Pall has
Controller
performed extensive studies using CFD to show that there are The controller uses a Siemens TIA Portal platform with a color
no dead zones and 95% homogenization can be reached within touch screen Panel PC (SIMATIC IPC 477D) HMI. The controller
less than 16 seconds (full impeller speed at up to 2000 L can connect to higher networks via OPC.
working volume).
Operator interaction with the bioreactor is via a large HMI color
screen. Industrial design guidelines were used to provide all the
information in an intuitive and well-arranged format on screen.

www.pall.com 5
Gas Flow Control uS
 pare analog inputs (4-20 mA signal) can be used to connect
external pumps, balances and sensors without modifications
All biocontainers have the option to provide gas through the on the hardware. The ancillary inputs added also are
sparger, into the head space overlay and into a separate open configured on the HMI
pipe sparger for additional CO2 removal. The controller hosts 6
uS
 ingle-use sensors for pH and DO are available
digital thermal mass flow controllers to control the gas flow.
on selected models
Four of them are calibrated to assure the proper mix of air,
oxygen, nitrogen and carbon dioxide through the main sparger.
Another is calibrated for air on the CO2 removal line. For the
overlay gas, the flow rate of the 4-gas mix is also recorded by
a digital mass flow meter and a bank of solenoid valves. The gas
mix is in this case created by pulsing up to 4 different gasses
(air, oxygen, nitrogen and carbon dioxide) into the biocontainer.

Liquid Dosing
The tubing management system, when fully inflated, holds all
bioreactor addition lines with sterile connectors directly over Electronic Records (21 CFR 11)
the pump heads of the hardware. The Allegro STR bioreactors
The Allegro STR bioreactor controller offers all the technical
are equipped with four integrated pumps for liquid dosing.
features that are required to run the system compatible with
These cover the flow ranges that are needed for the respective
21 CFR 11. An administrator can set up users and define user
cascades (base, glucose, antifoam, etc.). For the addition of
rights and password. The password safety complies with the
larger liquid volumes, e.g. during filling or inoculation, external
requirement in 21 CFR 11 for electronic signatures. The electronic
pumps can be controlled via analog signals. All pumps, integrated
records are saved within the system and options for backup to
and external, can be made part of a defined control cascade.
external systems are available. The controller also offers an
OPC interface to fully control the operations on the bioreactor
from an external SCADA system.

Single-Use Biocontainer
The biocontainer is made of the transparent, low extractable
Allegro film as all other Allegro single-use systems, and which
has been well characterized for extractables and leachables.
The biocontainers are 100% integrity tested during manufacture.
All ports and connections are labeled with colored number
tags, providing guidance on how to set up and connect the
biocontainer to the hardware. Each system is provided with
multiple liquid addition lines and all are fitted with Kleenpak®
sterile connectors or Kleenpak Presto genderless sterile
connectors as standard.

Sensor Configurations
uL
 iquid temperature within the biocontainer is measured using
a Pt100 temperature sensor, this is surface mounted on the
Allegro STR 50 bioreactor
u Internal pressure within the biocontainer is measured
using a single-use pressure sensor supplied with each
single-use system
uL
 evel sensors in the hardware control the door interlocks
and temperature control to ensure there is a minimum liquid Allegro STR 100 biocontainer A
 llegro STR 50 biocontainer
volume prior to heating starting with Kleenpak Presto sterile
uT
 ransmitters for conventional pH and DO sensors are connectors shown on the
integrated in the controller and redundant transmitters are single-use biocontainer
available on selected models

6
Biocontainer Packaging Customization
In many manufacturers’ current use of a single-use bioreactor Pall has the standard biocontainer equipped with all liquid
design, there is risk of damage to the bioreactor bag during and gas connections for the most typical processes. Most
transportation and installation. It is therefore very important that operations, can be set up with the standard biocontainer. All
the biocontainer is properly packaged and the interaction of the ports are fitted with sterile connections. Process liquids, including
operator with the equipment is well defined. Protection of the media and inoculation, can easily and safely be transfered into
single-use bioreactor and the process it was being used in the bioreactor.
were considered from the outset of the Allegro STR bioreactor
Pall can assist in customizing the bioreactor systems. This
development.
includes small changes on the biocontainer up to integrating
For each size of the Allegro STR biocontainers, Pall has designed the bioreactor with all surrounding unit operations into a fully
innovative specific protective packaging. Some components of the automated process.
packaging are integral to the single-use system and provide
support for the connectors, tubing and filters in the single-use
system, thus reducing the risk of damage to the film during
transport. On the larger systems, the packaging material provides
ergonomic handles and guides to easily move and position the
single-use systems during unpacking and installation.

Setting up a transfer from an Allegro STR 50 L bioreactor to an Allegro

STR 200 L bioreactor

Documentation and Certificates

Impeller and Sparger Assembly The Allegro STR bioreactors are delivered with complete
documentation that qualifies the system. All units are tested prior
The Allegro STR bioreactors provide mixing via a bottom
to shipment and witnessed Factory Acceptance Testing (FAT) is
mounted pitched blade ‘elephant ear’ impeller. This shape
available on request.
has been selected from many candidates for good performance
in mammalian cell culture. The three 45 degree angle blades The biocontainers are delivered with all the batch specific
promotes efficient axial and radial mixing, while supporting certificates and calibration information for the included
options for both upward and downward flow depending on single-use sensors, where applicable.
the application required.
Pall also can assist with hardware qualification, and standard
The impeller is directly connected to the motor, which allows Installation Qualification (IQ), Operational Qualification (OQ), FAT,
excellent power transfer of approximately 0.25 W/kg into the and Site Acceptance Test (SAT) test protocols are available.
liquid. A dynamic seal arrangement at the bottom of the
biocontainer assures leak-free operation up to
the claimed service life.
The ring sparger and the CO2 removal line are mounted directly
under the impeller. This supports the effective distribution of the
gas bubbles in the liquid and increases their residence time in
the bioreactor. The sparger lines are fitted with sterilizing grade
Pall Emflon II gas filters. This design facilitates further effective gas
bubble distribution from the gas to liquid interface, maximizing
mass transfer required to support cell culture requirements.

www.pall.com 7
Pall Quality Standards uM
 aterials of construction of all components in contact with
fluid are certified TSE/BSE risk materials free
Pall maintains a very stringent approach to quality of purchased
uA
 re supplied double bagged and irradiated at a minimum
and manufactured components. The Allegro STR bioreactor
dose of 25 KGy
systems are designed and built to well recognized industry
standards. The Allegro STR system hardware meets many
quality standards including:

uG
 ood Automated Manufacturing Practice (GAMP)
current version
u CFR21 Part 11 for electronic records
uE
 lectromagnetic Compatibility (EMC) to EN 61326-1: 2013
and FCC CFR 47 Part 15B: 2013
uC
 E Marking to European Directives 2014/30/EU (2004/108/
EC extant until April 2016) and 2014/35/EU (2006/95/EC
extant until April 2016)
uT
 he product is made in a manufacturing facility where the
Quality Management System is ISO9001: 2008 certified
A full list of quality standards is available on request, please
contact your local Pall cell culture specialist.
The Allegro STR single-use biocontainers quality standards include:
uT
 he product is made in a manufacturing facility where the Preventive Maintenance and Service Packages
Quality Management System is ISO13485 & ISO9001 certified
The Allegro STR single-use bioreactors are supported with
u1
 00% of Allegro STR biocontainers are checked for correct extensive maintenance and service packages. Pall has a global
components, configuration, welding standard and dimensional network of technicians and engineers available to assist with
correctness. A visual inspection confirms there are no installation, qualification, training, technical support, preventive
embedded particles in the film or any loose particles inside maintenance and on-site repairs. Contact Pall to arrange to
or outside of the biocontainer discuss any of these services.
u1
 00% of Allegro STR biocontainers and seal housings
Pall Accelerator Process Development Services provide
undergo a pressure leak test during manufacturing
customers with expert advice and support for the implementation
uS
 amples from each biocontainer manufacturing batch of fully integrated single-use processing solutions into new or
undergo a tensile test of the film weld area & a microscope existing processes, including process optimization, equipment
inspection of the weld area to ensure the quality of the selection, transfer of existing processes, scale-up and
welded seal troubleshooting advice. Details can be found in the Accelerator
uF
 luid Path Endotoxins: Periodically, rinse effluents from Process Development Services brochure (USD 3079).
representative samples of Allegro systems are tested Preventive maintenance packages are available to ensure the
for endotoxins in accordance with USP <85> Bacterial continuous reliable operation of the Allegro STR bioreactors,
Endotoxins Test using Limulus Amoebocyte Lysate (LAL) minimizing downtime. These include a full functional test,
reagent. Fluid path rinses meet the internal specification exchange of wear parts and calibration of all sensors.
of < 0.25 EU/mL
uF
 luid Path Cleanliness: Periodically, rinse effluents from
representative samples of Allegro systems are tested for
particulates. Fluid path rinses meet the current limits under
USP <788> Particulate Matter in Injections
u The materials of construction of the Allegro STR
biocontainer meet:
•U
 SP<88> Biological Reactivity Test In Vivo for Class VI -
50 °C Plastics
• USP<87> Biological Reactivity Tests In Vitro, Cytotoxicity
• USP<85> Bacterial Endotoxin Tests
• USP<661> Physico-chemical Testing for Plastics
• USP<788> Particulate Matter in Injections

8
Technical Data

Allegro STR 50 Allegro STR 200 Allegro STR 1000 Allegro STR 2000
Bioreactor Bioreactor Bioreactor Bioreactor
Part number STR50-JC2302 STR200-JC2303 STR1000-JC STR2000-JC
Operational system space 1330 x 1550 x 1490 mm 1620 x 1121 x 2122 mm 1600 x 1700 x 2600 mm 1770 x 2010 x 2930 mm
requirements (W x D x H)
Weight (empty) 240 kg 720 kg 930 kg 1320 kg
Working volume 10 – 50 L 60 – 200 L 300 – 1000 L 400 – 2000 L
Impeller speed 25 to 200 rpm 25 to 150 rpm 25 to 105 rpm 25 to 105 rpm
Temperature range 4 to 40 ºC 4 to 40 ºC 4 to 40 °C 4 to 40°C
pH control 2 – 12 pH 2 – 12 pH 2 – 12 pH 2 – 12 pH
DO control 0 – 200% air saturation 0 – 200% air saturation 0 – 200% air saturation 0 – 200% air saturation
Conventional probe ports 6 4 6 6
Conventional probe 2 x pH, 2 x DO 2 x pH, 2 x DO 2 x pH, 2 x DO 2 x pH, 2 x DO
transmitters
Single-use sensor ports 2 x pH, 2 x DO 1 x pH, 1 x DO 1 x pH, 1 x DO 1 x pH, 1 x DO

Single-use sensor 2 x pH, 2 x DO 1 x pH, 1 x DO 1 x pH, 1 x DO 1 x pH, 1 x DO
transmitters
Temperature sensor 1 x Pt100 1 x Pt100 1 x Pt100 1 x Pt100
Analogue inputs 4 2 4 4
Load cell range 0 – 60 kg 0 – 250 kg 0 – 1100 kg 0 – 2500 kg
Connections for 4 2 2 4
external balances
Internal pumps 4 4 4 4
Connections for 2 1 2 2
external pumps
Pump speed 0 – 408 rpm 0 – 100 rpm 2 x 0 – 100 rpm 2 x 0 – 100 rpm
2 x 0 – 480 rpm 2 x 0 – 480 rpm
NOTE:
2Available with 110 V version (STR50-JC110); Allegro STR 50 bioreactor system available with standard second set single-use sensor transmitters

(STR50-JC230-R-SU & STR50-JC110-R-SU)

3Allegro STR bioreactor system also available with 110 V version (Part number STR200-JC110)

Ordering Information
The Allegro STR bioreactors are available as standard models
and both the bioreactor hardware and consumables can be
customized. Please contact a Pall representative to find the
ideal solution for your application. Our teams are specialized
in upstream and downstream processing and will gladly help
find the right technology for any part of your manufacturing
process. It is possible to combine the Allegro products to
meet any requirement in the upstream part of the process:

u Media preparation
u Buffer preparation
u Media sterilization and aseptic transfer
uL
 iquid transfer into the bioreactor (e.g. glucose, anti-foam,
base, etc.)
u Seed train solutions
u Cell harvest and separation
u Perfusion modules
Please contact Pall for a Total Solution discussion on your process.
www.pall.com 9
 Visit us on the Web at www.pall.com/biotech
E-mail us at biotech@pall.com
Corporate Headquarters International Offices
Port Washington, NY, USA Pall Corporation has offices and plants throughout the world in: Argentina, Australia, Austria, Belgium, Brazil,
+1.800.717.7255 toll free (USA) Canada, China, France, Germany, India, Indonesia, Ireland, Italy, Japan, Korea, Malaysia, New Zealand, Norway,
+1.516.484.5400 phone Philippines, Poland, Russia, Singapore, South Africa, Spain, Sweden, Switzerland, Taiwan, Thailand, United
Kingdom, and Vietnam. Distributors in all major industrial areas of the world. To locate the Pall office or distributor
biotech@pall.com e-mail nearest you, visit www.pall.com/contact.

European Headquarters The information provided in this literature was reviewed for accuracy at the time of publication. Product data
may be subject to change without notice. For current information consult your local Pall distributor or contact
Fribourg, Switzerland Pall directly.
+41 (0)26 350 53 00 phone
biotech@pall.com e-mail

Asia-Pacific Headquarters
© 2018, Pall Corporation. Pall, , Allegro, Emflon and Kleenpak are trademarks of Pall Corporation. ®
Singapore indicates a trademark registered in the USA. TM indicates a common law trademark. Filtration.Separation.
+65 6389 6500 phone Solution. is a service mark of Pall Corporation. uSiemens is a trademark of Siemens AG
sgcustomerservice@pall.com e-mail

10 4/18, PDF, GN18.07067 USD3117c