HyPerforma 5:1 Single-Use

Bioreactor (S.U.B.) User’s Guide

DOC0022 • Revision H
February 2021
Contents
Warnings, safety, and warranty information 1
How to use this guide 7

Chapter 1 HyPerforma Single‑Use Bioreactor (S.U.B.)

overview11
1.1 Introduction to the S.U.B. 12
1.2 Hardware characteristics 16
1.2.1 S.U.B. hardware components 16
1.2.2 S.U.B. system features 18
1.2.3 Additional system components 19
1.3 End user and third-party supplied components 26
1.3.1 pH and DO probes 26
1.3.2 Controllers 27
1.4 BPC characteristics 28
1.4.1 S.U.B. BPC components 28
1.4.2 S.U.B. BPC features 29

Chapter 2 Hardware assembly and setup 31

2.1 Initial installation preparation 32
2.1.1 Hardware shipment and setup 32
2.1.2 Hardware uncrating 32
2.1.3 Site preparation 32
2.2 Installation and setup 34
2.2.1 Preparing load cells 34
2.2.2 Leveling and connecting the system 35
2.2.3 Verifying drive shaft segments for 2,000 L systems 38
2.2.4 Setting the air pressure rate for motor lift adjustment
(1,000 and 2,000 L systems only) 38
2.2.5 Using the handheld controller for motor lift (1,000 and
2,000 L systems only) 39

Chapter 3 Operating information 41

3.1 General system operating information 42
3.1.1 BPC preparation 42
3.1.2 BPC handling instructions 42
3.1.3 BPC operating information 42
Contents

3.1.4 Hardware operating information 44

3.1.5 External data logging and control 47
3.2 BPC and drive shaft loading instructions for 50, 100,
and 250 L systems 47
3.2.1 Initial BPC loading steps for 50, 100, and 250 L
systems 47
3.2.2 Drive shaft insertion for 50, 100, and 250 L
systems 53
3.2.3 Final installation steps for 50, 100, and 250 L
systems 57
3.3 BPC and drive shaft loading instructions for 500
and 1,000 L systems 60
3.3.1 Initial BPC loading steps for 500 and 1,000 L
systems 60
3.3.2 Drive shaft insertion for 500 and 1,000 L systems 66
3.3.3 Final installation steps for 500 and 1,000 L systems 71
3.4 BPC and drive shaft loading, and condenser system
setup instructions for 2,000 L systems 74
3.4.1 Initial BPC loading steps for 2,000 L systems  74
3.4.2 Condenser system setup for 2,000 L systems 82
3.4.3 Drive shaft insertion for 2,000 L systems 94
3.4.4 Final installation steps for 2,000 L systems 100
3.5 Probe preparation and insertion 103
3.5.1 Preparation and sterilization 103
3.5.2 Making CPC AseptiQuik connections 105
3.5.3 Probe insertion 107
3.5.4 Probe calibration 109
3.6 Cell culture operating instructions 110
3.6.1 Operating conditions for cell culture applications 110
3.6.2 Checkpoints prior to media fill 111
3.6.3 Media fill 111
3.6.4 Agitation for units with electrical control panels 112
3.6.5 Agitation rate calculations 114
3.6.6 Drive shaft rotation 119
3.6.7 Temperature control 119
3.6.8 Sparging strategy 120
3.6.9 pH probe calibration 122
Contents

3.6.10 DO probe calibration 122

3.6.11 Checkpoints prior to inoculation 123
3.6.12 Cell inoculation 123
3.6.13 Volume scale up 124
3.6.14 In-process checkpoints 124
3.6.15 BPC sampling 124
3.6.16 Dispense and harvest 127
3.6.17 BPC disposal 128
3.6.18 S.U.B. shutdown 128
3.6.19 Preparation for the next run 129
3.7 Verification procedures 129
3.7.1 Mixing speed verification 129
3.7.2 Temperature controller verification 129
3.7.3 Pressure monitor verification (when present) 129
3.7.4 Load cell verification 130

Chapter 4 System features and specifications 131

4.1 Hardware features 132
4.1.1 Design features for 50–250 L systems 132
4.1.2 Design features for 500 L systems 133
4.1.3 Design features for 1,000 L systems 134
4.1.4 Design features for 2,000 L systems 135
4.2 Hardware specifications 136
4.3 E-Box features 154
4.4 BPC specifications 155
4.5 Additional system component part numbers 167

Chapter 5 Maintenance and troubleshooting 171

5.1 Maintenance 172
5.1.1 Routine maintenance 172
5.1.2 Preventive maintenance 172
5.2 Troubleshooting and frequently asked questions 174
5.2.1 Hardware operation issues 174
5.2.2 Cell culture operation issues 176
5.2.3 Sparging issues 177
5.2.4 Probe and connector issues 178
5.2.5 Other issues 179
Contents

Chapter 6 General ordering information 181

6.1 Ordering instructions 182
6.2 Ordering/support contact information 182
6.3 Technical support 183

Appendices Appendix A—Installation of female electrical receptacle

for units with AC motors and electrical control panels 184

Appendix B—Mettler Toledo IND331 display load cell

calibration instructions 187

Appendix C—2,000 L S.U.B. agitator operation and

maintenance guidelines 189

Appendix D—Drive shaft use log 190

Warnings, safety, and warranty information

Warnings, safety, and warranty

information
Thank you for purchasing this high-quality Thermo Scientific
equipment. We have included safety information in this guide, based
on our knowledge and experience. It is important, however, for you
to work with your Safety Management personnel to ensure that this
equipment is integrated into your safety practices. Please take some
time to perform your own job safety analysis in order to identify and
control each potential hazard.

WARNING: Read and understand this user's guide before

operating the equipment.
™ ™
The Thermo Scientific HyPerforma Single-Use Bioreactor (S.U.B.)
is designed to be operated under traditional eukaryotic cell culture
conditions. A general understanding of bioreactor systems and their
operation is important prior to using the system for the first time. Read
and understand this user’s guide before operating; failure to do so
could result in injury and potential loss of product.

WARNING: Hazardous voltage inside.

The mixer motor, motor controller and control panel all have
electrical components. There is a risk of electrical shock and
injury. Disconnect power before opening electrical components.
Service should be performed by certified personnel only.
Thermo Fisher Scientific recommends using standard lockout
procedures when working on electrical components. The main breaker
on the electrical control panel (E-Box) may be locked out.

WARNING: Static electricity may build up in BPCs.

• BioProcess Containers (BPCs) may act as insulators for
electrostatic charge. If electrostatic charge is transferred to a BPC,
the charge may be stored in the BPC and/or the product inside.
This phenomena varies by product and use; therefore, it is the sole
responsibility of the end user to ensure a hazard assessment is
conducted and the risk of electrostatic shock is eliminated.
• Where applicable, a product contact stainless steel coupler may be
grounded to the frame to dissipate electrostatic build up from the
material within a BPC. It is good practice to dissipate electrostatic
buildup by grounding all BPCs prior to coming in contact with them.
When working with BPCs, the use of nonconductive materials, such
as nonconductive gloves, is recommended.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 1

Warnings, safety, and warranty information

WARNING: Rotating parts—entanglement hazard.

Rotating and moving parts, such as the motor mount for 1,000 L and
2,000 L systems, can cause injury. Keep hands away from moving
parts during operation.
• Do not operate this equipment unless the supplied guarding is in
place and properly functioning.
• It is the responsibility of the end user to assess this equipment
and ensure that equipment and safeguards are in good working
condition, and that all operators are trained and aware of
entanglement hazards and associated protective devices, such as
hazard signs and guarding.

WARNING: Use ladders and elevated platforms with caution.

A few operations, such as loading a BPC into a large S.U.B., may
require the use of a ladder or platform. Before use, ensure the ladder
has been inspected and weight-rated for its user. When using a ladder
or platform, be sure it is stable, maintain three points of contact, and
make sure the steps are clean.

WARNING: Follow lockout/tagout procedures.

To prevent injury, when servicing equipment, use your company's
lockout/tagout procedures to isolate electrical, mechanical, pneumatic,
hydraulic, chemical, thermal, gravitational, or any other potential energy
and protect workers from the release of hazardous energy.

WARNING: Use caution with hazardous chemicals or materials.

Personnel servicing equipment need to know the hazards of any
chemicals or materials that may be present on or in the equipment.
Use general hazard communication techniques such as Safety Data
Sheets, labels, and pictograms to communicate any hazards.

WARNING: Potential confined space.

Operators may enter larger S.U.B. systems. Evaluate this equipment
against your confined space standards and procedures.

WARNING: Burst hazard—air under pressure.

The S.U.B. BPC chamber is under slight pressure under normal
operating conditions. Normal passive venting prevents any excess of
pressure building up within the chamber. Chamber pressure and inlet
line pressure should be monitored for proper settings.
• Contents under pressure
• Do not exceed 0.03 bar (0.5 psi) BPC pressure
• Do not exceed 0.34 bar (5 psi) inlet pressure
• Ensure vent filter is properly positioned and working properly

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 2

Warnings, safety, and warranty information

WARNING: Hot surface. Do not touch.

The heating jacket is designed to heat the inner vessel wall. Under
normal operating conditions, the motor will generate heat and could
create hot surfaces.
• Hot surface inside
• Contact with surfaces may cause burns
• Do not touch while in operation

WARNING: Pinch hazard.

The motor lift on 1,000 L and 2,000 L S.U.B.s can be raised and
lowered using the handheld controller. Caution should be used when
changing the position of the motor to avoid pinching an operator or
causing damage to the equipment or the BPC.

WARNING: Tipping hazard. The vessel should only be moved by

pushing using the provided handles or at the mid-point of the
vessel.
If pulled or moved too quickly, the vessel can tip, potentially leading
to damage to equipment or injury to personnel. To reduce the risk
of tipping, the vessel should only be moved slowly over smooth, flat
surfaces by at least two qualified personnel. During movement, any
locking feet should be retracted, and casters should be in the unlocked
position. The vessel should not be moved by pulling of any kind.

WARNING: The Thermo Scientific HyPerforma Single-Use

Bioreactor may not be installed in a potentially explosive
atmosphere as set forth in the applicable EU ATEX Directive.
It is the responsibility of the end user to review and understand the
potential dangers listed in the ATEX 2014/34/EU guidelines.

Protective earth grounding

Protective earth grounding must be verified prior to plugging the

S.U.B. into any electrical outlet. Ensure the receptacle is properly earth
grounded.

Environmental conditions

• Operating: 17°C to 27°C; 20 to 80% relative humidity, non-

condensing
• Storage: –25°C to 65°C
• Installation category II (over voltage) in accordance with IEC 664
• Altitude Limit: 2,000 meters

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 3

Warnings, safety, and warranty information

Electrical connections

Power should be supplied by a non-GFCI 15 amp circuit. Ground

faults occur when current is leaking somewhere, in effect, electricity is
escaping to the ground. Electrocution can occur when the human
body serves as the path for the leakage to the ground. A ground
fault circuit interrupter (GFCI) senses the current flowing to the ground
and switches off the power (trips the GFCI) in a fraction of a second at
currents well below those that are considered dangerous. Due to the
sensitivity of GFCIs to electrical leakage (a few mA), it is recommended
that the S.U.B. NOT be plugged into a GFCI outlet.

Water jacket vessel information

S.U.B. hardware units with water jackets have been designed to be

operated with water as the heat transfer medium, with temperatures
not exceeding 50°C (122°F) under less than 1 MPa (150 psig) operating
pressure. For the utmost safety it is recommended that the S.U.B. be
operated at 75 psig or less.

Note: The S.U.B. BPC operating limits for temperature are 5 to 40°C.
The internal pressure should not exceed 0.03 bar (0.5 psi). The water
jacket is not required to be registered, inspected and stamped with the
Code U symbol per section U-1(c)2(f) of the ASME Boiler and Pressure
Vessel Code and/or European Pressure Equipment Directive (PED)
97/23/EC. Upon request, a Declaration of Conformity, PED Sound
Engineering Practices can be made available.

Use of agitation speed governors and safety interlocks

Agitation speed governors set up on the bioreactor controller are used

to limit the maximum mixing speed, according to pre-defined liquid
volumes. Safety interlocks, which stop agitation when the volume in a
S.U.B. drops below defined limits, and speed-based governors prevent
damage to the drive shaft in the bioreactor. Agitation speed governors
and safety interlocks typically prevent the hazardous conditions listed
below.

• Operating the motor at any speed while loading the drive shaft
• Operating the agitator when volumes are less than 20% of a
system’s working volume
• Operating the agitator above recommended speeds based on
qualified power input to volume (P/V) thresholds

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 4

Warnings, safety, and warranty information

The hazardous conditions above must be avoided in order to ensure

qualified reliability. Using safety interlocks and agitation speed
governors eliminates the chance of human error, which could reduce
system reliability. Both the amount of liquid in the vessel and the
amount of power applied to the impeller have an impact on the applied
deflection on the shaft. Excess deflection and/or mixer speed may
damage the drive shaft.

For more information about using P/V and safety interlocks in

2,000 L bioreactor systems, see section 3.6.5 of this publication.

Warranty information

Any warranties, if applicable, covering this equipment exclude: (a)

normal wear and tear; (b) accident, disaster or event of force majeure;
(c) your misuse, fault or negligence; (d) use of the equipment in a
manner for which it was not designed; (e) causes external to the
equipment such as, but not limited to, external puncturing, power
failure or electrical power surges; (f) improper storage and handling of
the equipment; (g) use of the equipment in combination with equipment
or software that we did not supply; (h) equipment sold to you as ‘used’
products; (i) contact with improperly used or unapproved chemicals
or samples; (j) installation, removal, use, maintenance, storage, or
handling in an improper, inadequate, or unapproved manner, such as,
but not limited to, failure to follow the documentation or instructions in
the deliverables or related to the equipment, operation outside of stated
environmental or other operational specifications, or operation with
unapproved software, materials or other products; (k) manufacture in
accordance with requirements you gave us; (l) installation of software
or interfacing or use of the equipment in combination with software
or products we have not approved; (m) use of the deliverables
or any documentation to support regulatory approvals; (n) the
performance, efficacy or compatibility of specified components; and
(o) the performance of custom equipment or products or specified
components or achievement of any results from the equipment,
specified components or services within ranges desired by you
even if those ranges are communicated to us and are described in
specifications, a quote, or a statement of work. ADDITIONALLY, ANY
INSTALLATION, MAINTENANCE, REPAIR, SERVICE, RELOCATION
OR ALTERATION TO OR OF, OR OTHER TAMPERING WITH, THE
EQUIPMENT PERFORMED BY ANY PERSON OR ENTITY OTHER
THAN US WITHOUT OUR PRIOR WRITTEN APPROVAL, OR ANY
USE OF REPLACEMENT PARTS WE HAVE NOT SUPPLIED, WILL
IMMEDIATELY VOID AND CANCEL ALL WARRANTIES WITH
RESPECT TO THE AFFECTED EQUIPMENT. IF THE EQUIPMENT
IS TO BE USED IN THE UNITED STATES, WE MAY VOID YOUR
WARRANTY IF YOU SHIP THE EQUIPMENT OUTSIDE OF THE
UNITED STATES.
Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 5
Warnings, safety, and warranty information

Use restrictions

You must use this equipment in accordance with our documentation

and if applicable, with our other associated instructions, including
without limitation, a “research use only” product label or “limited use”
label license. This equipment is intended for research use or further
manufacturing in bioprocessing applications and not for diagnostic
use or direct administration into humans or animals, we do not submit
the equipment for regulatory review by any governmental body or
other organization, and we do not validate the equipment for clinical or
diagnostic use, for safety and effectiveness, or for any other specific
use or application.

Seismic guidance

The buyer of the equipment is responsible to ensure country specific

codes and seismic values are assessed for suitability of equipment
installation and safety at the designated site. In addition, it is the buyer’s
responsibility to assess the building structure for the designated
equipment to ensure correct seismic anchoring and tethering designs
for both the equipment and facility. It is highly recommended that
the buyer consult with a local, licensed third party architecture,
and engineering firm to provide the buyer with correct engineering
analysis and stamped documentation prior to equipment installation
at the facility. In addition, the buyer will be responsible for rigging and
anchoring of the equipment to a specified, fixed location. Thermo
Fisher can assist with establishing compliant seismic anchoring and
tethering designs for purchased equipment based on building and
country codes upon request at an agreed upon fee.

It is also noted that movable equipment (i.e. non-fixed or caster mount)

is exempt from seismic design requirements according to ASCE
7-16, Chapter 13, section 1.4. Although these units are exempt from
the seismic design requirements of ASCE 7, it should be noted that
such equipment is susceptible to overturning during a seismic event.
Therefore, it is the responsibility of the buyer to address seismic safety
for movable equipment at the designated facility.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 6

How to use this guide

How to use this guide

Scope of this publication

This user's guide contains information about the standard Thermo

™ ™
Scientific HyPerforma 5:1 S.U.B. systems, including hardware,
components, product design verification methods, installation,
operation, and specifications. It is intended for use by people who may
or may not have experience with Thermo Scientific systems, but who
have some knowledge of bioproduction processes and large-scale
mixing systems.

Document change information

Revision Date Section Change made Author
A 10/2016 -- Initial release S. Jelus/E. Hale
Warnings and Added pinch hazard and potential confined space warnings to
B 02/2017 E. Hale
Safety Warnings and Safety information
Warnings and Added information about safety interlocks to Warnings and Safety
B 02/2017 S. Jelus
Safety information
Added number information and photo of ends of mutiple-section
B 02/2017 2.2.4 S. Jelus/E. Hale
drive shaft
Added warning note about agitation rate and volume requirements,
B 02/2017 3.6.4 S. Jelus/E. Hale
and the use of safety interlocks
Added measurement to Table 5.1 for 2,000 L drive shafts and
B 02/2017 5.1.2 E. Hale
cross-reference to Appendix D
Added information about 2-piece drive shaft, a note about the
B 02/2017 3.4.3 position of impeller tubing inside the BPC, and serial number S. Jelus/E. Hale
information and a photo of ends of multiple-section drive shafts
Added information about and photo of the BPC bearing port cap
B 02/2017 1.1 E. Hale
label
Added a note about not pushing drive shaft straight into the
B 02/2017 3.4.3 S. Jelus/E. Hale
assembly when loading
Added information and Figure 3.102 to illustrate proper insertion of
B 02/2017 3.4.3 S. Jelus/E. Hale
drive shaft
Added 2,000 L BPC and drive shaft loading, and condenser system
B 02/2017 3.4 E. Hale
setup instructions
B 02/2017 4.2 Added 1,000 L and 2,000 L hardware specifications E. Hale
Added information about 2-piece drive shaft to 2,000 L
B 02/2017 4.2 S. Jelus/E. Hale
specifications
Added ceiling height requirements for 2-piece drive shaft and detail
B 02/2017 4.2 E. Hale
about mixing speed to 2,000 L specifications

B 02/2017 1.2.3 Added information about BPC lift for 2,000 L systems E. Hale
B 02/2017 4.3 Added drawing of 1,000 L electrical control panel E. Hale

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 7

How to use this guide

Document change information (continued)

Revision Date Section Change made Author

1.2.3 and Added information about motor lift and handheld controller for
B 02/2017 E. Hale
2.2.6 1,000–2,000 L systems
Added information about setting the air pressure rate for motor
B 02/2017 2.2.5 E. Hale
adjustment for 1,000–2,000 L systems
B 02/2017 3.6.5 Added Agitation Rate Calculations section to Chapter 3 E. Hale
B 02/2017 3.3 Added 1,000 L systems to 500 L BPC loading section E. Hale
B 02/2017 3.3.2 Added information about drive shaft serial numbers E. Hale
B 02/2017 1.2.1 Added drawing showing 2,000 L S.U.B. features E. Hale
B 02/2017 2.1.3 Added 1,000 L and 2,000 L electrical box preparation E. Hale
Added Appendix C—2,000 L S.U.B. agitator operation and S. Jelus/
B 02/2017 Appendix C
maintenance guidelines E. Hale
B 02/2017 Appendix D Added Appendix D—Drive shaft use log E. Hale
B 02/2017 Chapter 4 Removed 5:1 S.U.B. Made-to-order table E. Hale
B 03/2017 3.4.3 Added image of 2,000 L 5:1 BPC hanging tag E. Hale
B 03/2017 1.2.3 Added drive shaft specifications table E. Hale
B 04/2017 Warnings and Safety Added explosive atmosphere (ATEX) warning E. Hale
Changed "Maximum Mixing Rate" to "Agitation Speed Range" in
B 04/2017 4.2 E. Hale
hardware specifications
C 05/2017 Chapter 4 Corrected drive shaft diameter for 500 L S.U.B. E. Hale
Added harmonics/cavitation graph and context about these
D 09/2017 3.6 E. Hale
operating parameters to agitation information
Added a FAQ about excessive residue buildup in condenser bag
D 09/2017 5.2, 3.4 due to low TCU coolant levels, and provided more information in E. Hale
the 2,000 L condenser system instructions
Warnings, safety, and
D 09/2017 Added warranty and usage information E. Hale
warranty information
D 09/2017 How to use this guide Added a section for abbreviations/acronyms E. Hale
Added noise level to specifications for all S.U.B. sizes, and
D 09/2017 Chapter 4 corrected 2,000 L ceiling height requirement for 4-piece drive E. Hale
shaft loading
Added accessory part numbers, and BPC illustrations and
D 09/2017 Chapter 4 E. Hale
specifications
D 10/2017 3.4.3 Added note that all 2,000 L drive shaft heads are now black E. Hale
D 01/2018 -- Updated warning symbols and "Hot surface" warning E. Hale
D 01/2018 4.2 Corrected 500 L ceiling height requirement for drive shaft loading E. Hale
Revised Graph 3.4 by changing 2,000 L line to 750 and 1,000 L
D 01/2018 3.6.5 E. Hale
line to 375 L
3
D 01/2018 3.6.5 Changed Table 3.4 footnote to "> 20 W/m " E. Hale
3
D 01/2018 3.6.5 Changed first footnote in Table 3.6 to "> 20 W/m " E. Hale

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 8

How to use this guide

Document change information (continued)

Revision Date Section Change made Author

D 01/2018 5.1.2 Removed 2,000 L row from Table 5.1 E. Hale

Under "Drive shaft longevity and replacement," added "of cumulative

use" after "replacing your drive shaft every 360 days." In the second
D 01/2018 5.1.2 sentence of the second paragraph, changed to "every 180 days of E. Hale
cumulative use." In the first sentence of the note, added "< 50%
working volume."
D 01/2018 3.4.3 Updated Figure 3.102 to show the new deep impeller pocket E. Hale
Changed "Operating temperature range" to specifications for all
D 03/2018 Chapter 4 E. Hale
S.U.B. sizes to "Ambient to 40 ± 0.5°C (104 ± 0.9°C)"
D 04/2018 -- Removed references to 2,000 L S.U.B. four-piece drive shafts E. Hale
Warnings, safety,
D 08/2018 and warranty Added seismic guidance K. Leeman
information
Warnings, safety, Added emphasis to "Electrical connections" section, changed
E 11/2018 and warranty "certified personnel" to "Thermo Fisher Scientific service personnel," E. Hale
information and updated ATEX warning
How to use this Changed "Input into Thermo Scientific publications" section to
E 11/2018 E. Hale
guide "Questions about this publication"
E 11/2018 2.1.3, 3.6.4, 4.3 Updated text and images of the E-Box E. Hale
E 11/2018 3.5.3 Removed references to metal probe clips E. Hale
Removed Appendix B (AC-Tech variable speed drive settings) and
E 11/2018 Appendices E. Hale
renamed Appendices B through D, accordingly
Removed section 2.2.3 (Attaching the cable management system
E 11/2018 2.2.3, Various E. Hale
arm) and edited images showing the arm
E 12/2018 3.1.4, 3.6.4 Edited sentence (3.1.4), reworded step #2 (3.6.4) E. Hale
Updated accuracy in "Mixing speed verification" to ± 1.5 rpm or 1%
E 12/2018 3.7.1 E. Hale
of setpoint, whichever is greater
E 12/2018 4.2 Added tolerance to "Agitation speed range" in all specifications E. Hale
Minor formatting changes and updated cart length demension on
F 09/2019 4.2, Various T. Golightly
Figure 4.10
G 06/2020 -- Minor formatting revisions T. Golightly
Warnings, safety,
G 06/2020 and warranty Added Warning for Tipping Hazard T.Golightly
information
G 06/2020 3.2.1 Removed former Step 13, and removed former Figure 3.11 T. Golightly
G 06/2020 3.2.1 Updated image for Figure 3.10 T. Golightly
Added a CAUTION note below Step 12 for the BPC loading
G 06/2020 3.2.1 T. Golightly
instructions
G 06/2020 3.3.1, 3.4.1 Added a CAUTION note to the BPC loading instructions T. Golightly

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 9

How to use this guide

Document change information (continued)

Revision Date Section Change made Author

Corrected the overall width, length, and height in
H 10/2020 4.2 T. Golightly
Tables 4.2 and 4.4
H 10/2020 4.2 Replaced Figures 4.7–4.10 with updated dimensions T. Golightly
Updated the "Finesse pH and DO" sensors to
H 10/2020 1.3.1 T. Golightly
"Hamilton pH and DO" sensors in Table 1.2

Questions about this publication

If you have any questions or concerns about the content of

this publication, please contact technicaldocumentation@
thermofisher.com and your Thermo Fisher Scientific sales team.

Related publications
Please contact your local sales representative for information about the
related publications listed below.

Publication Description
Thermo Scientific HyPerforma 5:1 S.U.B. Validation Information about validation
Guide (DOC0023) procedures
Thermo Scientific HyPerforma 5:1 S.U.B. Data Sheets Product descriptions and ordering
(for various sizes) information
Thermo Scientific HyPerforma 5:1 S.U.B. Packing and Instructions for packing and
Unpacking Guide (DOC0033) unpacking equipment

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 10

How to use this guide

Abbreviations/acronyms

Refer to the list below for definitions of the abbrieviations and acronyms
used in this publication.

BPC BioProcess Container

cGMP Current good manufacturing practices
DO Dissolved oxygen
ETP Equipment Turnover Package
GFCI Ground fault circuit interrupter
HMI Human machine interface
ID Inner diameter
IEC International Electrical Code
OD Outer diameter
PED Pressure Equipment Directive
PID Proportional integral derivative
P/V Power input to volume
RTD Resistance temperature detector
STR Stirred tank reactor
S.U.B. Single-Use Bioreactor
TCU Temperature control unit
VFD Variable frequency drive

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 11

 HyPerforma Single‑Use
Bioreactor (S.U.B.)
1
overview

Chapter contents
1.1 Introduction to the S.U.B.
1.2 Hardware characteristics
1.3 End user and third-party supplied components
1.4 BPC characteristics

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 12

Chapter 1 | S.U.B. overview

1.1 Introduction to the S.U.B.

The Thermo Scientific HyPerforma Single-Use Bioreactor (S.U.B.) has
been designed as a single-use alternative to conventional stirred tank
bioreactors currently utilized in eukaryotic cell culture. Based on years
of accepted stirred tank reactor (STR) design, the S.U.B. emulates STR
scalability and operating parameters, yet it has the unique advantage
of being a single-use device. Ease of setup with respect to system
operation, and integration into existing facilities makes the S.U.B. an
attractive alternative to its conventional STR counterpart.

The 5:1 S.U.B. allows users to take advantage of low volume mixing
in addition to half and full volume, providing greater flexibility all in
one system. Critical design parameters such as height-to-diameter
ratios, mixer design and location, and typical control system interfaces
have been maintained. A key element to the single-use design is the
plastic (polyethylene) impeller with a bearing/seal assembly linking to
an external mixer drive. Quick setup and changeover allows for faster
turnover in cell culture runs over traditional reusable systems.

The S.U.B. system consists of the following primary components:

1. Outer support container with water jacket heating system
2. S.U.B. BioProcess Container (BPC) that is supplied gamma
irradiated
3. Control system for units with AC motors for agitation
4. Direct drive agitation mixing assembly with an AC or DC motor
(DC motor not available for 1,000 and 2,000 L systems), drive shaft,
and impeller

Figure 1.1. 50–500 L S.U.B.s.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 13

Chapter 1 | S.U.B. overview

The outer support container is engineered and fabricated to fully

support each BPC and allow easy access for operation. It is a stainless
steel vessel that holds and supports the BPC. The outer support
container contains the mixing drive and water jacketed tank on casters
(2,000 L S.U.B.s are not on casters). The drive shaft is detachable and
reusable, and is inserted into the BPC through the mixing assembly
and into the bearing port. Load cells are standard for all 5:1 S.U.B.
systems.

The BPC includes the impeller assembly, sparger, vent filter inlet/
outlet ports, probe integration ports, filling, dispensing, and sampling
ports. Each BPC comes fully assembled and gamma irradiated. The
materials are fully qualified for biological product contact per USP
Class VI plastics. Each assembly is manufactured under current Good
Manufacturing Practices (cGMP) and is supported by qualification
and validation information. Innovative, proprietary technology allows
for the integration of the mixing shaft and pH/dissolved oxygen (DO)
probes, and the resistance temperature detector (RTD). The probe and
temperature interfaces are comparable to traditional systems with the
design allowing for simple aseptic connections. Integrated spargers
(drilled hole, overlay, and cross flow) are built into the BPC through
universal ports.

The Thermo Scientific S.U.B. utilizes an open architecture design for

the control system, allowing for integration with customer systems
or with third-party controllers for feed pumps, mass flow controls,
and human-machine interface (HMI) screens. Controls for agitation
are integrated into the S.U.B., with pH/DO probes and controls being
supplied by the user or a third-party integrator. HyPerforma S.U.B.
systems require a temperature control unit (TCU) selected and supplied
by the end user or by Thermo Fisher Scientific.

Figure 1.2. 2,000 L 5:1 S.U.B.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 14
Chapter 1 | S.U.B. overview

This user’s guide covers the setup, operation, maintenance, and

troubleshooting of all 5:1 S.U.B. systems in the following volumes: 50,
100, 250, 500, 1,000, and 2,000 L.

Figures 1.3–1.7 illustrate some of the standard labeling on 50,

100, 250, and 500 L 5:1 S.U.B. systems. Systems in these sizes
include the following:
• Hanging tag labeled for "5:1 Mixing" attached to the BPC bearing
port (Figures 1.3 and 1.4)
• Motor mount engraved with "5:1 Mixing" (Figures 1.5 and 1.6)
• "Usable by 5:1 drive shaft only" warning label on BPC bearing port
cap (Figure 1.7)
• Drive shaft with a black head that is engraved with "5:1 Mixing."

Figure 1.4. Close-

Figure 1.3. Hanging tag on 50–500 L 5:1 BPC bearing port.
up view of 5:1 BPC
hanging tag.

Figure 1.5. Engraving on 50–500 L 5:1 motor mount. Figure 1.6. Close-up
view of engraving on
5:1 motor mount.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 15

Chapter 1 | S.U.B. overview

Figure 1.7. Label on 50–500 L 5:1 BPC

bearing port cap.

Important notes:
• If your 50, 100, 250, or 500 L system components are not labeled
for 5:1 mixing, consult your sales representative immediately.
• There are no 5:1 mixing labels for 1,000 and 2,000 L S.U.B.s and
BPCs.
• 5:1 S.U.B. mixing speeds should be reduced when operating at
lower volumes to avoid damaging the system. For recommended
agitation rates, see section 3.6.4 and the hardware specifications in
section 4.2 of this user's guide.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 16

Chapter 1 | S.U.B. overview

1.2 Hardware characteristics

1.2.1 S.U.B. hardware components

Figures 1.8 and 1.9 below illustrate all available components of a

500 L S.U.B. system. Note: 50, 100, and 250 L systems do not have a
BPC loading door, and use a one-piece drive shaft.

4 11

2
5
12
3

13
6

15
7 14
16
18
8
10

17 19
9
Figure 1.8. Front view of 500 L S.U.B. Figure 1.9. Back view of 500 L S.U.B.

1. Exhaust vent filter holder 11. Standard tool set: 10 mm (3/8 in.) x 16.9 Nm (150
2. Mixing assembly with shield in-lb.) square torque wrench, load cell and motor cap
3. Mixer motor lockout wrench
4. Bearing port receiver with clamp 12. Cable management system
5. BPC loading door (500 L only) and liquid sight 13. Drive shaft, stored (2-piece)
windows 14. 304 Stainless steel outer support container
6. Electrical control panel (E-Box), optional 15. 0.95 cm (3/8 in.) Dimpled jacket
7. Probe hanger bracket 16. Bleed valve
8. Probe access windows 17. Cart assembly
9. Leveling casters 18. Bottom cutouts/pins for BPC attachment/alignment
10. Load cells 19. Quick-connect water inlet/outlet ports

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 17

Chapter 1 | S.U.B. overview

Figures 1.10 and 1.11 below illustrate all available components of a

2,000 L S.U.B. system. Note: 1,000 L systems have a cutout instead of
a back access door, and do not include a BPC lift. See section 4.1.3 for
a complete illustration of a 1,000 L S.U.B.

1 10

2 11

11
3

4 19
15
12

7 16

13 20
8

9 14

21

17
22

18
Figure 1.10. Front/side view of 2,000 L S.U.B. Figure 1.11. Back view of 2,000 L S.U.B.

1. Motor lift 13. Load cell display

2. Mixer motor 14. Electrical control panel (E-Box), optional
3. Mixing assembly with motor safety shield 15. 0.95 cm (3/8 in.) Dimpled jacket
4. Handheld controller for motor adjustment (on bracket) 16. BPC lift control
5. Emergency stop (E-Stop) 17. Bleed valve
6. Drive shaft, stored 18. Quick-connect water inlet/outlet ports
7. BPC loading door and liquid sight windows 19. 304 Stainless steel outer support container
8. Probe access window 20. Rear access door
9. Probe hanger bracket (with probe clips) 21. Bottom cutouts/pins for BPC attachment and alignment
10. Exhaust vent filter holder 22. Load cell (3)
11. BPC lift, for 2,000 L 5:1 S.U.B.s only (2)
12. Standard tool set: 10 mm (3/8 in.) x 16.9 Nm (150 in-lb.)
square torque wrench, load cell and motor cap lockout wrench

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 18

Chapter 1 | S.U.B. overview

1.2.2 S.U.B. system features

The S.U.B. is designed for system mobility and easy integration, and
utilizes a straightforward operator interface. The following sections give
general descriptions of S.U.B. hardware features.

Agitation
If your system uses an AC motor and a Thermo Scientific electrical
control panel (E-Box), the stirring speed is adjusted by using the E-Box
keypad interface. The agitation control interface utilizes a digital display
to indicate stirring speed in units of revolutions per minute (rpm). Power
is supplied to the motor by a two-position power switch. The up and
down arrows on the agitation keypad adjust the stirring speed. If
your 50, 100, 250, or 500 L system has a DC motor and is integrated
and managed by a third-party controller, agitation is managed by the
controller. Thermo Fisher Scientific does not provide electrical control
for units with DC motors.

Bioreactor control system

The S.U.B. is designed to integrate with existing bioreactor control
systems in their numerous configurations. The S.U.B. control system
supplied with the Thermo Scientific E-Box manages the agitation
process parameters. Parameters of pH and DO, gas management,
feed addition, and base addition control must be managed by an
external controller supplied by the end user or a third-party integrator.

Temperature
The S.U.B. can be operated within the temperature range from ambient
to 40°C. The process temperature is measured by means of a supplied
RTD (pt-100) that is inserted into the thermowell of the BPC. Water
jacket system temperature control is maintained through the TCU.

Condenser system
The condenser system is recommended for 2,000 L S.U.B. use, and
is available as optional hardware for smaller systems. It cools exhaust
gases and re-circulates condensate into the bioreactor.

Heating performance
Heating times for the S.U.B. systems vary based upon operating
liquid volume and temperature, ambient or heating fluid temperature,
sparger rate, and mixing rate. Users should adjust process liquid
staging and seeding strategies to the unique aspects of the S.U.B.
Process controllers are designed to provide optimum heat transfer
and to minimize heat-up times while maintaining the material integrity
of the polymer film construction of the BPC. Refer to section 3.1.4 for
expected heating times.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 19

Chapter 1 | S.U.B. overview

1.2.3 Additional system components

Drive shafts
The drive shaft couples the mixing impeller that is inside the BPC to
the motor. If you are using a 5:1 BPC, it is important to use the correct
drive shaft. See Table 1.1 for drive shaft specifications for each S.U.B.
size.

Table 1.1. Drive shaft specifications for 50–2,000 L systems.

System Overall shaft Number of
Head description Material
size length pieces
Black, engraved with Stainless steel
50 L 85.09 cm (33.5 in.) 1-piece
"5:1 Mixing" or aluminum
Black, engraved with Stainless steel
100 L 100.58 cm (39.6 in.) 1-piece
"5:1 Mixing" or aluminum
Black, engraved with Stainless steel
250 L 120.90 cm (47.6 in.) 1-piece
"5:1 Mixing" or aluminum
Black, engraved with Stainless steel
500 L 155.7 cm (61.3 in.) 2-piece
"5:1 Mixing" or aluminum
3-piece or
1,000 L 167.6 cm (66 in.) White Stainless steel
4-piece
Black, engraved with
2,000 L 215.6 cm (84.9 in.) 2-piece Carbon fiber
drive shaft length

Probe integration
The autoclave tray (Figure 1.12) holds the electrochemical probes and
bellows in place during the autoclave sterilization process. Design
elements include the following.
• Fabricated from stainless steel
• Features a plastic handle for easy transport right out of the
autoclave
• Positions probes on 15% incline for greater probe/membrane
longevity
• Will restrain probe bellows from collapsing during sterilization
• Accommodates two probes

Handle

Probe assembly

Autoclave tray
for probe kits

Figure 1.12. Autoclave tray and probe assembly.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 20

Chapter 1 | S.U.B. overview

The probe assembly (Figure 1.13) is an innovative design to package

user-supplied pH and DO probes for sterilization, and to aseptically
connect them to the BPC. The probe assembly includes an aseptic
connector, molded bellows cover, and threaded probe adapter.

Molded bellows cover

Threaded probe
Aseptic adapter
connector

Figure 1.13. Probe assembly.

BPC lift (2,000 L systems only)

The BPC lift (Figure 1.14) helps users load the BPC into 2,000 L
S.U.B.s. Two hooks, on opposite sides of the S.U.B., attach to tabs on
the BPC. The BPC lift control (Figure 1.15) is used to raise and lower
the BPC. For instructions on using the BPC lift, see section 3.4 of this
publication.

Figure 1.14. BPC lift for 2,000 L systems.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 21

Chapter 1 | S.U.B. overview

Emergency stop
(E-Stop)

Motor enable switch

Vertical lift
switch

Main
disconnect
switch

Figure 1.15. BPC lift control for 2,000 L

systems.

Motor lift and handheld controller (1,000 and 2,000 L systems

only)
1,000 and 2,000 L 5:1 S.U.B. systems include a motor lift (Figure 1.16),
used to adjust the position of the motor. The motor lift is operated
by a handheld controller (Figure 1.17), which is located on a bracket
attached to the front of the S.U.B. For instructions on how to operate
the handheld controller, see section 2.2.5 of this guide—Using the
Handheld Controller for Motor Lifts (1,000 and 2,000 L systems only).

Figure 1.16. Motor lift for 1,000–2,000 L systems. Figure 1.17. Handheld controller
for motor lifts.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 22

Chapter 1 | S.U.B. overview

Options and accessories

The following additional system components may or may not be
installed on your S.U.B. system. To order accessories for retro-fitting to
your unit, contact your sales representative.

Exhaust vent filter heaters

The exhaust vent filter heater system, which includes the heater, a
controller, and power cord (Figure 1.18), is available for increased
longevity of the exhaust filter on the BPC. The heating element is fully
insulated with molded silicone and secured around the filter by use
of snap retainers, fully encapsulating the exhaust filters for consistent
temperature regulation. Heating the filter sufficiently to eliminate
the formation of condensation reduces the risk of fouling the filter
membrane.

The heater is factory preset to operate between 40°C–50°C, but can

easily be adjusted to the demand of the application. Temperature
settings above 60°C are not recommended.

Figure 1.18. Vent filter heater.

Load cells
Load cells, which are used to determine the weight of the contents of a
S.U.B., are installed on all standard 5:1 S.U.B. systems. Load cell retro-
fit kits can also be added to existing S.U.B. units by a certified service
technician. Load cells arrive uncalibrated. The load cell manufacturer or
a qualified technician should calibrate these systems onsite.

The load cell kit comes with three load cells, summing block,
wiring, and a display screen with a choice of several data interfaces
(Figure 1.19).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 23

Chapter 1 | S.U.B. overview

Figure 1.19. Load cell system overview.

Load cells are typically radial-mounted in sets of three. The mounting

location (Figure 1.20) varies slightly for each size in order to allow easy
access to the bottom drain or sparging mechanisms and tubing.

Figure 1.20. Load cell location.

AC and DC motors
AC and DC motor options are available to help tailor the S.U.B. system
to your specific needs. Note: DC motors are only available for 50, 100,
250, or 500 L systems.

DC motors operate at a lower voltage and, when integrated with a

controller system that receives sensor feedback, provide more accurate
speed control through a digital program transmitter. The DC motor
comes with an encoder, but does not come with a motor control
option from Thermo Scientific. For DC motors, a control option must be
specified by the end user.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 24

Chapter 1 | S.U.B. overview

AC motors may be used with the Thermo Scientific E-Box. AC motors

include a variable frequency drive, and are controlled using either the
provided keypad or a controller specified by the end user.

Cable management systems

The cable management system is available as an option on 50, 100,
250, 500, and 1,000 L units. It is used to organize various lines and
includes the following components (Figure 1.21).

• Internal channel for sparge lines

• External channels for feed and base addition lines
• Harvest line hook

Channels for
sparge lines

Channels for
feed and base
addition lines

Harvest line
hook

Figure 1.21. 500 L S.U.B. with

cable management system.

Miscellaneous items
The miscellaneous items listed below are ancillary components
that support the operation of the HyPerforma S.U.B. for cell culture
production, and enhance the overall performance of the complete
system.
• Sampling manifold with luer lock
• S.U.B. temperature sample port—For RTD calibration/validation

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 25

Chapter 1 | S.U.B. overview

• Sparge line support—Keeps the drilled hole sparge line in

a vertical position for optimal gas flow (Figure 1.22). For more
information see section 2.2—Installation and Setup.

Figure 1.22. Sparge line support.

• Heavy-duty tubing clamps (typically four or five)—Tubing

clamps (Figure 1.23) are required for pinching off line sets that are
not in use in order to prevent process fluids from moving into the
line sets. Prior to sterile probe insertion, tubing clamps must be in
place to close off probe ports. For more information, see the BPC
and drive shaft loading instructions in sections 3.2, 3.3, and 3.4 of
this publication.

Figure 1.23. Heavy-duty tubing clamps.

Note: The sparge line support is included with all standard S.U.B.
units. Other items are sold separately. Please contact your sales
representative for more information.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 26

Chapter 1 | S.U.B. overview

1.3 End user and third-party supplied

components

1.3.1 pH and DO probes

Table 1.2 shows the length and diameter requirements for traditional
sensors (probes) that can be integrated into the S.U.B. These
requirements are based on the necessary insertion depth of the probe
when used with the probe ports. Note: The presence of a properly
positioned O-ring on the probe is critical for use with the S.U.B.

Table 1.2. Manufacturers and models of compatible pH/DO probes.

O-ring to probe tip
Probe lengths (from O-ring to tip) must not exceed 235 mm
Print/lit. Actual
Probe Part number Diameter Thread type Length Length
12 mm 235 mm 235 mm
AppliSens DO Z010023525 13.5 PG
(0.47 in.) (9.25 in.) (9.25 in.)
12 mm 235 mm 235 mm
AppliSens pH Z001023551 13.5 PG
(0.47 in.) (9.25 in.) (9.25 in.)
InPRO 6800/12/220, PN 12 mm 215 mm 215 mm
Mettler Toledo DO 13.5 PG
52200966 (0.47 in.) (8.46 in.) (8.46 in.)
405-DPAS-SC-K8S/225, PN 12 mm 195 mm 219 mm
Mettler Toledo pH 13.5 PG
104054481IG (0.47 in.) (7.67 in.) (8.62 in.)
12 mm 215 mm 214 mm
Broadley-James DO D140-B220-PT-D9 13.5 PG
(0.47 in.) (8.46 in.) (8.42 in.)
12 mm 225 mm 219 mm
Broadley-James pH F-635-B225-DH 13.5 PG
(0.47 in.) (8.85 in.) (8.62 in.)
12 mm 225 mm 220 mm
Hamilton DO 237542 13.5 PG
(0.47 in.) (8.85 in.) (8.66 in.)
12 mm 225 mm 220 mm
Hamilton pH 238633-2543 13.5 PG
(0.47 in.) (8.85 in.) (8.66 in.)
Note: Consult the probe manufacturer’s website for appropriate probe cable connection and part number.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 27

Chapter 1 | S.U.B. overview

1.3.2 Controllers

Thermo Scientific products are designed with an open-architecture

approach to the integration of controls. Our industry-leading S.U.B.
has been integrated with most controllers on the market, allowing
customers to choose the control system they want, or to reduce
expense by integrating with a controller that is already onsite. In order
to facilitate integration, electrical schematics are provided in the ETP
supplied with the HyPerforma S.U.B. Companies that offer control
solutions in either current Good Manufacturing Practices (cGMP) format
or non-cGMP format for Thermo Scientific S.U.B. units are listed below.

• ABEC
• Bellco
• Broadley-James
• Dasgip
• Emerson
• Honeywell
• New Brunswick Scientific
• Pendotech
• Sartorius Stedim Biotech

The HyPerforma 5:1 S.U.B is also available as a complete turnkey

system through Thermo Fisher Scientific. These S.U.B. units may be
provided with integrated controls, pump towers, a control monitor, and
advanced features such as data logging, multiple S.U.B. connections,
and optional 21CFR part 11 compliance for cGMP manufacturing. A
variety of single-use sensors are available for pH, DO, and pressure
control. Thermo Fisher Scientific can provide complete, integrated
solutions using the manufacturers listed below.

• Allen-Bradley
• Applikon PLC eZ-controller
• Emerson Delta V
• Finesse PC controller
• Siemens

Contact your local sales representative for more information.

Note: The S.U.B. will work well with any of the various control system
platforms, such as PLC, PC, DCS, or proprietary operating system
based controllers.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 28

Chapter 1 | S.U.B. overview

1.4 BPC characteristics

1.4.1 S.U.B. BPC components

Figure 1.24 illustrates the components of a standard 5:1 500 L

BPC. Some components vary between 50–250 L and 500–2,000 L
systems; see Table 1.3 on the following page for more information.
Note: 2,000 L BPCs may also have an optional condenser system.

1. Exhaust vent
filter (2)

2. Inoculum
addition port

3. Base addition 6. Gas overlay port

port

4. Media
addition port 7. Hanging tabs (8)

5. Feed port

8. Seal/bearing assembly

9. Impeller

10. Cross flow sparger

11. Ports with aseptic

connectors

12. Temperature RTD/

sampling port

14. Drilled hole 13. Drain port

sparger

Figure 1.24. Typical 500 L BPC.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 29

Chapter 1 | S.U.B. overview

Table 1.3. BPC information for Figure 1.24.

Component Description
Single-use capsule filter for exhaust gas exchange; 50, 100, and 250 L units
1. Exhaust vent filter
have one filter, and 500, 1,000, and 2,000 L units have two filters
2. Inoculum addition port For inoculum addition
3. Base addition port For base addition
4. Media addition port For addition of media
5. Feed port For addition of other liquids
6. Gas overlay port For the introduction of air or other gases; protected by gas filter
7. Hanging tabs (8) For securing BPC inside the S.U.B.
Links with the mixer motor and allows the impeller to turn while retaining
8. Seal/bearing assembly
integrity of the S.U.B. BPC
9. Impeller Injection molded plastic; Links to seal/bearing assembly by C-Flex tubing
10. Cross flow sparger Supplies oxygen during low-volume mixing
11. Ports with aseptic connectors For integration of standard 12 mm (0.47 in.) monitoring pH and DO probes
12. Temperature RTD/sampling For integration of the temperature probe while retaining integrity of the S.U.B.
port BPC/needleless sampling or connection to the sampling manifold
13. Drain port Used when draining the S.U.B.
For the introduction of air, oxygen or other gases; integrated into the chamber
14. Drilled hole sparger*
and protected by gas filters
*Note: 2,000 L BPCs have two drilled hole spargers.

1.4.2 S.U.B. BPC features

The cell culture itself will be contained inside the gamma irradiated
BPC. The chamber is manufactured from film, which is a co-extruded
structure specifically designed for biopharmaceutical process usage.
All materials are qualified for a broad range of physical, mechanical,
biological, and chemical compatibility requirements. Refer to data
in our BPC catalog and film validation guides; contact your sales
representative for a copy.

Spargers
The standard 5:1 BPC is designed with special spargers (drilled hole,
cross flow, and overlay) that produce very efficient mass transfer of
oxygen. They typically require much less gas inflow than conventional
spargers. Gas flow rates supplied as overlay or through the cross
flow sparger should also be reduced as much as possible; this will
minimize both liquid evaporation and demand on the exhaust filter.
Minimizing gas flow through the drilled hole sparger reduces the
occurrence of foam in the headspace that may plug the exhaust filter.
For more information, refer to section 3.1.3—Operating Pressure, and
section 3.6—Cell culture operating instructions, in this user’s guide.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 30

Chapter 1 | S.U.B. overview

Exhaust vent filter

The exhaust vent filter used on 50–1,000 L S.U.B.s is a Pall™ KA3
series filter utilizing hydrophobic PVDF membranes. To maintain a
sterile connection, the standard BPC is supplied with the filter arrow
pointing toward the BPC. This ensures that the filter vents are outside
of the sterile connection. For users with more demanding applications,
an optional vent filter heater can be used.

The exhaust vent filters used on 2,000 L S.U.B.s are Meissner™

UltraCap™ series filters utilizing hydrophobic PVDF membranes. These
filters are provided in normal orientation with the flow arrow on the filter
housing pointing away from the BPC. The normal orientation provides
maximum filter capacity. No side vents are provided. Condensate must
be managed by use of the condenser system or vent filter heater.

Connections
Multiple aseptic connection options exist for S.U.B. users. Standard
BPCs include tubing welder sections, quick-connects, and CPC™
™
AseptiQuik connections. The BPC is designed with various lengths
and dimensions of thermoplastic tubing for the purpose of adding to
and dispensing from the BPC.

Sampling port
The S.U.B. is equipped with a small volume sample port that is
adjacent to the BPC thermowell. This small-diameter silicone dip tube
of 152.4 mm length (6 in.) allows low void volume samples to be taken
for cell viability and density, as well as analyte analysis. This dip tube
is supplied with a luer lock connector (SmartSite™) that allows for
direct sampling or attachment of various sampling manifolds by use of
standard luer lock connection. Alternatively, manifolds can be welded
onto the C-Flex sample line using a tubing welder.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 31

 Hardware assembly
2
and setup

Chapter contents
2.1 Initial installation preparation
2.2 Installation and setup

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 32

Chapter 2 | Hardware assembly and setup

2.1 Initial installation preparation

2.1.1 Hardware shipment and setup

The Single-Use Bioreactor (S.U.B.) hardware will arrive crated. For

unpacking instructions and detailed contents of the crate, please refer
to the Thermo Scientific HyPerforma 5:1 S.U.B. Packing and Unpacking
Guide (DOC0033), and the packaging drawings, which are included
in the shipping crate. Be sure to follow the unpacking instructions
provided and retain all packaging materials.

2.1.2 Hardware uncrating

The S.U.B. hardware will arrive with the following items:

• Outer support container (platform, tank, and control panel)
• Drive shaft, resistance temperature detector (RTD), four probe
brackets, and standard tool set (spanner wrench and torque
wrench)
• Equipment Turnover Package (ETP), provided on a USB drive
(shipped separately)

Detailed instructions for crating, uncrating, and assembly of 50,

100, 250, 500, 1,000, and 2,000 L S.U.B. units are included in the
Thermo Scientific HyPerforma 5:1 S.U.B. Packing and Unpacking
Guide (DOC0033). After uncrating, contact your sales representative
immediately if any damage has occurred.

2.1.3 Site preparation

Electrical connections for units with AC motors and an

electrical control panel
S.U.B. hardware using AC motors cannot be used on circuits equipped
with ground fault circuit interrupter (GFCI) circuit protection because of
the potential for nuisance tripping. The electrical plug on the S.U.B. is
a connector that offers a secure ground. These connectors meet the
electrical safety codes for portable equipment and are International
Electrical Code (IEC) rated (meet IEC standard 60309). This plug
provides electrical ground prior to power connection. The supplied
electrical receptacle should be hardwired into the facility by a qualified
electrical technician; for U.S. installations, the receptacle will require the
use of an adapter mounting plate (supplied), which will fit into a two-
gang box. For additional information on the adapter mounting plate,
please see the ETP. Alternatively, the system can be hardwired directly
into the facility. Note: The yellow plug and receptacle are for 120 VAC,
and the blue are for 240 VAC S.U.B.s.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 33

Chapter 2 | Hardware assembly and setup

Electrical connections for 50, 100, 250, and 500 L systems with
DC motors
S.U.B. units using DC motors are not supplied with electrical control
panels (E-Boxes). When using a DC motor, electrical connections must
be supplied by a third-party integrator.

Outer support container preparation

Each outer support container is shipped directly from the manufacturer,
and arrives with various safety mechanisms in place. Follow the
guidelines below to set up the S.U.B. upon arrival.

WARNING: Any procedure that requires the E-Box to be opened

should be performed with the main electrical disconnect in the
locked out position and all power sources removed from the E-Box.
For operator safety, secure the location of the S.U.B. outer support
container by disabling the swivel casters before servicing.

Electrical preparation for 50–2,000 L systems with AC motors

and E-Boxes
1. Using a flat-head screwdriver, open the E-Box and locate the
breakers for the pressure sensor, continuous power outlets
non E-stoppable (2), and continuous power outlets E-stoppable
(2) (Figure 2.1). These breakers should be in the "on" position
during operation, which will be in the "up" position or pressed in,
depending on the breaker type. For electrical schematics, please
refer to the ETP, which is provided on a USB drive.

VFD breaker

Main power
breaker

Temp. display
breaker

Pressure
breaker

E-Stop power
breaker

Figure 2.1. 50–2,000 L S.U.B. E-Box interior. Continuous power

breaker

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 34

Chapter 2 | Hardware assembly and setup

2. Close the E-Box and lock the panel using a flat-head screwdriver
before continuing.

2.2 Installation and setup

2.2.1 Preparing load cells

All manual movements of mobile S.U.B. hardware should be over

smooth surfaces, with the S.U.B. empty and disconnected from all
power and gas/feed sources. All load cells must be fully locked down
in order to move the S.U.B.

Follow the steps below to prepare load cells for use. Figure 2.2
illustrates the location and components of load cells, which will be
referenced throughout the load cell preparation process.

Lockout
nut

38.1 mm
(1.5 in.)
Tri-clamp

Lockout
A
post

Delrin slip
ring
Figure 2.2. Close up view of load cells.

1. For S.U.B. hardware units purchased with factory-installed load

cells, the load cells are shipped in the locked position (threaded up)
for equipment protection.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 35

Chapter 2 | Hardware assembly and setup

2. To unlock the load cells, remove and discard the delrin slip ring
if it is present. Remove the tri-clamp. Loosen the lockout nut,
using the small end of the supplied tool (Figure 2.3), until the nut is
tight against the base or leg of the S.U.B. Repeat this process for
each load cell until all of the lockout nuts are disengaged from the
lockout posts. Do not reinstall the tri-clamp.

Drive shaft
cap end Load cell
lockout end

Figure 2.3. Supplied wrench.

3. At this point, the S.U.B. hardware is ready to be prepared for a cell

culture run.

4. For systems with load cell display screens, refer to Appendix B for
information about calibrating load cells.

CAUTION: Do not move the unit (especially when filled) while load cells
are unlocked, as this can damage the load cells.

5. To lock load cells that have been unlocked, hand-tighten the

lockout nut onto the post. Use the supplied tool to turn the nut an
extra 1/4 turn.

CAUTION: To avoid damaging the load cells, do not over-tighten the

nut. Assemble a standard stainless 38.1 mm (1.5 in.) tri-clamp around
the flanges. Complete this process for all load cells.

2.2.2 Leveling and connecting the system

All manual movements of mobile S.U.B. hardware should be over
smooth surfaces with the S.U.B. empty and disconnected from all
power and gas/feed sources. All load cells must be fully locked down
in order to move a S.U.B. Refer to the previous subsection of this guide
for illustrations.

1. Verify that the facility electrical supplies are sufficient to support the
power requirements of the S.U.B. and ancillary components, such
as controllers or pumps.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 36

Chapter 2 | Hardware assembly and setup

2. Locate the outer support container in the area for the cell culture
run.

3. When monitoring the batch volume, the unit may be placed on a

weight scale if load cells are not part of the system. Other methods
may be used to measure all incoming and outgoing liquids.

4. Level the platform by disabling the swivel casters on the bottom of

the outer support container. This is accomplished by threading the
leveling feet (at the center of each caster) to the floor.

5. Verify the location of the pH/DO controllers and ensure that the
cable and tubing lengths are sufficient.

WARNING: Risk of electrical shock.

6. Verify that the main power is off and the emergency stop is pulled
out. Note: The emergency stop disconnects all power to the
system. An alarm buzzer will sound when the emergency stop is
activated.

7. Verify that the main motor power switch is in the "off" position.

8. Connect all electrical plugs to facility power. Note: 120 VAC 250 L
S.U.B.s should be connected to a dedicated 20 A circuit. Refer
to hardware/electrical labels and schematics to ensure proper
electrical voltage is connected to the S.U.B. The main power switch
can now be turned on.

9. For 1,000 L units only: the water jacket ports are removed
for shipping. Attach the ports to the S.U.B. using the tri-clamps
provided (Figure 2.4).

Figure 2.4. Attaching water jacket port using tri-clamp.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 37

Chapter 2 | Hardware assembly and setup

10. Connect water inlet and outlet lines from the temperature control
unit quick-connects to the jacket (Figure 2.5). For 50, 100, 250,
500, and 2,000 L units, the inlet is typically on the left side if you
are facing the connectors. For the 1,000 L S.U.B. unit, the inlet is
the lower connection, and the outlet is the upper.

Inlet port

Outlet port

Figure 2.5. Inlet and outlet ports.

11. Insert the sparge line support (Figure 2.6) into the bottom of the
S.U.B. unit, directly below where the sparger will be placed. This
piece holds the sparge line vertically for maximum effectiveness.
The sparge line can be wound through the coil of the holder to
keep the sparger oriented properly.

Figure 2.6. Sparge line support.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 38

Chapter 2 | Hardware assembly and setup

2.2.3 Verifying drive shaft segments for 2,000 L systems

The 2,000 L S.U.B. is supplied with a special drive shaft that differs in
appearance and material when compared to the metallic shafts used
in smaller S.U.B. sizes. Due to the higher mechanical stress generated
in 2,000 L S.U.B.s, these systems require drive shafts made of carbon
fiber composites to reduce the weight of the long shaft.

Note: Always maintain a log history of the drive shaft and confirm
that it has sufficient life remaining. For warranty purposes, users
must show documentation of proper drive shaft use. A sample
log for documenting drive shaft use is provided in Appendix D of this
publication. If the age or history of a drive shaft is questionable, it
should be discarded.

2.2.4 Setting the air pressure rate for motor lift

adjustment (1,000 and 2,000 L systems only)

The air pressure rate must be set before using the handheld controller
for motor lifts on 1,000 and 2,000 L systems (see section 2.2.5). The
pressure regulator located on the back of the outer support container
(Figure 2.7) maintains the level of air pressure used to adjust the motor
lift.

Figure 2.7. Pressure regulator.

Hook up the pressure regulator to an air source and turn on the air.
Increase or decrease the air supply until the air pressure rate is at
90 psi.

WARNING: Air pressure for adjusting the motor lift on 1,000 and
2,000 L systems must stay at or below 90 psi. Air pressure above
90 psi may cause damage to the hardware.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 39

Chapter 2 | Hardware assembly and setup

2.2.5 Using the handheld controller for motor lifts

(1,000 and 2,000 L systems only)

The motor lift on 1,000 and 2,000 L systems is operated by a handheld

controller (Figure 2.8), which is located on a bracket attached to the
front of the S.U.B. Before use, refer to section 2.2.4 to ensure that the
air pressure rate is properly set.

Emergency stop

Position dial
Locked/unlocked
indicators Position indicators

Unlock/move
button

Figure 2.8. Handheld controller for

motor lifts.

WARNING: Rotating and moving parts, such as the motor lift for
1,000 and 2,000 L systems, can cause injury. Keep hands away from
moving parts during operation. Do not operate this equipment unless
the supplied guarding is in place and properly functioning. It is the
responsibility of the end user to assess this equipment and ensure that
equipment and safeguards are in good working condition. All operators
must be trained and aware of entanglement and pinch hazards and
associated protective devices, such as hazard signs and guarding.

Use the following instructions to change the motor position by

operating the handheld controller.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 40

Chapter 2 | Hardware assembly and setup

1. Before adjusting the motor lift, verify that the motor cap is closed.
If the cap is open, the motor will not run. The red status sensor on
the motor cap (Figure 2.9) communicates to the handheld controller
if the cap is open or closed.

Status sensor
on motor cap

Figure 2.9. Red status sensor on motor cap.

2. Hold down the unlock/move button on the handheld controller

to unlock the motor. The "unlocked" indicator on the handheld
controller will be lit orange (Figure 2.8). While holding down the
unlock/move button, turn the position dial to the left to lower the
motor position, or to the right to raise the motor position.

Note: The motor must be in either the "up" or "down" position.

If the motor is not in either position, the "intermediate position"
indicator will be lit red as a warning (Figure 2.8).

3. After the motor angle is in either the "up" or "down" position,

release the unlock/move button to lock the pin located on the side
of the motor. The "locked" indicator will be lit green.

Important notes:
• Prior to BPC loading, the motor should be in the "up" position.
• After the BPC has been loaded and filled with air, the motor should
be moved to the "down" position.
• To stop the handheld controller and movement of the motor lift in
the case of an emergency, use the red Emergency Stop (E-Stop)
button on the top of the handheld controller.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 41

 Operating information
3
Chapter contents
3.1 General system operating information
3.2 BPC and drive shaft loading instructions for 50, 100, and
250 L systems
3.3 BPC and drive shaft loading instructions for 500 and 1,000 L
systems
3.4 BPC and drive shaft loading, and condenser system setup
instructions for 2,000 L systems
3.5 Probe preparation and insertion
3.6 Cell culture operating instructions
3.7 Verification procedures

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 42

Chapter 3 | Operating information

3.1 General system operating information

3.1.1 BPC preparation

Each outer support container is designed for a specific BPC. Confirm

that the correct volume and type of BPC is being used for the
corresponding volume outer support container. 50–500 L 5:1 BPCs
are different than 2.1 BPCs, and are labeled for 5:1 mixing. Sections
3.2, 3.3, and 3.4 cover the installation and setup of BPCs. Follow these
instructions in the order in which they are presented.

3.1.2 BPC handling instructions

If you are using a sharp object when opening outer polybags, take care
to avoid damaging the BPC. Do not drag containers over corners or
sharp objects. Do not lift the container by the corners or top seams.
Carefully coil the tubing on top of the BPC to prevent puncturing the
container with cable ties or clamps. Use cushioning between the tubing
and the container in storage and transport.

3.1.3 BPC operating information

Working volume
Each S.U.B. is designed for a specific working volume range. The
minimum working volume and the rated working volume are listed in
the specification tables provided in Chapter 4 of this user's guide. The
total volume listed includes the headspace needed for proper aeration
and gas management.

Note: Actual working volumes should not exceed the indicated rated
working volumes by more than 10%. In addition, working volumes less
than 20% of the rated volume can result in damage to the BPC and/or
the S.U.B. hardware.

Operating pressure
The BPC does not operate as a closed system; it has both inlet and
exhaust filters that are utilized to maintain a sterile environment for cell
growth. However, conditions can be encountered when the gas inlet
flow rate may exceed the exhaust flow rate. This may be encountered
in the unlikely event of a pressure regulator failure on a gas feed, or
when excessive foam within the bioreactor creates a vent blockage.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 43

Chapter 3 | Operating information

WARNING: The BPC is not rated as a pressure vessel. Gas

pressure within the BPC headspace should not exceed 0.03 bar
(0.5 psi) at any time. Pressure above 0.03 bar (0.5 psi) may result in
BPC damage or personal injury.

• More demanding applications may warrant an optional exhaust vent

heater.
• If foaming is excessive in your cell culture process, it is best to
reduce the operating volume of the process to 80% of maximum
rated working volume of the S.U.B. system being used to provide
greater headspace volume.
• Single-use pressure transducers are available on custom S.U.B.
configurations. This technology combined with high-level control
systems (common with industrial applications) can regulate gas
pressure within the confines of the S.U.B.

Aeration
Gas to liquid mass transfer in cell culture bioreactors is controlled
by the solubility of the gas in the liquid, its distribution, and the
temperature and pressure. Direct air sparging provides for the oxygen
requirements of eukaryotic cell cultures. It allows optimal aeration of
the culture process and effective carbon dioxide stripping. However,
when compared to 2:1 mixing (50% working volume), 5:1 mixing causes
more carbon dioxide buildup in the extra headspace in the BPC when
operating at 20% working volume. This blanket of carbon dioxide may
prevent proper cell respiration.

A cross flow sparging strategy disrupts the dense carbon dioxide

blanket at the bottom of the BPC headspace, which lets users take
advantage of low-volume mixing without compromising cell cultures.
This strategy requires temporarily rerouting the overlay sparger to a
cross flow port near the surface of the liquid when operating at 20%
working volume. When the volume is increased above 20%, the sparge
line should be returned to the standard overlay port to reduce carbon
dioxide in the headspace, and a single drilled hole sparger is the main
source of aeration.

For more information about possible sparging strategies, review the

test data in the Thermo Scientific HyPerforma 5:1 S.U.B. Validation
Guide (DOC0023).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 44

Chapter 3 | Operating information

Aseptic connections
The most commonly recommended process for making connections
to the tubing lines is with an aseptic tubing fuser. Other connection
options are available as a custom BPC assembly. By following the
recommended tubing welder operating instructions, successful
connections can be made for filling, supplementing, sampling, or
dispensing from the BPC as needed.

Draining and harvest

The S.U.B. is equipped with a bottom drain line that allows for liquid
harvest by means of peristaltic pump. Connection of the bottom drain
line can be accomplished by use of a tubing welder or the fitting that is
provided. The bottom drain exits the BPC at the lowest vertical position
on the side of the S.U.B. This allows for easy access for the user and
minimizes the accumulation of cells in the area of the drain during the
cell culture run. Manipulation of the BPC as the last few liters of media
drain will minimize liquid hold-up within the S.U.B. The 2,000 L S.U.B. is
provided with a 25.4 mm (1 in.) bottom drain near the center line of the
tank bottom.

3.1.4 Hardware operating information

Heating performance
Heating times for 5:1 S.U.B. systems vary based on liquid volume and
temperature, ambient or heating liquid temperature, sparging rate, and
mixing rate. For heating times, see Table 3.1.

WARNING: Do not heat the system if the BPC is not at 20%

liquid volume or greater. Batch temperature should not exceed
40°C.

Table 3.1. Heating times for S.U.B. systems. Ambient temperature of 25°C—values
assume a TCU heater size of at least 9 W per batch liter.
Liquid batch volume Time
System Initial liquid Liquid target
(20% / 100%) (20% / 100%)
50 L 10 L/50 L 5°C 37°C 1 hr/1.1 hr
100 L 20 L/100 L 5°C 37°C 0.9 hr/1.6 hr
250 L 50 L/250 L 5°C 37°C 1.1 hr/3.4 hr
500 L 100 L/500 L 5°C 37°C 1.1 hr/2.2 hr
1,000 L 200 L/1,000 L 5°C 37°C 1.2 hr/4.1 hr
2,000 L 400 L/2,000 L 5°C 37°C 1.4 hr/4.0 hr
Note: Conditions may vary based on your system connections and environment.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 45

Chapter 3 | Operating information

Protective earth grounding (units with AC motors)

For units with AC motors, protective earth grounding for the S.U.B.
hardware system and the controller is provided through the ground
terminal of the power plug. Source power to the controller must
provide protective earth grounding to this terminal in order to minimize
the hazard of a possible shock in the occurrence of a fault condition.
Please refer to Appendix A for information about electrical receptacles.
A ground wire is provided underneath the S.U.B. and must be tied to
the controller before operation.

Agitation control interface for units with AC motors and E-Box

enclosures
The agitation control interface utilizes a digital display to indicate stirring
speed in units of revolutions per minute (rpm). Power is supplied to
the motor by a two-position power switch that is illuminated in green
when turned to the "on" position (right position). The agitation should
not be operated at volumes less than 20%. Stirring speed is adjusted
using the up and down arrows on the agitation keypad interface on the
E-Box, or using the settings on an integrated third-party controller. Due
to the auto-restart capabilities of the S.U.B., the green start button on
the keypad has been disabled; however, the red stop button on the
keypad is active.

If the red stop button has been used to stop the motor, the controller
can be reset and agitation restarted by using the main motor toggle
switch on the left side of the E-Box. For more information, see the
illustrations in the E-Box detail in section 4.3.

Circuit protection (units with AC motors)

Electrical components of the S.U.B. are equipped with circuit
protection. The variable frequency drive used to power the mixer
motor is protected by the use of a 10 A double pull resettable breaker
with a type C time delay (5-10 x LN). Other components, such as
the temperature controller and heating element, are protected with
resettable breakers.

In the case of an electrical fault condition, these safety devices

are designed to protect the user from electrical shock and prevent
electrical system components from being damaged. Fuses can be
replaced and/or the breakers reset once the fault condition is resolved.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 46

Chapter 3 | Operating information

Electrical breaker notes:

• The normal "on" setting for these breakers is in the up position.

• A tripped breaker will be in the mid position.
• The "off" setting is in the fully down position.
• To reset a tripped breaker, it must first be moved from the mid
position to the "off" setting (fully down position) before moving it to
the "on" setting (fully up position).

Scales and weighing systems

Monitoring liquid volume within the S.U.B. during operation can be
critical in cell culture applications that involve nutrient media feeds. This
can also be a useful method for increasing the scalability of the S.U.B.
by starting the process run at minimum operating volume. The ability
to track operating volume by use of load cells or weigh scales allows
the user the ability to control liquid volume and cell density as the
bioreactor is increased to rated working volume during the process run.

A load cell kit for weight/volume measurement is available for all S.U.B.
units, which can be installed at the factory or can be added later by
a certified service technician. The load cell kit comes with three load
cells, summing block, wiring, and display with a choice of several
interfaces.

Refer to Appendix B for load cell display calibration instructions.

Ensure that load cells are locked down before any movement of
the S.U.B. unit.

To lock the load cells before transporting any size S.U.B., follow the
steps below and refer to Figure 2.2 and Figure 2.3 in section 2.2.1.

1. Hand-tighten the load cell lockout nut onto the load cell lockout
post. You may need to use the small end of the supplied wrench to
loosen the load cell lockout nut from the bottom of the base.

2. After the nut is hand-tightened against the post, use the small end
of the supplied wrench to turn it an extra 1/4 turn.

CAUTION: To avoid damaging the load cell, do not overtighten

the nut.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 47

Chapter 3 | Operating information

3. Assemble a standard stainless 28.6 mm (1.5 in.) tri-clamp around

the flanges.

4. Repeat steps 1 through 3 for all load cells on the S.U.B.

3.1.5 External data logging and control

Digital display weighing scales can be sourced from manufacturers

such as Mettler Toledo. Bench top scales are commonly used to
measure the amount of bulk source media stored in a smaller-volume
BPC as it is transferred by peristaltic pump into the S.U.B.

Floor scales can be used to measure the fluid content within the S.U.B.
This is accomplished by rolling the S.U.B. onto the scale platform and
leveling the S.U.B. skid once in position.

The S.U.B. hardware systems are designed to allow advanced users to

control all aspects of the operation of the bioreactor. Contact technical
support for Thermo Scientific HyPerforma products general integration
guidance.

3.2 BPC and drive shaft loading instructions for

50, 100, and 250 L systems

3.2.1 Initial BPC loading steps for 50, 100, and 250 L
systems
Each outer support container is designed for a specific BPC. Verify
that the correct volume and type of BPC is being used for the
corresponding volume outer support container. The BPC should have a
tag on the bearing port labeled "5:1 Mixing." Use the following steps to
install and set up the BPC.

1. Remove the irradiated BPC from the protective double polybags

(Figure 3.1). Remove the cable ties from the drain line.

2. Load the BPC from the top into the outer support container,
avoiding any sharp edges that may damage the BPC (Figure 3.2).

3. Orient the BPC with the bearing port up and toward the motor drive
with the aseptic connector probe ports facing the bottom access
cutout.

4. Place the bearing port into the bearing port receiver, close the door
and close the clamp (Figure 3.3).
Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 48
Chapter 3 | Operating information

Figure 3.1. BPC removed from Figure 3.2. BPC loading. Figure 3.3. Bearing port insertion.
protective double polybags.

5. Use the back access window to route the side and bottom ports
through the opening in the outer support container (Figure 3.4).

6. Route the sparge lines, bottom drain, and sampling lines through
the appropriate openings (Figures 3.5 and 3.6). Note: Although
Figure 3.5 shows two drilled hole sparge lines, 50, 100, 250, 500,
and 1,000 L standard 5:1 BPCs only have one drilled hole sparge
line.

7. Route the sparge lines through the bottom plate and loop them
around the sparge line holder (Figure 3.5).

Figure 3.4. Bottom line access. Figure 3.5. Sparge line setup.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 49

Chapter 3 | Operating information

Figure 3.6. Drain line and port setup.

8. If a cable management system is available (see system shown

in Figure 3.7), attach the lines to the appropriate inlet ports
(Figure 3.8).

Figure 3.7. Cable management Figure 3.8. Incoming line connection

system in use. to inlet ports.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 50

Chapter 3 | Operating information

9. Connect the incoming gas feed lines to both the overlay filter and
the sparger filter. Ensure that the filters are located above the
maximum liquid level (Figure 3.9).

Figure 3.9. Inserting lines into the cable

management system channels.

10. Inflate the BPC with air through the overlay filter, but do not exceed
25 slpm or 0.034 bar (0.5 psi) internal BPC pressure. Inflation time
is approximately 10–20 minutes. Time will vary based on flow rate,
inlet pressure, and container volume. As the BPC inflates, ensure
that the ports, drain, and sparge lines are properly oriented in the
support container.

WARNING: The BPC is not rated as a pressure vessel. The BPC

should not be allowed to become tight during inflation or operation.
DO NOT EXCEED 0.034 bar (0.5 psi) within the BPC or the BPC could
fail. For reference, the BPC will appear to be tight at 0.007 bar (0.1 psi).
See Table 3.7 in section 3.6.8 for recommended air flow rates. The
operating pressures at the level of the S.U.B. are of primary importance
and these values must be adhered to.

11. As the container fills with air, check to make sure the sparge lines
are properly aligned. Note: Standard 5:1 S.U.B.s only use one
sparge line.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 51

Chapter 3 | Operating information

Sparge line note: While a sparge line check valve is provided

for each sparge line, it is not uncommon for some fluid to bypass
check valves during typical use. Elevating the filter will reduce the
chance that the filter is exposed to liquid.

12. Use the four bottom cutouts located at the base of the support
container as a reference to align the hanging tab on the BPC
(Figure 3.10).

CAUTION: Do not attach cutouts to any of the bottom hanging

tabs; only use them for reference in aligning the BPC within the
tank. Doing so may potentially stretch and/or tear the film.

Figure 3.10. Attaching hanging tab and hook.

13. Position the bottom side drain, pulling out and downward to
position the port toward the bottom edge of the S.U.B.

14. Align the row of probe ports within the access window (Figure 3.11).
Note: Verify all port clamps are closed and located as close as
possible to the body of the BPC.

Figure 3.11. Aseptic connector alignment.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 52

Chapter 3 | Operating information

15. Connect the media ground clip to the stainless steel insert in the
sample line on the BPC. This step grounds the media inside the
BPC and helps to eliminate electrostatic charge (Figure 3.12).

Ground clip
connection

Figure 3.12. Media ground clip connection.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 53

Chapter 3 | Operating information

3.2.2 Drive shaft insertion for 50, 100, and 250 L

systems

WARNING: Before you insert the drive shaft, the BPC must be
adequately inflated so that it is sitting upright in the outer support
container.

Figure 3.13 illustrates the components of the motor and mixing

assembly. The parts labeled on the figure will be referenced throughout
the drive shaft insertion process. Use the steps on the following pages
to insert the drive shaft.

Motor Drive shaft

cap head

Motor drive
keyway
Hollow
pass-
through Latch pin

Safety
cover Motor

Drive
shaft

Figure 3.13. Motor and mixing assembly.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 54

Chapter 3 | Operating information

1. Remove the latch pin from the safety cover over the mixing
assembly and open the cover. Unscrew the motor cap covering the
hollow pass-through of the motor (Figure 3.14).

Figure 3.14. Removing the motor cap.

WARNING: Ensure that the correct drive shaft is being used. Because
the 5:1 S.U.B. is capable of mixing at much lower volumes than
traditional 2.1 S.U.B.s, it is important to use the correct drive shaft to
avoid damaging the hardware. 50–500 L 5:1 drive shaft heads are
black and engraved with "5:1 Mixing." 2:1 mixing uses a system with
a different impeller angle, and is covered in a separate S.U.B. user's
guide.

2. Insert the drive shaft through the hollow pass-through of the motor
assembly in the following manner (Figures 3.15–3.18).
• Use two hands to load the drive shaft through the top of the
motor assembly; a slight back-and-forth twisting motion will aid
in insertion and avoid stretching the impeller tubing (Figures 3.15
and 3.16). Do not push the drive shaft straight in.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 55

Chapter 3 | Operating information

Figure 3.15. Loading drive shaft. Figure 3.16. Twisting drive shaft to aid
insertion.

• When approximately 50.8–76.2 mm (2–3 in.) of the shaft remains,

twist back and forth slightly to engage the impeller (Figure 3.17).
• When approximately 25.4–50.8 mm (1–2 in.) of the shaft remains,
twist back and forth slightly to engage the bearing assembly.
• When approximately 6.4 mm (0.25 in.) of the shaft remains, twist
to align the motor drive keyway with one of the four outer slots
on the drive shaft head (Figure 3.18).

Figure 3.17. Preparing to engage impeller. Figure 3.18. Drive shaft head aligned.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 56

Chapter 3 | Operating information

3. Directly couple the drive shaft to the motor drive (Figures 3.19–3.21).
• Place the motor cap on the hollow pass-through and
hand-tighten clockwise (Figure 3.19).

Figure 3.19. Replacing motor cap.

• Tighten the motor cap by placing a spanner wrench on the

hollow pass-through and tighten the motor cap using the
supplied torque wrench (Figure 3.20). Wrench note: The torque
wrench is a standard 10 mm (3/8 in.) square drive, and it is
calibrated at the factory at 16.9 Nm (150 in-lb.).

Figure 3.20. Tightening cap.

• Remove the wrenches from the system and place in the storage
holders.
• Close the safety access cover and insert the latch pin
(Figure 3.21).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 57

Chapter 3 | Operating information

Figure 3.21. Replacing and latching cover.

3.2.3 Final installation steps for 50, 100, and 250 L

systems

1. The air supply to the overlay can be turned off once the drive shaft
has been inserted.

2. Optional: Wrap and secure the vent filter heater on the exhaust filter.
Connect the heater to the controller and verify that it is plugged into
an appropriate 120 or 240 VAC outlet, then connect the power cord
to the controller. The controller is preset to 50°C.

3. Secure the exhaust vent filter on its holder (Figure 3.22).

Note: Some custom BPCs are supplied with dual exhaust vents.
The vent bracket can accommodate 10 in. and 4 in. filters in either
single or dual configuration.

Figure 3.22. Vent filter installation.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 58

Chapter 3 | Operating information

4. Attach the overlay sparge line and any other lines to the cable
management system, if available (Figure 3.23).

Figure 3.23. Optional cable

management system on a S.U.B. unit.

5. Position and close a bar clamp on the bottom drain line as close as
possible to the BPC port (Figure 3.24).

Figure 3.24. Bar clamp installation.

6. Remove the plastic insert located in the thermowell, if present.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 59

Chapter 3 | Operating information

7. Insert the RTD or selected temperature sensor into the thermowell

(Figures 3.25 and 3.26).
• Place a small amount of glycerol (0.5 mL) in the well to aid in
heat transfer. The glycerol also acts as a lubricant, which helps
with probe insertion.
• The sensor should be inserted until the base of the probe meets
the mouth of the thermowell. Rotate the probe either clockwise
or counterclockwise to aid insertion.
• Secure by twisting the luer lock collar, if provided. The
thermowell will stretch slightly when the RTD is seated.

Figure 3.25. Sensor insertion. Figure 3.26. Securing sensor.

8. Optional: Connect a pressure sensor to the CPC aseptic connector

at the top of the BPC. Then connect the appropriate pressure
transducer cable to the third-party controller.

9. Continue to section 3.5 for probe preparation and insertion

instructions.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 60

Chapter 3 | Operating information

3.3 BPC and drive shaft loading instructions for

500 and 1,000 L systems

3.3.1 Initial BPC loading steps for 500 and 1,000 L

systems
Note: The figures in this section feature a 500 L 5:1 S.U.B. Please
note that 1,000 L 5:1 S.U.B.s have a motor that can be operated in two
positions. The motor must be in the "up" position to load the BPC into
1,000 L S.U.B.s.

Checkpoints prior to BPC loading

9 The correct volume BPC is being used for the corresponding
volume outer support container. 500 L 5:1 BPCs should have a tag
on the bearing port labeled "5:1 Mixing."
9 The outer support container is stationary with the casters locked
into place. BPC loading may require operators to step inside the
bioreactor, and the unit must be stationary for the safety of both the
operator and equipment.
9 Two operators are available for ease in BPC loading.
9 A ladder or other means of elevation is available for drive shaft
insertion.
9 The motor is in the "up" position (1,000 L systems only). See section
2.2.5 for instructions on using the handheld controller to change the
position of the motor.

Use the following steps to install and set up the BPC.

1. Open the door on the bioreactor support container and reach

inside to open the clamp on the bearing port receiver located below
the motor (Figures 3.27 and 3.28).

Figure 3.27. Opening the bioreactor door. Figure 3.28. Close-up of bearing
receiver clamp.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 61

Chapter 3 | Operating information

2. Remove the irradiated BPC from the protective double polybags

(Figure 3.29). Do not remove the polybags from the line sets at this
stage as the BPC may become difficult to manage. Do not allow the
BPC or line sets to touch the floor.

3. Reach into or step inside the outer support container with the
front face (bearing port side) of the BPC oriented toward the motor
(Figure 3.30).

Figure 3.29. BPC in protective Figure 3.30. Bearing port orientation.

polybags.

4. Place the top line sets, still in polybags, over the top edge of the
tank (Figure 3.31). This will keep the container from being restricted
during the air inflation step.

Figure 3.31. Line sets on edge of tank.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 62

Chapter 3 | Operating information

5. Load the container bearing port into the receiver (Figure 3.32).
Close the door and clamp it shut (Figure 3.33).

Figure 3.32. Bearing port in receiver. Figure 3.33. Door clamped shut.

6. Remove the bubble wrap from the sparger filters. Guide the sparge
inlet lines and filters through the bottom cutouts in the outer
support container (Figure 3.34). The operator can reach just below
the S.U.B. to further extend the sparge lines from the cutouts
(Figure 3.35).

Figure 3.34. Sparge line insertion. Figure 3.35. Sparge line extension.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 63

Chapter 3 | Operating information

7. Pass the bagged drain line set and temperature/sampling port

set through the large cutout in the front of the outer support
container (Figure 3.36). Extend the drain line set through the cutout
(Figure 3.37).

Figure 3.36. Drain/sampling line set Figure 3.37. Drain/sampling line

insertion. extension.

8. Connect the pressure transducer to the monitor. After the display

has stabilized, tare the monitor. The monitor should be allowed to
warm up for 30 minutes before taring. Verify that the monitor reads
zero.

9. The BPC must be partially inflated until it is sitting upright. This

allows proper insertion of the drive shaft and aids in the proper
alignment of the BPC in the outer support container.
• Attach the air supply to the overlay gas inlet line. Note: Air
pressure to the overlay gas line on the S.U.B. BPC should not
exceed 0.2 bar (3 psi) or 25 slpm.
• Begin air inflation through the overlay gas line. Filling the
container with air takes approximately 15–20 minutes before
drive shaft insertion can begin. Times will vary based upon flow
rate and inlet pressure.
• Steps 10–13 can be completed while the BPC is filling with air.

WARNING: The BPC is not rated as a pressure vessel. DO NOT

EXCEED 0.034 bar (0.5 psi) within the BPC or the system could fail,
causing personal injury or damage to equipment. DO NOT leave the
BPC unattended while inflating. See Table 3.7 in section 3.6.8 for
recommended air flow rates. The operating pressures at the level of the
S.U.B. are of primary importance and these values must be adhered to.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 64

Chapter 3 | Operating information

10. Attach the incoming gas supply to the sparger gas inlet line. Note:
Air pressure to the sparger on the BPC should not exceed 0.55 bar
(8 psi). Also, while a sparge line check valve is provided for each
sparge line, some fluid may bypass check valves during typical
use. Elevating the filter to ensure that it is not at the low point of the
sparge line will reduce the chance that the filter is exposed to liquid.

11. Tare the load cell display before proceeding.

12. Attach all of the hanging tabs on the BPC to the hooks on the
bottom of the outer support container to help position the ports
(Figures 3.38 and 3.39).

CAUTION: For the 500 L systems, only attach the front two
hanging tabs to the pins to assist in aligning the probe belt, drain
port, and spargers.

Figure 3.38. Hanging tab and hook. Figure 3.39. Attaching tab.

13. Verify that the sparger filter and spargers remain in the correct
position. It is recommended that users secure only the front two
hanging tabs on the front BPC panel first. This way the door will not
be an obstruction when connecting the last set of hooks.

14. Remove the protective packaging from the exhaust vent filters
(Figure 3.40).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 65

Chapter 3 | Operating information

Figure 3.40. Removing protective packaging.

15. If you are using a 1,000 L system, move the motor into the "down"
position using the handheld controller after the BPC has filled with
air. See section 2.2.5 of this manual for detailed instructions for
using the handheld controller.

3.3.2 Drive shaft insertion for 500 and 1,000 L systems

The drive shaft is constructed in multiple segments, which must be

assembled and inserted in pieces. Operators should be elevated (i.e.
ladder) to effectively assemble and insert the drive shaft.

CAUTION: Review ceiling height requirements in Chapter 4 of this

user's guide before trying to insert the drive shaft.

WARNING: Ensure that the correct drive shaft is being used for 500 L
systems. Because the 5:1 S.U.B. is capable of mixing at much lower
volumes than traditional 2.1 S.U.B.s, it is important to use the correct
drive shaft to avoid damaging the hardware. 500 L 5:1 drive shaft
heads are black and engraved with "5:1 Mixing." 2:1 mixing in 500 L
S.U.B.s uses a system with a different impeller angle, and is covered
in a separate S.U.B. user's guide. 1,000 L 5:1 systems use drive shafts
with white heads that are NOT labeled for 5:1 mixing.

Figure 3.41 illustrates the components of the motor and mixing

assembly. The parts labeled on the figure will be referenced throughout
the drive shaft insertion process. Use the steps in this section to
assemble and insert the drive shaft.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 66

Chapter 3 | Operating information

Motor Drive shaft

cap head
Motor drive
keyway
Hollow
pass-
through Latch pin

Safety
cover Motor

Drive
shaft

Figure 3.41. Motor and mixing assembly.

1. Prepare the hollow pass-through by first removing the latch pin on

the safety cover (Figure 3.42), opening the safety cover (Figure 3.43)
and removing the motor cap of the mixing assembly (Figure 3.44).

Figure 3.42. Opening safety cover. Figure 3.43. Cap removal.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 67

Chapter 3 | Operating information

Figure 3.44. Latch pin removal.

2. Verify that the two, three, or four segments of the drive shaft, all
with matching serial numbers, are located in the drive shaft holders
on the side of the outer support container. For the three-piece
drive shaft loading described here, the segments will be referred
to as upper (the segment with the drive shaft head), middle (the
segment with the internal/external threads on each end), and lower
(the segment with the square end). For 1,000 L systems, lubricate
the threaded ends with a light coat of food-grade anti-seize with
each use. Always verify that the drive shaft segments have
matching serial numbers prior to assembly and use.

3. First, insert the lower segment through the hollow pass-through

of the mixer drive (Figure 3.45). Slide the latch pin from the motor
assembly into the shaft to prevent it from falling into the tube
(Figure 3.46). Assemble the middle and lower segments of the
drive shaft by joining them with a twisting motion, fastening the two
segments together (Figure 3.47).

Note: Segmented shafts are left-threaded (reverse-threaded) to

avoid loosening during operation.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 68

Chapter 3 | Operating information

Figure 3.45. Inserting lower section. Figure 3.46. Latch pin in shaft.

Figure 3.47. Segment assembly.

4. Place one wrench on the flat area in the middle drive shaft segment
and another wrench on the lower segment, then tighten the
connection using a counterclockwise rotation (Figure 3.48). After
the segments are secure, return the wrenches to the tool holder.
CAUTION: Do not over-tighten; a snug fit is sufficient. Remove the
latch pin.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 69

Chapter 3 | Operating information

Figure 3.48. Tightening of shaft

connections.

5. Repeat the actions in steps 3 and 4 to attach the upper segment of

the drive to the partially-assembled drive shaft.

6. Using two hands, carefully guide the completed drive shaft into the
BPC using a slight back-and-forth twisting motion. Note: It may be
necessary for another operator to assist with drive shaft insertion.
As one operator inserts the drive shaft, another operator should
carefully manipulate the impeller as the end of the drive shaft
begins to couple with the impeller. Do not push the drive shaft
straight in.
• When 50.8–76.2 mm (2–3 in.) of the shaft remains, twist slightly
to engage the impeller (Figure 3.49).

Figure 3.49. Drive shaft insertion.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 70

Chapter 3 | Operating information

• When 25.4–50.8 mm (1–2 in.) of the shaft remains, twist slightly

to engage the bearing assembly.
• When 6.35–12.7 mm (0.25–0.50 in.) of the shaft remains, twist
to align the motor drive keyway with one of the four outer slots
on the drive shaft head (Figure 3.50).

Figure 3.50. Drive shaft head aligned.

7. Directly couple the drive shaft to the motor by placing the motor
cap back on the hollow pass-through and tighten.

8. Tighten the motor cap by placing the spanner wrench

counterclockwise on the hollow pass-through and tighten using the
supplied torque wrench (Figure 3.51). Wrench note: The torque
wrench is a standard 10 mm (3/8 in.) square drive and is calibrated
at the factory at 16.9 Nm (150 in-lb.).

Figure 3.51. Tightening motor cap with wrenches.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 71

Chapter 3 | Operating information

9. Verify that the wrenches are removed from the system and returned
to the storage holders.

10. Close the safety access cover and insert the latch pin.

3.3.3 Final installation steps for 500 and 1,000 L

systems
1. Secure the exhaust vent filters to the top-mounted holders
(Figure 3.52), or if you are using elevated dual exhaust filters, use
the adapter piece and extended filter bracket (Figure 3.53). Note:
500 L BPCs and some custom BPCs are supplied with dual
exhaust vents. The vent bracket can accommodate 10 in. and 4 in.
filters in either single or dual configuration.

Figure 3.52. Vent filter. Figure 3.53. Extended dual filter bracket.

2. Fully extend the drain line set through the front cutout and attach
the probe shelf.

3. Remove the polybag from the drain line set and position the line
clamp as close as possible to the BPC port and close. Use a cable
tie around the clamp to ensure the clamp does not open.

4. Align the aseptic connector ports through the front access window
(Figure 3.54).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 72

Chapter 3 | Operating information

Figure 3.54. Aseptic connector port alignment.

5. Remove the plastic insert located in the thermowell, if present.

6. Insert the RTD or another selected temperature sensor into the

thermowell (Figure 3.55).

• Place a small amount of glycerol (0.5 mL) in the thermowell to aid

in heat transfer. The glycerol also serves as a lubricant and aids
in insertion.
• The sensor should be inserted until the base of the RTD meets
the mouth of the thermowell.
• Secure by twisting the luer lock collar, if provided; the thermowell
will stretch slightly when the RTD is seated (Figure 3.56).

Note: Verify that all port clamps are closed and located as close as
possible to the body of the BPC.

Figure 3.55. Temperature sensor insertion. Figure 3.56. Securing temperature

sensor.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 73

Chapter 3 | Operating information

7. Optional: Connect a pressure sensor to the CPC aseptic connector

at the top of the BPC. Then connect the appropriate pressure
transducer cable to the third-party controller.

8. Refer to section 3.5.3 for probe insertion instructions.

9. Close the bottom access door. The proper latch tension can be
obtained by a combination of feel and visual inspection. When
closing the latch, the handle should begin to provide resistance
to closing when the leading edge of the safety pin pass-through
of the latch handle aligns with the outside edge of the latch base
(Figure 3.57). Note: When the latch is under-tensioned, the safety
pin pass-through of the latch handle will be covered within the
latch base and the handle will close very easily. If the latch is over-
tensioned, the handle will be excessively difficult to close.

Safety pin pass-

through of latch
set

Latch retainer

Figure 3.57. Latching the access door.

10. The access doors must be closed and fully latched prior to filling
the system with liquid.

11. Continue to section 3.5 for probe preparation and insertion

instructions.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 74

Chapter 3 | Operating information

3.4 BPC and drive shaft loading, and

condenser system setup instructions for
2,000 L systems
3.4.1 Initial BPC loading steps for 2,000 L systems

Checkpoints prior to BPC loading

9 The correct volume BPC is being used for the corresponding
volume outer support container.
9 The motor is in the "up" position. For information on using the
handheld controller for motor lifts, see section 2.2.5.
9 The BPC lift control is plugged into a 120–240 VAC receptacle, and
the Main Disconnect and Motor Enable switches on the BPC lift
control are switched to "off."
9 Three operators are available for ease in BPC loading.
9 A ladder or other means of elevation is available for drive shaft
insertion (see the specifications in Chapter 4 for system ceiling
height requirements).
9 The equipment has been evaluated against your confined space
safety standards and procedures.

Use the following steps to install and set up the BPC.

1. Switch on the main power to the control panel. Ensure that the
drive motor is not running. Open both the front and rear doors on
the outer support container.

2. Verify that the motor area is clear and the Vertical Lift switch on
the BPC lift control (located on the side of the S.U.B.) is in the "up"
position. Then flip the Motor Enable switch on the BPC lift control to
the "on" position. The BPC lift should perform a homing operation,
which fully raises the wires to the hard stop of the support arms
before lowering them to the "up" position.
• CAUTION: Keep hands clear during the homing operation.
• The homing operation is complete when the lift stops moving.

3. Before lowering the BPC lift, ensure that the S.U.B. area is clear.
Do not lower the BPC lift if there are any obstructions. Flip
the Vertical Lift switch down to lower the BPC lift hooks.

4. Two operators should carefully remove the irradiated BPC from

the protective double polybags (Figure 3.58). Do not remove the
polybags from the line sets at this stage as the BPC may become
difficult to manage. Do not allow the BPC or line sets to touch or
drag on the floor.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 75

Chapter 3 | Operating information

5. Load the BPC through the rear access door (Figure 3.59), orienting
the bottom of the container into the door first with the bearing
port facing upward. Keep the container folded as supplied in the
packaging to allow the BPC to unfold naturally when it is raised by
the BPC lift.

Figure 3.58. Removing the BPC from polybags. Figure 3.59. Loading the BPC.

6. Using the rear or front door for access, connect the retainer hooks
on the BPC lift to the top of the BPC via two of the hanging tabs
(located across from each other) (Figure 3.60).

CAUTION: While operating the 2,000 L bag hoist with heavier

customer BPCs, watch for excess stress on the BPC during
lifting. Reposition the BPC as necessary to avoid tearing the
BPC hanging tabs.

Figure 3.60. Connecting tabs to BPC lift.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 76

Chapter 3 | Operating information

7. One operator should remain elevated to observe from above while

another operator, at ground level, operates the BPC lift control. Flip
the Vertical Lift switch up on the BPC lift control to raise the BPC
lift hooks. CAUTION: Do not reach into the tank during this
operation. The BPC lift should remain in the "up" position during
the rest of the BPC loading process.

8. After the BPC has been lifted, hook the tabs on the bottom of the
BPC to the pins on the S.U.B.

9. Use an elevated platform to open the clamp on the bearing port

receiver located underneath the motor. Remove the black protective
cap from the bearing port (Figure 3.61), load the BPC bearing port
into the receiver (Figure 3.62), close the bearing assembly door, and
latch it (Figure 3.63).

Figure 3.61. Removing the bearing port cap. Figure 3.62. Loading the bearing port.

Figure 3.63. Closing and latching door.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 77

Chapter 3 | Operating information

10. Place the top line sets (still in polybags) over the side of the outer
support container (Figure 3.64). This will help support the weight of
the BPC and also keep the BPC from being restricted during the air
inflation step.

Figure 3.64. Placing linesets over tank edge.

11. If your BPC includes exhaust vent filters, ensure that they are lifted
through the middle of the motor lift instead of to either side. This
will ensure the exhaust vent filters are properly oriented. If you are
using the exhaust condenser system, follow the setup instructions
in section 3.4.2 of this guide. If you are using elevated exhaust vent
filters, use the corresponding extended dual vent filter bracket and
filter heaters.

To load the optional exhaust vent filters, follow the steps below.

• Clip each filter one at a time into the elevated vent filter holder
system (Figure 3.65). Carefully center the filter housing, allowing
the clip to secure it near the hose barb connections.

Figure 3.65. Clipping filter to

holder.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 78

Chapter 3 | Operating information

• Ensure that the routing of the exhaust tubing will not likely
become kinked.
• Place the vent heaters around each filter (Figure 3.66), verifying
that the snap retainers are secured. Position the power leads to
avoid interfering with the vent holder brackets.

Figure 3.66. Installing heaters.

• Raise and rotate the vent holder bracket as needed (Figure 3.67).
Make a final inspection to ensure that no kinks or low spots will
occur in the tubing between the BPC and the filter, even if the
BPC becomes pressurized.

Figure 3.67. Raising the vent holder

bracket.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 79

Chapter 3 | Operating information

• Connect the power to the vent heaters and verify operation of

the controllers.
• Inspect the controllers' setpoints (recommended 60°C). After
two to five minutes of operation, verify that the vent heaters are
warm and are near the desired temperature setpoints. Verify that
no alarm indicators are active.

12. Open the tubing set polybag and connect the pressure transducer
to the monitor. Once the display has stabilized, tare the monitor.
The monitor should be allowed to warm up for 30 minutes and
the sensor connected for 10 minutes before taring. Verify that the
monitor reads zero.

13. Clamp the drilled hole sparge, exhaust, and cross flow lines so that
the air supplied by the overlay gas inlet line flows directly into the
BPC (Figure 3.68).

Figure 3.68. Clamping the exhaust lines prior to

filling the BPC with air.

14. The BPC must be partially inflated to aid in the proper alignment of
the BPC in the outer support container, and proper insertion of the
drive shaft.

• Attach the air supply to the overlay gas inlet line at the top of
the BPC.
• Begin filling the BPC with air. Operators should allow the
container to fill to greater than half volume. This typically takes
less than 20 minutes.
• Steps 15–18 can be completed while the BPC is filling with air.

Note: Air pressure to the overlay gas line on the BPC should not
exceed 0.344 bar (5 psi) or 100 slpm.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 80

Chapter 3 | Operating information

WARNING: The BPC is not rated as a pressure vessel. DO NOT

EXCEED 0.034 bar (0.5 psi) within the BPC or the system could fail,
causing personal injury or damage to equipment. DO NOT leave the
BPC unattended while inflating. See Table 3.7 in section 3.6.8 for
recommended air flow rates. The operating pressures at the level of the
S.U.B. are of primary importance and these values must be adhered to.

15. Feed the probe belt, sample line, and the subsurface addition lines
through the front access door (Figure 3.69).

Figure 3.69. Feeding lines through the front access door.

16. Remove the sparge lines from the polybags and bubble wrap from
the sparge filters. Use the rear door to gain inside access to the
floor of the hardware. Place a clamp on the bottom drain line at this
time (Figure 3.70).

Figure 3.70. Clamping bottom drain line.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 81

Chapter 3 | Operating information

17. The center insert on the tank floor provides the port locations for
the bottom drain (Figure 3.71) and the gas lines for the drilled hole
sparger. Guide the sparger inlet line and filter through the bottom
cutout in the tank (Figure 3.72). To remove the bottom cutout, lift
and rotate it in a counterclockwise direction.

Figure 3.71. Drain line support. Figure 3.72. Drilled hole sparger line.

Sparge line notes:

• The plastic center tank insert also contains cutouts for porous
frit sparge lines. Porous frit spargers are not available on
standard 5:1 BPCs, but they are optional on custom BPCs.
• Air pressure to the spargers on the BPC should not exceed
8 psi.
• While a sparge line check valve is provided for each sparge
line, it is not uncommon for some fluid to bypass check valves
during typical use. We recommend elevating the sparge line
filter as is feasible to help reduce this tendency.

18. Attach all of the hanging tabs to help position the ports. Using
the positioning tab pins provided, secure the BPC by attaching
the tabs on the bottom of the BPC onto the position tab pins
(Figures 3.73 and 3.74).

19. Verify that the sparger filter and spargers remain in position while
attaching the tabs. It is recommended that users secure the tabs
on the front BPC panel first. This way, the larger rear door will allow
access when connecting the last set of tab pins.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 82

Chapter 3 | Operating information

Figure 3.73. Pulling the container tab toward the Figure 3.74. Securing the container tab on the pin.
pin.

20. After the BPC has filled to greater than half volume, unclamp the
drilled hole, exhaust, and cross flow lines.

3.4.2 Condenser system setup for 2,000 L systems

Condenser system functional overview

The condenser system is intended to be used as an accessory for
large S.U.B.s as an alternative to vent filter heaters. Condenser systems
are recommended for use with 2,000 L S.U.B.s. The condenser’s
purpose is to prevent liquids and solids from condensing and collecting
inside of the vent filters of the S.U.B. The condenser system cools
the exhaust gases leaving the S.U.B. chamber, condensing the
moisture out of the saturated gases coming from the S.U.B. The liquid
condensate that is stripped from the exhaust gases is then pumped
back into the bioprocessing container chamber, creating a sterile
loop and significantly reducing liquid loss due to evaporation. The
condenser plate on condenser systems with a cart assembly is chilled
by a closed bath recirculating chiller, which has sufficient capacity to
cool two condenser plates simultaneously. The condenser plate on
side-mounted condenser systems is chilled by a house recirculating
chilling loop. Figures 3.75 and 3.76 show both the cart assembly and
side-mounted (2,000 L systems only) condenser system options.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 83

 Chapter 3 | Operating information

Complete
condenser system
Condenser disposables Condenser hardware

Filter bracket
Exhaust line
Filter straps assembly
from S.U.B.
Condenser Condenser post
Exhaust plate assembly assembly
vent
filters Dual
Post receivers
headed
Condenser peristaltic
return line pump
Condenser back to S.U.B.
bag gas Closed bath
outlet port Condenser recirculating
Cart
bag gas chiller
assembly
Dual chamber inlet port
condenser bag
Gripping
tabs
Condenser bag Alignment
liquid drain ports holes

Figure 3.75. Overview of condenser system cart assembly option for 2,000 L S.U.B.s.

Figure 3.76. Side-mounted condenser

system option for 2,000 L S.U.B.s.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 84

Chapter 3 | Operating information

When to use the condenser system

2,000 L S.U.B. BPC with single-use condenser system
Large 254 mm (10 in.) hydrophobic PVDF filters with a nominal 0.2
micron pore size were specified in order to increase the available
surface area for off-gassing. In conjunction, the standard 2,000 L
S.U.B. is designed for use with a single-use condenser system. This
allows the S.U.B. to utilize a powerful phase-change type system which
provides improved exhaust vent protection and reliability due to the
ability to strip condensate and atomized materials that may be present
from the off-gas stream of the S.U.B. This system has been shown to
significantly reduce the “fouling” load on the vent filters that inherently
increases operating back pressure as the cell culture run batch
progresses. See the HyPerforma 5:1 S.U.B. Validation Guide for details.

2,000 L S.U.B. BPC with vent and heaters only

Some end users may prefer to omit the condenser system on the
2,000 L S.U.B. with the expectation that this will allow for a more
uniform installation (similar to smaller S.U.B. systems used in the
upstream seed train), or will perhaps reduce system complexity and
cost. The use of exhaust vent heaters and 254 mm (10 in.) filters will
provide impressive flow capacity over short periods (less than five
days). However, the high sparge rates required during the scale-up
of the S.U.B. to the 2,000 L working volume may eventually create
conditions of increased operating back pressure, usually due in part to
blocking of the filter media. Depending upon the application, the user
has the option of using both filters in parallel or initiating the run with a
single filter, temporarily clamping off the line to the other filter (it being
reserved as redundant back-up).

Table 3.2 may help end users specify the BPC configuration and
operating parameters for custom 2,000 L S.U.B. applications when
not utilizing the exhaust condenser. Because the operating parameters
of different cell cultures vary widely, a safety factor should be used to
temper the data. Accordingly, the data we used to generate a control
base line are for reference only (filter fouling will vary and must be
considered to ensure reliable performance). It is assumed no foam is
present in the exhaust stream.

Table 3.2. Condenser system overview.

S.U.B. system Maximum combined flow Resulting safety
Run duration
2 each 254 mm (10 in.) vents rate recommended factor
2,000 L S.U.B. 7 days 40 slpm 2x
2,000 L S.U.B. 10 days 32 slpm 2.5x
2,000 L S.U.B. 14 days 27 slpm 3x
2,000 L S.U.B. 21 days Single-use condenser strongly recommended

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 85

Chapter 3 | Operating information

The above recommendations were generated using the test

conditions shown in Graph 3.1. In this case, a 2,000 L S.U.B. was
filled with 2,000 liters of DI water with a batch temperature of 40°C
using an MKS vent filter heater at 60°C. Safety factor estimates are
based on a maximum continuous internal S.U.B. BPC pressure not
to exceed 0.006 bar (0.1 psi), which corresponds to 40 slpm with
a single 254 mm (10 in.) vent. Note: These results do not take into
consideration a “fouling” safety factor.

Graph 3.1. 2,000 L S.U.B. pressure trending.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 86

Chapter 3 | Operating information

Also consider the size and type of tubing used to connect the exhaust
vents to the S.U.B. BPC (when not using an exhaust condenser).
Braid reinforced tubing provides the best protection against kinking
or accidental pinching of the exhaust line. The 254 mm (10 in.) vents
are supplied with 19.1 mm (0.75 in.) hose barbs and this tubing
diameter will allow condensate to return to the S.U.B. at total off-
gas flow rates up to 30 slpm (assuming the tubing is near a vertical
orientation). Testing has shown that large diameter tubing will allow for
lower exhaust gas velocities and if the vapor velocity is below 0.6 m/s
(1.96 ft./s), gravity will allow the condensate formed in the tubing to
return to the batch process (Graph 3.2).

Note: Restrictive tubing connectors can create flow bottle necks and
12.7 mm (0.5 in.) inner diameter (ID) tubing is typically deemed too
small for the 2,000 L S.U.B.

Graph 3.2. Tubing diameter recommendation.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 87

Chapter 3 | Operating information

Various vent filter configurations are available on the S.U.B. depending

upon the process scale and intended application. Graph 3.3 provides
a reference for determining the relative capacity of different filters
depending upon the amount of gas flow anticipated and the length of
the run. In all cases, using a vent filter heater will reduce the chance of
condensate blocking the filter, but over time, suspended solids carried
in the exhaust stream will impede the flow of exhaust gas (resulting in
increased back pressure). In addition, it is good practice to monitor
the amount of foam present in the head space. In all cases, a vent
filter heater has very little tolerance for handling the presence of foam
in the exhaust stream. A small feed of antifoam (e.g., FoamAway™
Irradiated AOF Antifoaming Agent, catalog number A1036901) added
directly to the liquid surface of the culture head space typically provides
excellent foam control. 1,000 and 2,000 L systems can benefit from
the use of a condenser system. It has been shown to increase system
reliability at high flow rates (beyond 50 slpm) and should warrant strong
consideration when performing batch runs beyond 10 days. Results
will vary; however, it is strongly recommended that end users select a
vent filter configuration providing reserve capacity where possible. For
example, dual vent configurations can be used independently with the
second filter serving as a redundant backup (providing a quick reserve
in case issues arise in process).

Graph 3.3. Off-gassing management guidelines.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 88

Chapter 3 | Operating information

Condenser system setup

1. Remove the reservoir cap of the chiller and add the appropriate
type and volume of fluid as per the chiller user's guide.

2. Verify that the peristaltic pump and chiller power cords are
connected to a power source.

3. If you are using a condenser system with a cart assembly, plug in

the system.

4. Turn on the power to the chiller. This will allow the chiller to prime.

5. If you are using a temperature control unit (TCU), ensure that the
TCU coolant is filled to the maximum level.

CAUTION: Low TCU coolant levels can increase the temperature

of the plates, and cause excessive pressure and/or residue buildup
in the condenser bag. Please note that the chiller plates may run
warmer than the TCU setpoint.

6. Purge the chill plate by loosening the bleed plug on top of the
plate. This is accessed using a hex wrench passing through the top
tensioning plate of the chill plate assembly. Loosen the plug only
enough to allow trapped air to escape, then re-tighten.

7. The settings for the chiller and peristaltic pump are preset at the
factory. These settings allow for the system to resume setpoint if
the power is temporarily disrupted. Verify that the chiller and pump
setpoints are at the recommended levels (5°C and 12 rpm).

8. If you are using a condenser system with a cart assembly, verify

that the peristaltic pump is in place on the cart beneath the chill
plate. Side-mounted condenser systems have an attached tray for
the peristaltic pump.

Condenser system loading

Two operators are required to safely set up the exhaust system. Setup
time is typically 2–3 minutes.

Note: The figures in this section show a condenser system with a cart
assembly. Side-mounted condenser systems have the same chiller
plate and use the same loading instructions.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 89

Chapter 3 | Operating information

1. One operator, located at an elevated position, should remove

the condenser BPC carefully from the polybag packaging. Lower
the assembly (directed in a vents-first orientation) to the second
operator located at ground level, standing to the rear of the S.U.B.
For systems with a cart assembly, the second operator should
stand between the condenser cart and the S.U.B.

2. The operator at the upper position should move to ground level,

open both doors on the chiller plate, and load the condenser BPC
from the front, keeping the BPC in a saddle bag shape. Allow the
vents to hang freely (Figure 3.77).

Figure 3.77. Opening the chiller plate doors.

3. Route the gas inlet lines around and behind the vent holders, and
inspect both lines to ensure they are connected to the S.U.B. and
are not twisted or kinked, adjusting them as needed (Figure 3.78).

Figure 3.78. Routing gas inlet lines.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 90

Chapter 3 | Operating information

4. The second operator should hold the vent filters and place them
into the vent filter holders above the chill plate (Figure 3.79).

Figure 3.79. Placing vent filters.

5. The first operator should use the Velcro™ straps to secure the filters
in position (Figure 3.80).

Figure 3.80. Securing vent filters.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 91

Chapter 3 | Operating information

6. Use the grasping tabs to position the container using the two lower
button pins on each side of the chiller plate (Figure 3.81).

Figure 3.81. Positioning container.

7. Close the clear side doors while carefully manipulating first the
gas inlet line and then the gas outlet line (Figure 3.82) to clear the
doors as each is closed and latched (Figure 3.83).

Figure 3.82. Moving gas lines. Figure 3.83. Latching door.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 92

Chapter 3 | Operating information

8. As the doors are closed and latched, the second operator should
route the gas inlet lines into the clips behind the vents (Figure 3.84).

Figure 3.84. Clipping gas lines into

place.

9. The first operator should load the peristaltic tubing into the pump
(located on the cart for condenser systems with a cart assembly,
or on the tray for side-mounted condenser systems), verifying
that there is sufficient slack at each end of the pump tubing. Then
align the tubing in the pump channel and close the pump ramp
(Figure 3.85).

Figure 3.85. Loading pump tubing.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 93

Chapter 3 | Operating information

10. Start the pump by pushing the red button (Figure 3.86). Verify that
both the pump and chiller are enabled and running at the proper
settings. We recommend setting the pump at 12–30 rpm and the
chiller at 5°C. The specified pumping system is qualified to run
continuously (wet or dry) beyond 21 days.

Figure 3.86. Starting the pump.

11. After setup, verify the following:

9 The elbow fittings on the inlet and outlet of the condenser

saddle bag are straight and level.
9 The gas inlet line and the condensate line are not twisted,
pinched, or obstructed.
9 There are no low spots in the gas inlet line. Adjust the lines to
avoid condensation pooling.
9 The pump union is loose on both ends of the pump and running
smoothly in the peristaltic rollers.

Contact technical support for specific condenser system performance

questions.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 94

Chapter 3 | Operating information

3.4.3 Drive shaft insertion for 2,000 L systems

The new drive shafts for 2,000 L S.U.B.s are slightly longer and have
black drive shaft heads. Thermo Fisher Scientific has redesigned
the impeller in all standard 2,000 L S.U.B. BPCs to improve impeller
engagement with this new drive shaft design. 2,000 L 5:1 BPCs are
shipped with a hanging tag on the bearing port showing the required
drive shaft length (Figure 3.87).

Figure 3.87. Close-up view of

2,000 L 5:1 BPC hanging tag.

The drive shaft is constructed by assembling two quick-connect

segments. Operators should be elevated (i.e. ladder) to assemble and
insert the drive shaft.

CAUTION: Review ceiling height requirements for the drive shaft in the
specifications in Chapter 4.

Figure 3.88 illustrates the components of the motor and mixing

assembly. The parts labeled on the figure will be referenced throughout
the drive shaft insertion process.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 95

Chapter 3 | Operating information

Motor
cap Drive shaft
head

Motor drive
keyway
Hollow
pass-
through Latch pin

Safety
cover Motor

Drive
shaft

Figure 3.88. Motor and mixing assembly.

Use the following steps to insert the drive shaft.

1. The BPC must be filled with air to greater than approximately 50%
volume to allow for unrestricted loading of the angled drive shaft.
Note: After inflation, the impeller tubing should be hanging straight
down inside the BPC, with the impeller near the bottom.

2. Verify that the proper drive shaft segments and tools are available.

3. Prepare the hollow pass-through by first removing the latch

pin on the safety cover (Figure 3.89), opening the safety cover
(Figure 3.90), and removing the threaded cap by turning it
counterclockwise. Use the wrench and spanner provided if
required.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 96

Chapter 3 | Operating information

Figure 3.89. Removing latch pin. Figure 3.90. Opening safety cover.

4. Verify that the two segments of the drive shaft have matching serial
numbers, and are located in the drive shaft holders on the side of
the outer support container.

Important notes: Always verify that the drive shaft segments have
matching serial numbers prior to assembly and use. No lubrication
is required with the quick-connect assembly design.

5. First, insert the lower segment through the hollow pass-through of

the mixer drive (Figure 3.91). Once inserted, slide the latch pin from
the mixing assembly into the shaft to prevent it from falling into the
impeller sleeve (tube) (Figure 3.92).

Figure 3.91. Insertion of the lower Figure 3.92. Using the latch pin.
section of the drive shaft.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 97

Chapter 3 | Operating information

6. To connect the upper and lower sections together, depress the

button on the female side (Figure 3.93) and slide the sleeve back.
This will expose a red ring underneath the sleeve (Figure 3.94). This
is a visual indicator that the sleeve is not in a locked position.

Figure 3.93. Button used for Figure 3.94. Sliding sleeve exposing
connection. red "not locked" indicator.

7. Place the female side of the quick-connect over the male end
(Figure 3.95). The connection is fully seated when the red indicator
ring (Figure 3.96) on the male end is no longer exposed.

Figure 3.95. Quick connection. Figure 3.96. Red indicator ring exposed.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 98

Chapter 3 | Operating information

8. Slide the sleeve toward the connection, allowing the push button
to lock into position. This will engage the locking mechanism and
also cover the red indicator ring (Figure 3.97). Note: When fully
connected, no red coloring should be visible.

Figure 3.97. Sliding the sleeve into place.

9. Once the sections are secure, remove the latch pin and return the
wrenches to the tool holder.

10. Using two hands, carefully guide the completed drive shaft
into the BPC using a slight back-and-forth twisting motion or a
counterclockwise rotation (Figure 3.98). Do not push the drive
shaft straight in.

• When 50.8–76.2 mm (2–3 in.) of shaft remains, twist slightly to

engage the impeller.
• When 25.4–50.8 mm (1–2 in.) of shaft remains, twist slightly to
engage the bearing assembly (Figure 3.99).
• When 6.4 mm (0.25 in.) of shaft remains, twist to align the motor
drive keyway with one of the four outer slots on the drive shaft
head.

Figure 3.98. Insertion of the drive shaft. Figure 3.99. Engaging the bearing port.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 99

Chapter 3 | Operating information

11. Ensure that the head is fully seated before directly coupling the
drive shaft to the motor. Any spring-back indicates that the drive
shaft is not properly seated in the impeller. Figure 3.100 illustrates a
drive shaft that is completely inserted into the impeller.

Drive shaft end

Figure 3.100. Drive shaft fully inserted into the

impeller.

Note: The cap should be easy to install when the drive shaft head
is fully engaged in the hollow pass-through. Otherwise, repeat
steps 1 through 13 before installing the cap.

12. Place the threaded cap back on the hollow pass-through. Secure
the cap by placing a spanner wrench on the hollow pass-through
and tightening using the supplied torque wrench (Figure 3.101).

Figure 3.101. Tightening cap.

Note: The torque wrench is a standard 10 mm (3/8 in.) square drive,

and it is calibrated at the factory at 16.9 Nm (150 in-lb.).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 100

Chapter 3 | Operating information

13. Verify that the wrenches have been removed from the system and
returned to the storage holders.

14. Close the safety access cover and insert the latch pin.

3.4.4 Final installation steps for 2,000 L systems

1. Verify the proper position of the exhaust filters. The exhaust flow
path must be unobstructed. Connect the gas supply lines. Verify
the intended flow paths for overlay, cross flow, and drilled hole
spargers.

2. Verify that the overlay and direct sparger lines are correctly
positioned and free of kinks. Verify that the rear access door is
closed with proper latch tension.

3. Remove the polybag from the drain line set and verify that the
redundant line clamps are in position. Use a cable tie around the
clamp to ensure the clamp cannot be accidentally opened.

4. Align the aseptic ports through the front access window

(Figure 3.102).

Figure 3.102. Aseptic port alignment.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 101

Chapter 3 | Operating information

5. Secure the access doors with the latches. Proper tension is

obtained by adjusting the threaded latch pin. Tension of the latch is
adjusted by varying the position of the pin on the threaded shank.
The proper latch tension can be obtained by a combination of feel
and visual inspection. When closing the latch, the handle should
begin to provide resistance to closing when the leading edge of the
safety pin pass-through of the latch handle aligns with the outside
edge of the latch base (Figure 3.103).

Note: When the latch is under-tensioned, the safety pin pass-

through of the latch handle will be covered within the latch base
and the handle will close very easily. If the latch is over-tensioned,
the handle will be excessively difficult to close.

Safety pin
pass-through Safety pin pass-
of latch-handle through of latch set

Latch retainer Outside edge of latch

base

Figure 3.103. Latch access door.

6. For maximum security, insert pins (not included) into the respective
latches.

7. Turn off the air supply to the overlay line.

8. Position clamps as close as possible to the BPC and close them on

all tube ports (Figure 3.104).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 102

Chapter 3 | Operating information

Figure 3.104. Clamp installation.

9. Place clamps on subsurface lines as close to the port as possible.

This will eliminate media from filling these lines prior to use.

10. Remove the plastic insert located in the thermowell, if present.

11. Insert the RTD or selected temperature sensor into the thermowell
(Figure 3.105).

Figure 3.105. Insertion of the RTD into the

thermowell.

• Place a small amount (0.5 mL) of glycerol (Sigma-Aldrich™

G6279) in the thermowell to aid in heat transfer.
• The sensor should be inserted until the base of the RTD meets
the mouth of the thermowell.
• If provided, secure by twisting the luer lock collar. The
thermowell will stretch slightly when the RTD is seated.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 103

Chapter 3 | Operating information

12. Connect the batch-to-tank grounding cable to the stainless steel

connector of the sample line (Figure 3.106).

Figure 3.106. Grounding cable connected.

Note: Verify that all of the port clamps are closed and located as close
as possible to the body of the BPC.

13. IMPORTANT: During media fill, verify the position of all critical
ports (drain, spargers, line sets, and probes) before the container
is filled with more than 50 liters of liquid. This will still allow time for
adjustments, if required, after the fill is initiated.

14. Typically, two fill lines (12.7 mm [0.5 in.] x 19.1 mm [0.75 in.]) and
peristaltic pumps are recommended to fill the 2,000 L S.U.B. in a
timely manner.

3.5 Probe preparation and insertion

3.5.1 Preparation and sterilization

1. Select the appropriate probe (see section 1.3.1). Verify the presence
of a Teflon™ support ring and O-ring on the probe and visually
inspect the probe for damage.

2. Perform any required probe maintenance and calibrate the pH

probe (see section 3.5.4 for probe calibration information).

3. Insert the probe into the probe assembly through the threaded
adapter.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 104

Chapter 3 | Operating information

4. Verify that the probe tip is not touching (more than 6.35 mm
[0.25 in.] gap) the membrane of the aseptic connector before
threading into the probe adapter.

5. Hand-tighten the adapter and verify that the probe tip is not
touching the membrane.

6. Place the probe assembly with probe into the autoclave tray for
probe kits (Figure 3.107).

Handle

Probe
assembly

Autoclave tray
for probe kits

Figure 3.107. Probe assembly and autoclave tray.

7. Autoclave the probe assembly using a validated sterilization cycle

(approximately 30 minutes at 122°C). A 30-minute sterilization
cycle is generally sufficient. Options of wet or dry cycle parameters
can be used. Slow exhaust cycles are preferred, as this minimizes
stress on the probes during the temperature and pressure changes
of autoclaving.

8. Allow sufficient time for the probe assembly to cool completely

before connecting to the BPC for probe insertion.

9. When stored properly, autoclaved probe assemblies can be stored

dry for short periods of time (less than 24 hours) without loss of
sensor longevity, performance, or sterility.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 105

Chapter 3 | Operating information

3.5.2 Making CPC AseptiQuik connections

CPC AseptiQuik G genderless connector components

Figures 3.108 and 3.109 below illustrate the components of CPC™
AseptiQuik™ G genderless connectors. Connectors with white
protective cover pull tabs may be autoclaved. Generally, connectors
with blue pull tabs are gamma irradiated, not autoclaved. Visit the
Colder Products Company website at http://cpcworldwide.com for
more information.

For instructions on making an aseptic connection, see the following

section.

Paper
membrane

Hinge

Protective cover
pull tabs

Figure 3.108. CPC Aspetiquik G Figure 3.109. CPC Aspetiquik G

connector (closed). connector (open).

CPC AseptiQuik connection instructions

The following steps outline the process of making a sterile aseptic
connection using CPC AseptiQuik G genderless connectors.

1. Tear open and remove the plastic covering on the connector

located on the BPC (Figures 3.110 and 3.111).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 106

Chapter 3 | Operating information

Figure 3.110. Pulling the tear strip. Figure 3.111. Removing plastic covering from
connector.

2. Unsnap and flip open the protective cover pull tabs on both
connectors (Figures 3.112 and 3.113).

Figure 3.112. Opening protective cover pull tab Figure 3.113. Opening protective cover pull tab on
on port. lineset.

3. Align the two connectors and push them together.

4. Squeeze each side of the connectors until you hear a click

(Figure 3.114).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 107

Chapter 3 | Operating information

Figure 3.114. Squeezing connectors together.

5. Grab the joined pull tabs and pull upward to remove the paper
membranes from the connectors (Figure 3.115). The pull tabs will
also be removed.

Figure 3.115. Pulling tabs to remove paper membranes.

3.5.3 Probe insertion

Before beginning probe insertion, please become familiar with the

aseptic connector procedure outlined in section 3.5.2.

1. Attach probe clips onto the outer support container above the
probe assembly (Figure 3.116). Plastic probe clips slide on with firm
pressure.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 108

Chapter 3 | Operating information

Figure 3.116. Attaching plastic probe clip.

2. Install the pre-sterilized sensor and probe kit using the aseptic
connection methods described in section 3.5.2. The aseptic
connection is completed prior to the bellows being collapsed.

3. Insert the probe by collapsing the bellows (Figures 3.117 and 3.118).
Note: If the BPC is already filled with liquid, the best practice is to
squeeze the bellows to expel air prior to collapsing it. Then insert
the probe fully, as described.

Figure 3.117. Probe insertion. Figure 3.118. Collapsed bellows.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 109

Chapter 3 | Operating information

4. Position the probe clip in the desired horizontal location. Lift the
probe and set it into the probe clip (Figure 3.119).

Bellows hook

Figure 3.119. Positioning probe clip.

5. Rest the probe in the bellows hook. Release the probe assembly
and verify that the probe remains at the proper insertion depth and
angle when the bellows expand to rest freely in the probe clip.

3.5.4 Probe calibration

Probe calibration is controller-specific; however, the following general

rules apply:
• If you are using a liquid batch-to-tank grounding cable with the
stainless steel connector of the sample line, the sample line should
be purged of air prior to probe calibration.
• pH probes must be calibrated prior to steam sterilization; the
calibration of the probe can be standardized by comparison of
an off-line sample once the pH probe has been connected to the
S.U.B.
• Dissolved oxygen probes are generally calibrated after steam
sterilization. They can be calibrated once the probe is connected
to the S.U.B. and is given time to polarize (six to eight hours of
continuous connection to the power supply provided by a controller
or polarization module).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 110

Chapter 3 | Operating information

3.6 Cell culture operating instructions

3.6.1 Operating conditions for cell culture applications

Optimal operating parameters for cell culture vary greatly between cell
lines and media formulations. Table 3.7 in section 3.6.8 is provided
as a reference for establishing safe upper operating control limits with
the standard BPC design. Exceeding these operating limits may result
in premature exhaust filter failure, excessive foaming, and excessive
pressure build-up in the gas delivery line sets or the BPC.

In many cell culture operations, the limits listed in Table 3.7 are
excessive and should be further reduced when possible. When
reducing gas flow rate limits, the following trade-offs should be
expected.
• Reducing drilled hole sparger maximum operating limits will reduce
system foaming but increase reliance on O2. A suggested gas
operating control strategy is to run the drilled hole sparger on air
initially, and after total flow rate limits are reached, substitute oxygen
as shown in Graph 3.5.
• Reducing overlay maximum operating limits will reduce the exhaust
load (increasing filter lifespan) but will sacrifice sparger performance
if reduced far enough to allow CO2 buildup in the headspace.

If cell culture density is not increasing at expected rates, this may

be due to CO2 buildup in the headspace. Increasing air flow in the
headspace may resolve this problem.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 111

Chapter 3 | Operating information

3.6.2 Checkpoints prior to media fill

Verify the following before proceeding to liquid fill.

9 The BPC has been loaded into the hardware by following the
instructions provided in sections 3.2, 3.3, or 3.4.
9 All aseptic connector port heavy-duty clamps are closed and
located as close as possible to the BPC.
9 The exhaust filter is upright and secured using the holder.
9 The clamp on the drain tube is closed and located as close as
possible to the BPC.
9 The temperature RTD is completely seated in the thermowell and
secured.
9 The air-filled BPC is properly oriented in the outer support container
and the BPC bottom tabs are secured.
9 The gas line sets are connected to the drilled hole sparger and
either the cross flow sparger (for 5:1 operation) or overlay sparger
(for 2:1 operation).
9 All gas filters are placed above the maximum liquid level.
9 The load cell display has been tared.
9 The motor is in the "down" position (1,000 and 2,000 L systems
only).
9 All sensors are inserted and connected to their respective
transmitters. Sensors must be properly oriented to ensure that they
are below the liquid level after media fill.

3.6.3 Media fill

1. Select the desired line set from the BPC for fluid introduction.

2. Make an aseptic connection (tubing welder, quick-connect, or

tri-clamp) and begin liquid fill.

3. After no more than 20 liters have been added, verify the position of
the BPC in the outer support container, particularly the sparger and
the drain line. Adjust positioning if necessary for proper fit.

4. Pull the top corners of the BPC upward to reduce wrinkles during
filling. Note: If the BPC wrinkles are not eliminated during liquid fill,
excessive film tension below the bearing port will result.

5. Fill to the desired liquid volume (20–100% of the rated volume is

recommended).

6. Ensure that all sensors are below the liquid level after the BPC has
been filled.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 112

Chapter 3 | Operating information

3.6.4 Agitation for units with electrical control panels

1. After the media has reached the desired liquid volume, use the
motor controller power switch to start the agitation using the
electrical control panel (E-Box) (Figure 3.120).

Power reset Pressure display

switch
Temperature display

Motor
controller Motor speed control
power keypad
switch
Emergency
stop button

Main switch

Figure 3.120. Front view of E-Box for 50–2,000 L S.U.B.s.

2. Using the arrow keys on the motor speed control keypad, adjust the
setpoint speed to the desired level. The adjustment of the stirring
speed rpm is done using Hz. The display reverts back to displaying
rpm after 2–3 seconds of inactivity. Adjust desired agitation rate
within the recommended range as described in Table 3.3.

3. Allow the speed to stabilize, then make fine adjustments if

necessary.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 113

Chapter 3 | Operating information

Table 3.3. Recommended agitation rates. The values given are based on a standard scale-up criteria of
power input to working volume (P/V) using an estimated power impeller number of 2.1, not the parameters of
the motor. For rated maximum and minimum operating speeds for the motor, see the hardware specifications
in Chapter 4 of this user's guide. For information on calculating agitation rates using power input to volume
ratios, see section 3.6.5—Agitation Rate Calculations.

3
Nominal agitation 3 Impeller
Power 10 Watts per meters 40 Watts per meters
20 Watts per meters
3
diameter
ratio (rates in rpm) (rates in rpm)
(rates in rpm) (cm)
working

working

working

working

working

working

working

working

working
volume

volume

volume

volume

volume

volume

volume

volume

volume
100%

100%

100%
50%

50%

50%
20%

20%

20%
50 L 85 115 145 107 145 183 N/A 183 230* 11.1
100 L 68 92 116 85 116 146 N/A 146 184 14.6
250 L 54 74 93 69 93 117 N/A 117 148 20.0
500 L 47 64 80 59 80 101 N/A 101 127 25.1
1,000 L 39 53 67 50 68 86 N/A 86 109 32.1
2,000 L 35 47 59 44 60 75 N/A ** ** 39.8
* This value is outside the recommended operating motor range of 30–200 rpm (VFD settings). See Table 4.1 in Chapter 4.

** Consultation with Thermo Scientific engineers is required.

WARNING: Agitation must be stopped when the liquid level falls

below 20% of the rated working volume, otherwise damage to
the hardware or BPC may result. Do not exceed a 20 W/m3 power
ratio when operating between 20% and 50% working volume. See the
Warnings, Safety, and Warranty Information section in the front of this
publication, and section 3.6.5 for more information about using safety
interlocks and agitation speed governors on the controller.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 114

Chapter 3 | Operating information

3.6.5 Agitation rate calculations

Using power input to volume ratios for agitation speed

The power input to volume ratio (P/V) allows for scale-up or
scale-down of bioreactor platforms by equating mixing power between
differently-sized systems. P/V is the most broadly accepted method
for determining a practical scale-up approach for agitation speed in
stirred tank reactors used for animal cell culture. This relationship is
dependent on the density of the liquid in the system (ρ), the inherent
power number of the impeller (Np), the diameter of the impeller (Di), the
volume of liquid in the vessel (V), the speed of the drive shaft in rpm (n),
and the power input to the liquid (P).

This relationship can be rearranged to solve for mixing speed as a

function of the other variables.

For known values of P/V, impeller power number, density, and impeller
diameter, this equation can be simplified by consolidating all other
values into a single coefficient (A).

Values for appropriate agitator drive shaft speed were calculated for
different sizes of S.U.B.s using 20–100% of the rated working volumes.
These values assume an impeller power number of 2.1, viscosity similar
to water, and a constant density of 993 kg/m3.

Values of 20 watts/meter3 (W/m3) provide the nominal rating (the

suggested default parameter for CHO cultivation), and values of
40 W/m 3 provide the maximum recommended parameter that has been
qualified for use in the system. Graph 3.4 is provided as a reference,
showing the 20 W/m 3 P/V curve. While some discrete values are
shown, in practice it is acceptable to operate at lower speeds to suit
special needs.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 115

Chapter 3 | Operating information

3
Graph 3.4. Nominal agitation values using P/V of 20 W/m .

Fed batch cultures present unique challenges to the operation of

the 2,000 L S.U.B because the fluid height can change dramatically
through the duration of a fed batch cell culture run. This drastic change
in operating fluid height requires a very long agitator shaft supporting
the impeller, requiring it to sustain significant forces of rotating torque
and deflection of stress. The 2,000 L S.U.B. was originally qualified to
operate reliably when using discrete working volumes of either 1,000
or 2,000 L with a maximum recommended operating speed of 75 rpm.
3
This represented a worst case P/V of 40 W/m (1,000 L) and best case
3
of 20 W/m (2,000 L) as the extra volume and column height at 100%
working volume offer significant protection against drive shaft instability.

An in-depth study of the 2,000 L S.U.B. has shown that harmonics

can begin to propagate fatigue into the drive shaft when the system is
operated between 1,100 and 1,500 L when the agitator P/V is above
3
20 W/m . In order to maximize shaft life and reliability, it is important
to scale P/V with this constraint in mind. See Graph 3.5 for values of
potential cavitation and harmonics. It is recommended to operate your
S.U.B. at agitation values below the green curve.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 116

Chapter 3 | Operating information

Graph 3.5. Regions of potential agitator harmonics and cavitation for various liquid
3
working volumes of the 2,000 L S.U.B. The green line represents a P/V of 20 W/m . It is
recommended to operate2,000
2,000 LL S.U.B.
S.U.B.sAgitation
below thisGuidelines
curve.
100

90
Region of potential Region of potential
agitator cavitation agitator harmonics

80

70
Agitation (RPM)
60

50

40

30

20

10

0
0 200 400 600 800 1000 1200 1400 1600 1800 2000 2200
Volume (L)

20W/m^3
20 W/m3

P/V agitation values for all S.U.B. sizes

Equation coefficient values for every size of S.U.B. for P/V values of 20
and 40 W/m 3 are provided in Table 3.4 below.
3
Agitation speeds for P/V values of 20 and 40 W/m are provided in
Tables 3.5 and 3.6.

Table 3.4. Equation coefficient values (A) for different S.U.B.s.

n = A x V1/3 (for values of n in rpm)
50 L 100 L 250 L 500 L 1,000 L 2,000 L
S.U.B. S.U.B. S.U.B. S.U.B. S.U.B. S.U.B.
P/V
20 W/m3 49.650 31.491 18.634 12.764 8.471 5.950*
40 W/m3 62.554 39.676 23.477 16.081 10.673 7.031**
3
* Coefficient value is used to determine operating speeds for drive shafts implemented for a 180-day working duration at 20 W/m .
3
** Consult Thermo Scientific engineers for the drive shaft working duration at > 20 W/m .

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 117

Chapter 3 | Operating information

Table 3.5. Agitation rates (in rpm) for S.U.B.s at a P/V of 20 W/m3 .
50 L 100 L 250 L 500 L 1,000 L 2,000 L
S.U.B. S.U.B. S.U.B. S.U.B. S.U.B. S.U.B.*
Fill %
100 183 146 117 101 86 75
95 180 144 115 100 83 74
90 177 141 113 98 82 72
85 173 138 111 96 80 71
80 170 136 109 94 79 70
75 166 133 107 92 77 68
70 162 130 104 90 75 67
65 158 127 102 88 73 65
60 154 123 99 85 71 63
55 150 120 96 83 69 61
50 145 116 93 80 68 60
45 140 112 90 78 65 57
40 135 108 86 75 62 55
35 129 103 83 71 60 53
30 122 98 79 68 57 50
25 115 92 74 64 53 47
20** 107 85 69 59 50 44
* Using these rpm values requires replacement of the drive shaft after 180 days of use.

** These rpm values assume the system is operating at no less than 20% volume. In order to ensure proper volume measurements, end
users are responsible for ensuring proper load cell calibration prior to system use.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 118

Chapter 3 | Operating information

Table 3.6. Agitation rates (in rpm) for S.U.B.s at a P/V of 40 W/m
3
.
50 L 100 L 250 L 500 L 1,000 L 2,000 L
S.U.B. S.U.B. S.U.B. S.U.B. S.U.B. S.U.B.
Fill %
100 230 184 148 127 109 *
95 227 181 145 125 105 *
90 222 178 143 123 103 *
85 218 174 140 121 101 *
80 214 171 137 118 99 *
75 209 167 134 116 97 *
70 205 164 131 113 95 *
65 200 160 128 111 92 *
60 194 155 125 108 90 *
55 189 151 121 105 87 *
50 183 146 117 101 86 *
45 177 141 113 98 82 *
40 170 136 109 94 79 *
35 162 130 104 90 75 *
30 154 123 99 85 71 *
25 145 116 93 80 67 *
20** 135 107 86 75 64 *
3
* Consultation with Thermo Scientific engineers is required when operating 2,000 L S.U.B.s at a P/V of > 20 W/m .

** These rpm values assume the system is operating at no less than 20% volume. In order to ensure proper volume measurements, end
users are responsible for ensuring proper load cell calibration prior to system use.

Note: The impeller power number assumption of Np=2.1 is only an

approximation, based on an academic exercise. Recommended
best practice for determination and comparison of Np between
bioreactors should be carried out by the end user and should be based
on conditions matching the specific application. End users should
also anticipate and accept some variability that may be inherent in
the analysis technique or modeling method chosen. If a different or
alternative Np value is determined by the end user, the safety interlocks
must be based solely on the maximum prescribed rpm for a working
volume as found in the reference tables in this publication. Never rely
strictly on an estimated magnitude of P/V when setting the
agitator speed interlock levels.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 119

Chapter 3 | Operating information

3.6.6 Drive shaft rotation

Verify that the drive shaft is rotating counterclockwise when viewed

from the top looking down. The S.U.B. is designed to mix in this
direction only.

3.6.7 Temperature control

Temperature setpoints are controlled by the TCU or controller. Refer to

the TCU or controller manufacturer’s guidelines for setup and operating
instructions.

1. Connect to an external TCU using the large couplings located

on the vessel jacket. Ensure the inlet/outlet ports are connected
properly; improper installation may result in poor heating/cooling
performance.

2. Open the valves after connecting the TCU (Figure 3.121).

Figure 3.121. Opening valves.

Note: The water jacket should be purged of air any time the vessel
jacket lines are reconnected. To purge the water jacket, open the bleed
valve located near the bottom of the S.U.B. A container may be needed
to catch any glycol that is released. Close the valve as soon as glycol
begins flowing.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 120

Chapter 3 | Operating information

3.6.8 Sparging strategy

Sparging strategy and gas supply setup

Standard 5:1 BPCs are supplied with drilled hole, overlay, and cross
flow spargers. Figure 3.122 illustrates the location of each sparger on a
standard 500 L BPC. The cross flow sparger is used to reduce carbon
dioxide buildup in the headspace near the liquid surface when the
liquid volume is at 20%.

Front face

Cross flow
sparger
Overlay sparger

Back face

Figure 3.122. 500 L BPC spargers. Drilled hole

sparger

Graph 3.6 depicts a dissolved oxygen (DO) management strategy.

Refer to Table 3.7 in the following section for gas flow rate
recommended maximum values. In developing a gassing strategy for
a S.U.B. with a drilled hole sparger configuration, it is optional to have
a cross over from nitrogen to air when progressing from negative to
positive DO control. Graph 3.6 depicts a minor crossover.

When progressing into higher positive DO control, the drilled hole

sparger system is designed to maintain optimal oxygen delivery to
carbon dioxide stripping rates. When drilled hole sparger flow rate limits
are reached, supplant the sparger air flow with a steadily increasing
ratio of oxygen to allow a higher degree of control.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 121

Chapter 3 | Operating information

Graph 3.6. Drilled hole sparger DO control strategy for 5:1 S.U.B.
systems.

Gas flow rates

Standard 5:1 BPCs are supplied with drilled hole, overlay, and cross
flow spargers. If foaming or exhaust filter load/lifespan is of primary
concern, priority should be placed on tuning the system to operate
primarily by adding oxygen through the drilled hole sparger. If carbon
dioxide stripping is of primary concern, priority should be placed on
running the drilled hole sparger at flow rates sufficient to reduce or
eliminate the need for base addition.

Table 3.7 contains a listing of operating parameters for all sizes of

S.U.B.s. This data may be used in specifying maximum gas flow
rates for mass flow controllers or rotameters when using drilled hole
spargers. In optimal conditions, without condensation or fouling, the
exhaust filters have a flow capacity of at least 20 and 90 slpm (standard
liters per minute) at 0.006 bar (0.1 psi) for the small and large standard
equipped filter types. The total flow rate of gas into the system must be
less than the sum flow rate capacity of active exhaust filters.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 122

Chapter 3 | Operating information

Table 3.7. Range of operating parameters.

50 L 100 L 250 L 500 L 1,000 L 2,000 L
Temperature
2.0–40.0 ± 0.1
(°C)
OperatL) 10–50 20–100 50–250 100–500 200–1000 400–2000
Recommended

Overlay

Overlay

Overlay

Overlay

Overlay

Overlay
Cross

Cross

Cross

Cross

Cross

Cross
flow

flow

flow

flow

flow

flow
DHS

DHS

DHS

DHS

DHS

DHS
max. gas flow
rates (slpm)
Air 5 5 5 10 9 9 25 13 13 50 25 25 100 40 40 200 60 60
O2 5 - - 10 - - 25 - - 50 - - 100 - - 200 - -
CO2 1 - - 2 - - 2 - - 2 - - 5 - - 5 - -
N2 1 - - 2 - - 5 - - 5 - - 10 - - 10 - -
Total 5 5 5 10 9 9 25 13 13 50 25 25 100 40 40 200 60 60
Exhaust load 20 20 90 90 180 360

The values listed take into account the number and type of exhaust
filters that are standard on each size of S.U.B. (one small filter installed
in the 50 and 100 L, one large filter installed in the 250 and 500 L,
and two large filters installed in the 1,000 and 2,000 L vessels). These
values are not absolute requirements. They are also not intended to
be process gas flow settings. The process gas flow settings should be
adjusted as discussed below, with starting conditions not exceeding
25% of the listed maximum values to prevent unnecessary reduction of
exhaust filter life span and foam generation.

3.6.9 pH probe calibration

If employing a liquid batch-to-tank grounding cable with the stainless

steel connector of the sample line, the sample line should be purged of
air prior to probe calibration.

In general, the pH probe calibration (post-autoclave) can be verified by

pulling a sample and analyzing the pH on another calibrated pH meter.

3.6.10 DO probe calibration

After polarizing for six to eight hours, the DO probe can be calibrated
in the S.U.B. using standard protocols. Turn on full air sparging. Do not
exceed the maximum gassing rates listed in Table 3.7 in section 3.6.8.
Sparge until the DO stabilizes. Use the stabilized value as the 100% DO
setpoint for the controller. Set the zero percent DO setpoint by sparging
with nitrogen, or by temporarily unplugging the DO probe.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 123

Chapter 3 | Operating information

3.6.11 Checkpoints prior to inoculation

Before inoculation, verify that:

9 After the S.U.B. is filled with media to 20% or more volume, BPC
tabs have been disconnected from the S.U.B. hardware.
9 The pH probe is calibrated, autoclaved, and connected via an
aseptic connector port. Perform 1-point offset on your controller as
necessary.
9 The DO probe is autoclaved, connected via an aseptic connector
port, polarized (six to eight hours), and calibrated.
9 The temperature RTD is completely seated in the thermowell and
secured.
9 Ensure that all sensors are below the liquid level.
9 Operating parameters (temperature, agitation, pH, and DO) are at
the desired setpoints.
9 A method for making aseptic tubing connections is available.
9 Air is connected to the cross flow sparger when operating at 20%
volume.

3.6.12 Cell inoculation

Once the S.U.B. is operating at the targeted steady equilibrated state

and has achieved the proper temperature, the S.U.B. is ready for
inoculation. Connect the inoculum addition line set to the seed culture
vessel (equipped with the proper connectors/tubing) and transfer the
inoculum into the S.U.B.

Typically this is done with the tubing connection process (aseptic

luer lock connection or tube welding) and peristaltic pump. Pump the
desired volume of seed cells into the S.U.B.

Note: For shear sensitive cultures, cells can be introduced by

manipulating the addition port to direct the inoculum down the interior
wall of the BPC and into the bulk fluid, reducing the shear on the cells.
Custom line sets can be supplied with dip tubes, which shorten the
distance between the point of inoculum introduction and the bulk fluid
level.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 124

Chapter 3 | Operating information

3.6.13 Volume scale up

1. Using a sterile process, connect media to the BPC with the media
fill port.

2. Begin pumping media into the BPC at the desired flow rate. Ensure
that the vessel temperature does not drop below culture limits.

3. Remove the air line from the cross flow sparger, replace onto the
overlay gas port, and clamp the cross flow sparger port as close to
the BPC as possible.

4. Increase the volume to the desired level.

3.6.14 In-process checkpoints

Verify the following once or twice daily during the culture run.
9 Rising bubbles are visible through the access window.
9 Process parameters, such as temperature and agitation, are at
setpoint.
9 The BPC is not operating under pressure.
9 The cap is tight on the drive shaft.
9 The temperature sensor/RTD is completely seated and secured.
9 No condensate accumulates in exhaust filter housing. Accumulated
condensate indicates that the use of a filter heater is required.

Noise note: Noise may be emitted from the mixing assembly during
operation. This noise may vary in intensity and frequency, but generally
has no significant effect on performance or overall durability of the BPC
during the intended life of the product.

3.6.15 BPC sampling

During operation of the S.U.B., samples may need to be taken for
monitoring of various parameters established by the user. The following
sections describe two techniques for sampling: aseptic sampling with a
sterile syringe and sampling with a sterile manifold.

Aseptic sampling
Using a standard luer lock on a 60 mL syringe or manifold:

1. Remove the dust cover from the SmartSite needle-free valve, which
is connected to the end of the sample port (Figure 3.123).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 125

Chapter 3 | Operating information

Figure 3.123. Removal of dust cover from SmartSite.

2. Clean the SmartSite with a sanitary wipe.

3. Connect the sanitary luer lock type syringe (Figure 3.124).

Figure 3.124. Connecting the syringe.

4. To purge the sample line, apply a small amount of vacuum pressure

by pulling out the syringe plunger slightly.

5. Open the pinch clamp and pull a sample (approximately 20 mL),

using care not to allow any back flow.

6. Close the pinch clamp and remove the syringe. This will be a purge
sample.

7. Clean the SmartSite with a sanitary wipe.

8. Connect the sanitary luer lock type syringe.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 126

Chapter 3 | Operating information

9. Pull the sample by applying a small amount of vacuum pressure

using the syringe.

10. Open the pinch clamp and pull the desired sample volume
(approximately 10–20 mL), taking care not to allow any back flow.

11. Close the pinch clamp and remove the syringe. This will be a
representative sample.

12. Clean the SmartSite with a sanitary wipe and replace the dust cap.

Sampling with a sterile manifold

Use the steps below to attach a sample manifold (if purchased):

1. Remove the manifold from its protective polybag package.

2. Close all of the clamps on the manifold lines.

3. Use a sterile tubing welder to connect the manifold to the sample

line (Figure 3.125).

Figure 3.125. Tubing welder.

4. Inspect the welds and open flow path by pinching the welds.

5. Open two clamps at the inlet and the clamp at the purge container
(100 mL container).

6. Purge the sample line by filling the purge container (30–60 mL is

recommended).

7. Close the clamp nearest to the purge container.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 127

Chapter 3 | Operating information

8. Open the clamp to the sample container (50 mL container)

(Figure 3.126).

Figure 3.126. 50mL manifold.

9. Allow the container to fill with liquid by the force of gravity

(10–20 mL is recommended).

10. Close the clamps at the sample manifold inlet.

11. Close the clamps nearest the sample container.

12. Remove the filled manifold from the S.U.B. by welding a new
manifold onto the sample line, which will be used for taking the next
sample.

3.6.16 Dispense and harvest

1. Connect the bottom drain tubing set to the intended transfer line.

2. Open the clamp positioned at the bottom drain port.

3. Begin to drain, using a peristaltic pump.

4. Stop the impeller motor when volume reaches 20% maximum

volume.

5. Remove and store the drive shaft by reversing the steps used
during assembly.

6. Disable the temperature control to ensure that the S.U.B. does not
overheat.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 128

Chapter 3 | Operating information

7. When approximately three to five liters remain in the BPC, lift the
BPC at the top hanging tabs located opposite of the bottom drain
(this will pool media toward the drain).

8. Hold the bottom drain line near the floor while lifting the exhaust
filter side of the BPC to facilitate draining the final liter of harvest
media.

3.6.17 BPC disposal

After the drive shaft has been removed and the BPC has been drained,
the BPC can be removed from the outer support container. Filters
can be removed and integrity tested as needed according to the
user’s standard procedures. All product contact materials related to
the S.U.B. can be disposed of in an appropriate waste container or
incinerator. Note: For 2,000 L systems, do not lower the BPC lift until
the used bag has been removed from the tank.

3.6.18 S.U.B. shutdown

1. After the run is complete, verify that the motor agitation is off and
turn off the power to the outer support container by switching off
the main power disconnect.

2. If the S.U.B. hardware has come in contact with caustic materials

during the course of a run, rinse the affected areas with a light
water rinse, followed by normal routine cleaning (see section 3.6.19).

3. Loose items such as the drive shaft, tools, and RTD probes
should be returned to their storage locations to prevent accidental
damage.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 129

Chapter 3 | Operating information

3.6.19 Preparation for the next run

Between runs, the S.U.B. hardware (outer support container, probe

shelf, drive shaft, mixer drive, etc.) can be wiped down with a sanitary
wipe. The outer support container can be cleaned with standard
stainless steel cleaner. Store the drive shaft in the storage holder
located near the handle of the outer support container.

The S.U.B. hardware system can be cleaned to the extent of

standard laboratory cleaning procedures. Care should be taken to
ensure electrical connections have been disconnected and electrical
enclosures are closed tightly. It is also recommended that excess water
is not introduced under the heat shield or over the control panel. A
wipe-down with normal disinfectant solutions is sufficient. Avoid using
excessive amounts of liquid. The unit must be allowed to fully dry prior
to being brought back into operation.

3.7 Verification procedures

3.7.1 Mixing speed verification

To verify the mixing speed, use a calibrated tachometer. Expect

accuracy of ± 1.5 rpm or 1% of the setpoint, whichever is greater.
Speed scaling can be modified if the calibration needs to be adjusted.

3.7.2 Temperature controller verification

To verify the temperature controller/RTD, use a S.U.B. silicone

thermowell, the existing 3.175 mm (1/8 in.) outer diameter (OD) RTD and
a user-supplied calibrated temperature bath.

3.7.3 Pressure monitor verification (when present)

To verify the calibration of the pressure monitor, use a calibrated

pressure standard. Pressures can be verified by clamping the BPC inlet
line and supplying gas through the overlay gas filter. Expect accuracy
of ± 0.1 psi. The monitor can be calibrated manually by referencing
the monitor operator’s manual supplied in the Equipment Turnover
Package (ETP).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 130

Chapter 3 | Operating information

3.7.4 Load cell verification

It is recommended that the load cell manufacturer or a qualified

technician verify the load cells onsite. Expect an accuracy of ± 0.5 kg.
Basic load cell default parameters are listed in the electrical schematic
included with the ETP.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 131

 System features and
specifications
4
Chapter contents
4.1 Hardware features
4.2 Hardware specifications
4.3 Electrical control panel features
4.4 BPC specifications
4.5 Additional system component part numbers

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 132

Chapter 4 | System features and specifications

4.1 Hardware features

4.1.1 Design features for 50–250 L systems

Figures 4.1 and 4.2 (below) illustrate the hardware features of 50–250 L
S.U.B. systems. Electrical control panel (E-Box) features are illustrated
in section 4.3.

2 12

4
3
13
5
6
14

7 15

8
19
16
9
11 17 20
18

10

Figure 4.1. Front/side view of 50 L S.U.B. Figure 4.2. Back view of 50 L S.U.B.

1. Exhaust vent filter holder (optional) 12. Standard tool set: 10 mm (3/8 in.) x 16.9 Nm (150 in-lb.)
2. Mixing assembly with shield square torque wrench, load cell and motor cap lockout
3. Mixer motor wrench
4. Bearing port receiver with clamp 13. Cable management system
5. Liquid sight windows 14. Drive shaft, stored
6. Load cell display 15. Stainless steel (304) outer support container
7. Electrical control panel (E-Box), optional 16. Bleed valve
8. Probe hanger bracket 17. 0.95 cm (3/8 in.) Dimpled jacket (side)
9. Probe access windows 18. Cart assembly
10. Leveling casters 19. Bottom cutouts/pins for BPC attachment/alignment
11. Load cells 20. Quick-connect water inlet/outlet ports

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 133

Chapter 4 | System features and specifications

4.1.2 Design features for 500 L systems

Figures 4.3 and 4.4 (below) illustrate the features of 500 L S.U.B.
systems. Electrical control panel (E-Box) features are illustrated in
section 4.3.

12

2
4 13
3
5
14
6

16 15
8

11
17 18
9

19
10 20

Figure 4.3. Front/side view of 500 L S.U.B. Figure 4.4. Back view of 500 L S.U.B.

1. Exhaust vent filter holder 12. Standard tool set: 10 mm (3/8 in.) x 16.9 Nm
2. Mixing assembly with shield (150 in-lb.) square torque wrench, load cell and
3. Mixer motor motor cap lockout wrench
4. Bearing port receiver with clamp 13. Cable management system
5. Liquid sight windows 14. Drive shaft (2-piece), stored
6. Load cell display 15. Stainless steel (304) outer support container
7. Electrical control panel (E-Box), optional 16. 0.95 cm (3/8 in.) Dimpled jacket
8. Probe hanger bracket 17. Bleed valve
9. Probe access windows 18. Bottom cutouts/pins for BPC attachment/
10. Leveling casters alignment
11. Load cells (3) 19. Cart assembly
20. Quick-connect water inlet/outlet ports

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 134

Chapter 4 | System features and specifications

4.1.3 Design features for 1,000 L systems

Figures 4.5 and 4.6 (below) illustrate the features of 1,000 L S.U.B.
systems. Electrical control panel (E-Box) features are illustrated in
section 4.3.

1
8
17

13
9
2

14

18
4

10
5
19
11
15
12
6 20

16
Figure 4.5. Front/side view of 1,000 L S.U.B. Figure 4.6. Back view of 1,000 L S.U.B.

1. Mixing assembly with motor safety shield 13. 0.95 cm (3/8 in.) Dimpled jacket
2. Handheld controller (on bracket) 14. Stainless steel (304) outer support container
3. Drive shaft, stored (3-piece shown) 15. Bleed valve
4. BPC loading door and liquid sight windows 16. Quick-connect water inlet/outlet ports
5. Probe hanger bracket (with probe clips) 17. Standard tool set: 10 mm (3/8 in.) x 16.9 Nm (150 in-lb.)
6. Probe access windows square torque wrench, load cell and motor cap lockout
7. Exhaust vent filter holder wrench
8. Motor lift 18. Cable management system (optional)
9. Mixer motor 19. Bottom cutouts/pins for BPC attachment and alignment
10. Electrical control panel (E-Box), optional 20. Leveling casters
11. Load cell (3)
12. Cart assembly

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 135

Chapter 4 | System features and specifications

4.1.4 Design features for 2,000 L systems

Figures 4.7 and 4.8 (below) illustrate the features of 2,000 L S.U.B.
systems. Electrical control panel (E-Box) features are illustrated in
section 4.3.

1 10 11

11 19
3 15

4
12

6 16

7
20

13
8

9 14 21

17
22

18
Figure 4.7. Front/side view of 2,000 L S.U.B. Figure 4.8. Back view of 2,000 L S.U.B.

1. Motor lift 12. Standard tool set: 10 mm (3/8 in.) x 16.9 Nm (150 in-lb.)
2. Mixer motor square torque wrench, load cell and motor cap lockout
3. Mixing assembly with motor safety shield wrench
4. Handheld controller for motor adjustment (on bracket) 13. Load cell display
5. Emergency stop button (E-Stop) 14. Electrical control panel (E-Box), optional
6. Drive shaft, stored (4-piece shown) 15. 0.95 cm (3/8 in.) Dimpled jacket
7. Front access door and liquid sight windows 16. BPC lift control
8. Probe access window 17. Bleed valve
9. Probe hanger bracket (with probe clips) 18. Quick-connect water inlet/outlet ports
10. Exhaust vent filter holder 19. Stainless steel (304) outer support container
11. BPC lift (2) 20. Rear access door
21. Bottom cutouts/pins for BPC attachment and alignment
22. Load cell (3)

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 136

Chapter 4 | System features and specifications

4.2 Hardware specifications

The following tables and figures provide specifications for 50, 100,
250, 500, 1,000, and 2,000 L S.U.B.s. See section 1.2.3 for drive shaft
specifications.

Table 4.1. 50 L S.U.B. specifications.

AC and DC motors
Rated liquid working volume 50 L
Minimum liquid working volume 10 L
Bioreactor geometry

Total reactor volume (liquid & gas) 65.5 L

BPC chamber diameter 34.9 cm (13.75 in.)
BPC chamber shoulder height 80 cm (31.5 in.)
Liquid height at rated working volume 52.1 cm (20.5 in.)
Fluid geometry at working volume (height/diameter) ratio 1.5:1
Overall reactor geometry (height/diameter ratio) 1.9:1
Tank baffles No
Impeller (quantity x blade count) 1x3
Impeller scaling (impeller diameter/tank diameter) 1/3
Impeller

Impeller blade pitch (angle) 45°

Impeller diameter 11.11 cm (4.37 in.)
Impeller—calculated power number (N) 2.1
30–200 ± 1.5 rpm or 1% of setpoint,
Agitation speed range
whichever is greater
3
Nominal agitation rating—power/volume ratio 20 W/m
Nominal agitation—20% working volume 107 rpm
Nominal agitation—50% working volume 145 rpm
Nominal agitation—100% working volume 183 rpm
Agitation

Nominal tip speed 103.9 cm/s (204.6 ft./min.)

Counterclockwise mixing flow direction Down-pumping
Agitation shaft resolved angle 16.5°
Agitation shaft centerline offset 1.9 cm (0.75 in.)
Overall drive shaft length 85.09 cm (33.5 in.)
Drive shaft diameter 1.27 cm (0.5 in.)
Drive shaft poly-sheath outside diameter 2.54 cm (1 in.)
Impeller clearance from tank bottom 4.83 cm (1.9 in.)

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 137

Chapter 4 | System features and specifications

Table 4.2. 50 L S.U.B. specifications (continued).

AC motor DC motor
Agitation motor drive (type, voltage, phase) AC motor
Induction, 208 VAC, 3 N/A
only
Agitation motor drive (type, voltage) DC motor only N/A Brushless, 48 VDC
Motor power rating (AC motor) 186.4 W (0.25 hp) N/A
Motor power rating (DC motor) N/A 200 W (0.268 hp)
Motor
Motor torque rating 9.5 Nm (82 in-lb.) N/A
Gear reduction 10:1 N/A
Programmable VFD, remote panel interface, power
Standard N/A
fault auto restart
Motor communication methods 0-10 V, 4-20 mA,
N/A
(for external controller) Modbus
2 2 2 2
Jacket area: full/half volume 0.41 m (4.4 ft. ) / 0.19 m (2 ft. )
Jacket volume 2.4 L
Temperature control

Jacket flow rate at 3.4 bar (50 psi) 136 L/min.

Process connection 1.5 in. Sanitary tri-clamp
Nominal heating/cooling load (W) 500 W
Approximate liquid heat-up time (5°C to 37°C)—20%
1 hr
volume
Approximate liquid heat-up time (5°C to 37°C)—
1.1 hr
100% volume
RTD or thermocouple, 3.18 mm (1/8 in.) OD RTD: Pt-100 (standard)
116.4 cm (45.8 in.)
Overall width 64.21 cm (25.1 in.)
Support container

with E-Box
95.52 cm (38.0 in.)
Overall length 85.85 cm (33.8 in.)
with E-Box
Overall height (without filter) 141.8 cm (55.8 in.)
Dry skid weight (mass) 169.0 kg (372.5 lb.)
Wet skid weight—rated working volume (mass) 218.9 kg (482.7 lb.)
Ceiling height required for drive shaft loading 208.53 cm (82.1 in.)
Electrical power supply requirement 120/240 VAC, single, Dependent on
General

(voltage, phase, amp) 20/10 A controller

pH & DO probe—autoclavable type 12 mm diameter x 215–235 mm insertion
(Applisens, Broadley James, Mettler Toledo) length x 13.5 PG (pipe) thread
Noise level < 70 dB at 1.5 m
Operating temperature range Ambient to 40°C ± 0.5°C (104°F ± 0.9°F)
Recommended

parameters
operating

Motor speed 30–200 rpm

Volume range 10–50 L
Maximum bag pressure 0.03 bar (0.5 psi)

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 138

Chapter 4 | System features and specifications

Figure 4.9. Dimensions of 50 L S.U.B. (front view).

Figure 4.10. Dimensions of 50 L S.U.B. (top view).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 139

Chapter 4 | System features and specifications

Table 4.3. 100 L S.U.B. specifications.

AC motor DC motor
Rated liquid working volume 100 L
Minimum liquid working volume 20 L

Bioreactor geometry
Total reactor volume (liquid & gas) 120 L
BPC chamber diameter 43.8 cm (17.25 in.)
BPC chamber shoulder height 95.3 cm (37.5 in.)
Liquid height at rated working volume 66 cm (26 in.)
Fluid geometry at working volume (height/diameter) ratio 1.5:1
Overall reactor geometry (height/diameter ratio) 1.9:1
Tank baffles No
Impeller (quantity x blade count) 1x3
Impeller scaling (impeller diameter/tank diameter) 1/3
Impeller

Impeller blade pitch (angle) 45°

Impeller diameter 14.6 cm (5.75 in.)
Impeller—calculated power number (N) 2.1
30–200 ± 1.5 rpm or 1% of setpoint,
Agitation speed range
whichever is greater
3
Nominal agitation rating—power/volume ratio 20 W/m
Nominal agitation—20% working volume 85 rpm
Nominal agitation—50% working volume 116 rpm
Nominal agitation—100% working volume 146 rpm
Agitation

Nominal tip speed 110.7 cm/s (218 ft./min.)

Counterclockwise mixing flow direction Down-pumping
Agitation shaft resolved angle 16.5°
Agitation shaft centerline offset 2.54 cm (1 in.)
Overall drive shaft length 100.58 cm (39.6 in.)
Drive shaft diameter 1.27 cm (0.5 in.)
Drive shaft poly-sheath outside diameter 2.54 cm (1 in.)
Impeller clearance from tank bottom 5.08 cm (2 in.)

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 140

Chapter 4 | System features and specifications

Table 4.4. 100 L S.U.B. specifications (continued).

AC motor DC motor
Agitation motor drive (type, voltage, phase) AC motor only Induction, 208 VAC, 3 N/A
Agitation motor drive (type, voltage) DC motor only N/A Brushless, 48 VDC
Motor power rating (AC motor) 186.4 W (0.25 hp) N/A
Motor power rating (DC motor) N/A 200 W (0.268 hp)
Motor Motor torque rating 9.5 Nm (82 in-lb.) N/A
Gear reduction 10:1 N/A
Programmable VFD, remote panel interface, power fault
Standard N/A
auto restart
0–10 V, 4–20 mA,
Motor communication methods (for external controller) N/A
Modbus
Jacket area: full/half volume 0.60 m2 (6.5 ft.2) / 0.21 m2 (2.3 ft.2)
Jacket volume 4.5 L
Temperature control

Jacket flow rate at 3.4 bar (50 psi) 136 L/min.

Process connection 1.5 in. Sanitary tri-clamp
Nominal heating/cooling load 1,000 W
Approximate liquid heat-up time (5°C to 37°C)—20%
0.9 hr
volume
Approximate liquid heat-up time (5°C to 37°C)—100%
1.6 hr
volume
RTD or thermocouple, 3.18 mm (1/8 in.) OD RTD: Pt-100 (standard)
109.2 cm (43.0 in.)
Overall width 61.6 cm (24.3 in.)
with E-box
100.2 cm (39.4 in.)
container
Support

Overall length 93.24 cm (36.71 in.)

with E-box
Overall height (without filter bracket) 144.2 cm (56.8 in.)
Dry skid weight (mass) 199.8 kg (440.5 lb.)
Wet skid weight—rated working volume (mass) 299.8 kg (661.0 lb.)
Ceiling height required for drive shaft loading 232.66 cm (91.6 in.)
Electrical power supply requirement 120/240 VAC, Single, Dependent on
General

(voltage, phase, amp) 20/10 A controller

pH & DO probe—autoclavable type 12 mm diameter x 215–235 mm insertion
(Applisens, Broadley James, Mettler Toledo) length x 13.5 PG (pipe) thread
Noise level < 70 dB at 1.5 m
Operating temperature range Ambient to 40°C ± 0.5°C (104°F ± 0.9°F)
Recommended

parameters
operating

Motor speed 30–200 rpm

Volume range 20–100 L
Maximum bag pressure 0.03 bar (0.5 psi)

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 141

Chapter 4 | System features and specifications

Figure 4.11. Dimensions of 100 L S.U.B. (front view).

Figure 4.12. Dimensions of 100 L S.U.B. (top view).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 142

Chapter 4 | System features and specifications

Table 4.5. 250 L S.U.B. specifications.

AC and DC motors
Rated liquid working volume 250 L
Minimum liquid working volume 50 L

Bioreactor geometry
Total reactor volume (liquid & gas) 316 L
BPC chamber diameter 59.7 cm (23.5 in.)
BPC chamber shoulder height 115.6 cm (45.5 in.)
Liquid height at rated working volume 91.4 cm (36 in.)
Fluid geometry at working volume (height/diameter) ratio 1.5:1
Overall reactor geometry (height/diameter ratio) 1.9:1
Tank baffles No
Impeller (quantity x blade count) 1x3
Impeller scaling (impeller diameter/tank diameter) 1/3
Impeller

Impeller blade pitch (angle) 45°

Impeller diameter 20 cm (7.875 in.)
Impeller—calculated power number (N) 2.1
30–150 ± 1.5 rpm or 1% of setpoint,
Agitation speed range
whichever is greater
3
Nominal agitation rating—power/volume ratio 20 W/m
Nominal agitation—20% working volume 69 rpm
Nominal agitation—50% working volume 93 rpm
Nominal agitation—100% working volume 117 rpm
Agitation

Nominal tip speed 123.6 cm/s (243.3 ft./min.)

Counterclockwise mixing flow direction Down-pumping
Agitation shaft resolved angle 16.5°
Agitation shaft centerline offset 3.3 cm (1.3 in.)
Overall drive shaft length 120.90 cm (47.6 in.)
Drive shaft diameter 1.27 cm (0.5 in.)
Drive shaft poly-sheath outside diameter 2.54 cm (1 in.)
Impeller clearance from tank bottom 6.91 cm (2.72 in.)

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 143

Chapter 4 | System features and specifications

Table 4.6. 250 L S.U.B. specifications (continued).

AC motor DC motor
Agitation motor drive (type, voltage, phase) AC motor only Induction, 208 VAC, 3 N/A
Agitation motor drive (type, voltage) DC motor only N/A Brushless, 48 VDC
Motor power rating (AC motor) 186.4 W (0.25 hp) N/A
Motor power rating (DC motor) N/A 400 W (0.536 hp)
Motor Motor torque rating 11.5 Nm (102 in-lb.) N/A
Gear reduction 12.5:1 N/A
Programmable VFD, remote panel interface, power faults
Standard N/A
auto restart
0–10 V, 4–20 mA,
Motor communication methods (for external controller) N/A
Modbus
2 2
Jacket area: full/half volume 1.26 m (13.6 ft. ) /
2 2
0.54 m (5.8 ft. )
Jacket volume 8.6 L
Temperature control

Jacket flow rate at 3.4 bar (50 psi) 136 L/min.

Process connection 1.5 in. Sanitary tri-clamp
Nominal heating/cooling load (W) 2,500 W
Approximate liquid heat-up time (5°C to 37°C)—20%
1.1 hr
volume
Approximate liquid heat-up time (5°C to 37°C)—100%
3.4 hr
volume
RTD or thermocouple, 3.18 mm (1/8 in.) OD RTD: Pt-100 (standard)
133 cm (52 in.)
Overall width 97.9 cm (38 in.)
Support container

with E-box
Overall length 108.9 cm (43 in.)
Overall height (without filter bracket) 159.5 cm (63 in.)
Dry skid weight (mass) 223.6 kg (493 lb.)
Wet skid weight—rated working volume (mass) 473.6 kg (1044 lb.)
Ceiling height required for
267.46 cm (105.3 in.)
drive shaft loading
Electrical power supply requirement 120/240 VAC, single, Dependent on
General

(voltage, phase, amp) 20/10 A controller

pH & DO probe–autoclavable type 12 mm diameter x 215–235 mm insertion
(Applisens, Broadley James, Mettler Toledo) length x 13.5 PG thread
Noise level < 70 dB at 1.5 m
Operating temperature range Ambient to 40°C ± 0.5°C (104°F ± 0.9°F)
Recommended

parameters
operating

Motor speed 30–150 rpm

Volume range 50–250 L
Maximum bag pressure 0.03 bar (0.5 psi)

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 144

Chapter 4 | System features and specifications

Figure 4.13. Dimensions of 250 L S.U.B. (front view).

Figure 4.14. Dimensions of 250 L S.U.B. (top view).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 145

Chapter 4 | System features and specifications

Table 4.7. 500 L S.U.B. specifications.

AC and DC motors
Rated liquid working volume 500 L
Minimum liquid working volume 100 L

Bioreactor geometry
Total bioreactor volume (liquid & gas) 660 L
BPC chamber diameter 75.56 cm (29.75 in.)
BPC chamber shoulder height 152.4 cm (60 in.)
Liquid height at rated working volume 113.36 cm (44.63 in.)
Fluid geometry at working volume (height/diameter) ratio 1.5:1
Overall bioreactor geometry (height/diameter) ratio 1.9:1
Tank baffles No
Impeller (quantity x blade count) 1x3
Impeller scaling (impeller diameter/tank diameter) 1/3
Impeller

Impeller blade pitch 45°

Impeller diameter 25.1 cm (9.875 in.)
Impeller—calculated power number (N) 2.1
30–150 ± 1.5 rpm or 1% of setpoint,
Agitation speed range
whichever is greater
Nominal agitation rating (power/volume ratio) 20 W/m3
Nominal agitation—20% working volume 59 rpm
Nominal agitation—50% working volume 80 rpm
Nominal agitation—100% working volume 101 rpm
Agitation

Nominal tip speed 137.2 cm/s (270 ft./min.)

Counterclockwise mixing flow direction Down-pumping
Agitation shaft resolved angle 16.5°
Agitation shaft centerline offset 5.08 cm (2 in.)
Overall drive shaft length 155.7 cm (61.3 in.)
Drive shaft diameter 1.90 cm (0.75 in.)
Drive shaft poly-sheath outside diameter 2.54 cm (1 in.)
Impeller clearance from tank bottom 7.62 cm (3 in.)

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 146

Chapter 4 | System features and specifications

Table 4.8. 500 L S.U.B. specifications (continued).

AC motor DC motor
Agitation motor drive (type, voltage, phase) AC motor only Induction, 208 VAC, 3 N/A
Agitation motor drive (type, voltage) DC motor only N/A Brushless, 48 VDC
Motor power rating (AC motor) 372.8 W (0.5 hp) N/A
Motor power rating (DC motor) N/A 400 W (0.536 hp)
Motor
Motor torque rating 9.5 Nm (82 in-lb.) N/A
Gear reduction 10:1 N/A
Programmable VFD, remote panel interface, power fault
Standard N/A
auto restart
0–10 V; 4–20 mA;
Motor communication methods (for external controller) N/A
Modbus
2 2
Jacket area: full/half volume 1.99 m (21.4 ft. ) /
2 2
0.78 m (8.4 ft. )
Jacket volume 15.2L
Temperature control

Jacket flow rate at 3.4 bar (50 psi) 136 L/min.

Process connection 1.5 in. Sanitary tri-clamp
Nominal heating/cooling load (W) 5,000 W
Approximate liquid heat-up time (5°C to 37°C)—20%
1.1 hr
volume
Approximate liquid heat-up time (5°C to 37°C)—100%
2.2 hr
volume
RTD or thermocouple, 3.18 mm (1/8 in.) OD RTD: Pt-100 (standard)
148.3 cm (58 in.)
Overall width 110.6 cm (43 in.)
Support container

with E-Box
Overall length 124.8 cm (49 in.)
Overall height (without filter bracket) 195.9 cm (77 in.)
Dry skid weight (mass) 353.8 kg (780 lb.)
Wet skid weight—rated working volume (mass) 853.8 kg (1882.3 lb.)
Ceiling height required for drive shaft loading 282.19 cm (111.1 in.)
120/240 VAC, Dependent on
Electrical power supply requirement (voltage, phase, amp)
General

single, 20/10 A controller

pH & DO probe—autoclavable type 12 mm diameter x 215–235 mm insertion
(Applisens, Broadley James, Mettler Toledo) length x 13.5 PG (pipe) thread
Noise level < 70 dB at 1.5 m
Operating temperature range Ambient to 40°C ± 0.5°C (104°F ± 0.9°F)
Recommended

parameters
operating

Motor speed 30–150 rpm

Volume range 100–500 L
Maximum BPC pressure 0.03 bar (0.5 psi)

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 147

Chapter 4 | System features and specifications

Figure 4.15. Dimensions of 500 L S.U.B. (front view).

Figure 4.16. Dimensions of 500 L S.U.B. (top view).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 148

Chapter 4 | System features and specifications

Table 4.9. 1,000 L S.U.B. specifications.

AC and DC motors
Rated liquid working volume 1,000 L
Minimum liquid working volume for 5:1 operation 200 L
Total bioreactor volume (liquid & gas) 1,320 L

Bioreactor geometry
BPC chamber diameter 95.9 cm (37.75 in.)
BPC chamber shoulder height 200.7 cm (79 in.)
Liquid height at rated working volume 142.2 cm (56 in.)
Fluid geometry at working volume
1.5:1
(height/diameter) ratio
Overall bioreactor geometry
1.9:1
(height/diameter) ratio
Tank baffles No
Impeller (quantity x blade count) 1x3
Impeller scaling (impeller diameter/tank diameter) 1/3
Impeller

Impeller blade pitch 45°

Impeller diameter 32.1 cm (12.625 in.)
Impeller—calculated power number (N) 2.1
20–110 ± 1.5 rpm or 1% of setpoint,
Agitation speed range for 5:1 operation
whichever is greater
3
Nominal agitation rating (power/volume ratio) 20 W/m
Nominal agitation—20% working volume (5:1 operation
50 rpm
only)
Nominal agitation—50% working volume 68 rpm
Nominal agitation—100% working volume 86 rpm
Agitation

Nominal tip speed 146.1 cm/s (287.6 ft./min.)

Counterclockwise mixing flow direction Down-pumping
Agitation shaft resolved angle for 5:1 operation 15.3°
Agitation shaft centerline offset 5.08 cm (2.0 in.)
Overall drive shaft length 167.6 cm (66 in.)
Drive shaft diameter 1.90 cm (0.75 in.)
Drive shaft poly-sheath outside diameter 3.49 cm (1.375 in.)
Impeller clearance from tank bottom for 5:1 systems 9.73 cm (3.83 in.)
Power supply requirements 24 VDC, 90 psi of air
Motor
lift

Weight 115.21 kg (254 lb.)

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 149

Chapter 4 | System features and specifications

Table 4.10. 1,000 L S.U.B. specifications (continued).

AC and DC motors
Agitation motor drive (type, voltage, phase) Induction, 208 AC, 3
Motor power rating 372.8 W (0.5 hp)
Motor torque rating 27.7 Nm (245 in-lb.)

Motor
Gear reduction 15:1
Programmable VFD, remote panel interface, power fault
Standard
auto restart
Motor communication methods (for external controller) 0–10 V; 4–20 mA; Modbus
2 2
Jacket area: full/half volume 3.31 m (35.6 ft. ) /
2 2
1.38 m (14.9 ft. )
Jacket volume 23.5 L
Temperature control

Jacket flow rate at 3.4 bar (50 psi) 136 L/min.

Process connection 1.5 in. Sanitary tri-clamp
Nominal heating/cooling load (W) 9,000 W
Approximate liquid heat-up time (5°C to 37°C)—20%
1.2 hr
volume
Approximate liquid heat-up time (5°C to 37°C)—100%
4.1 hr
volume
RTD or thermocouple, 3.18 mm (1/8 in.) OD RTD: Pt-100 (standard)
Overall width 167.1 cm (65.8 in.) with E-Box
Overall length 136.6 cm (53.8 in.) with E-Box
container
Support

Overall height (without filter bracket) 249.4 cm (98.2 in.)

Dry skid weight (mass) 655.01 kg (1,444 lb.)
Wet skid weight—rated working volume (mass) 1,655.01 kg (3,649 lb.)
Ceiling height required for drive shaft loading 287 cm (113 in.)
Electrical power supply requirement (voltage, phase, 208–240 VAC, single, 30 A/dependent on
General

amp) controller
pH & DO probe—autoclavable type 12 mm diameter x 215–235 mm insertion
(Applisens, Broadley James, Mettler Toledo) length x 13.5 PG (pipe) thread
Noise level < 70 dB at 1.5 m
Operating temperature range Ambient to 40°C ± 0.5°C (104°F ± 0.9°F)
Recommended

parameters
operating

Motor speed 20–110 rpm

Volume range 200–1,000 L
Maximum BPC pressure 0.03 bar (0.5 psi)

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 150

Chapter 4 | System features and specifications

Figure 4.17. Dimensions of 1,000 L S.U.B. (front view).

Figure 4.18. Dimensions of 1,000 L S.U.B. (top view).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 151

Chapter 4 | System features and specifications

Table 4.11. 2,000 L S.U.B. specifications.

AC and DC motors
Rated liquid working volume 2,000 L
Minimum liquid working volume for 5:1 operation 400 L
Total bioreactor volume (liquid & gas) 2,575 L

Bioreactor geometry
BPC chamber diameter 119.4 cm (47 in.)
BPC chamber shoulder height 229.9 cm (90.5 in.)
Liquid height at rated working volume 178.7 cm (70.35 in.)
Fluid geometry at working volume
1.5:1
(height/diameter) ratio
Overall bioreactor geometry
1.9:1
(height/diameter) ratio
Tank baffles No
Impeller (quantity x blade count) 1x3
Impeller scaling (impeller diameter/tank diameter) 1/3
Impeller

Impeller blade pitch 45°

Impeller diameter 39.7 cm (15.67 in.)
Impeller—calculated power number (N) 2.1
Standard operation: 20–75* ± 1.5 rpm or 1%
Agitation speed range for 5:1 operation
of setpoint, whichever is greater
3
Nominal agitation rating (power/volume ratio) 20 W/m
Nominal agitation—20% working volume (5:1 operation
44 rpm
only)
Nominal agitation—50% working volume 60 rpm
Nominal agitation—100% working volume 75 rpm
Agitation

Minimum acceleration and deceleration rate 60 seconds

Nominal tip speed 154.9 cm/s (305 ft./min.)
Counterclockwise mixing flow direction Down-pumping
Agitation shaft resolved angle for 5:1 operation 16.6°
Agitation shaft centerline offset 6.7 cm (2.63 in.)
Overall drive shaft length 215.6 cm (84.9 in.)
Drive shaft diameter 1.91 cm (0.75 in.)
Drive shaft poly-sheath outside diameter 3.51 cm (1.38 in.)
Impeller clearance from tank bottom for 5:1 systems 13.9 cm (5.49 in.)
Ceiling height required for 2-piece drive shaft loading 381 cm (150 in.)
Ceiling height required for 4-piece drive shaft loading 353.1 cm (139 in.)
208–240 VAC, single, 30 A/dependent on
General

Electrical power supply requirement (voltage, phase, amp)

controller
pH & DO probe—autoclavable type 12 mm diameter x 215–235 mm insertion
(Applisens, Broadley James, Mettler Toledo) length x 13.5 PG (pipe) thread
Noise level < 70 dB at 1.5 m

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 152

Chapter 4 | System features and specifications

Table 4.12. 2,000 L S.U.B. specifications (continued).

AC and DC motors
Agitation motor drive (type, voltage, phase) Induction, 208 AC, 3
Motor power rating 372.8 W (0.5 hp)
Motor torque rating 27.7 Nm (245 in-lb.)
Motor
Gear reduction 15:1
Programmable VFD, remote panel interface, power fault
Standard
auto restart
Motor communication methods (for external controller) 0–10 V; 4–20 mA; Modbus
2 2
4.54 m (48.9 ft. ) /
Jacket area: full/half volume 2 2
2.47 m (26.6 ft. )
Jacket volume 44 L
Temperature control

Jacket flow rate at 3.4 bar (50 psi) 75 L/min.

Process connection 1.5 in. Sanitary tri-clamp
Nominal heating/cooling load (W) 18,000 W
Approximate liquid heat-up time (5°C to 37°C)—20%
1.4 hr
volume
Approximate liquid heat-up time (5°C to 37°C)—100%
4 hr
volume
RTD or thermocouple, 3.18 mm (1/8 in.) OD RTD: Pt-100 (standard)
Overall width 163 cm (64.2 in.) with E-Box
Overall length 157 cm (61.8 in.)
container
Support

Overall height (without filter bracket) 307.8 cm (121.2 in.)

Dry skid weight (mass) 962.1 kg (2,121 lb.)
Wet skid weight—rated working volume (mass) 2,962.1 kg (6,530 lb.)
Operating temperature range Ambient to 40°C ± 0.5°C (104°F ± 0.9°F)
Recommended

parameters
operating

Standard operation: 20–75* rpm, working

Motor speed
volumes must stay above 50% during agitation
Volume range 400–2,000 L
Maximum BPC pressure 0.03 bar (0.5 psi)
BPC lift

Electrical power supply requirement 120–240 VAC, 50/60 Hz, 10 A

Duty cycle for motor Tested to 5 cycles/10 minutes
Power supply requirements 24 VDC, 90 psi of air
Motor
lift

Weight 126.1 kg (278 lb.)

* WARNING: Mixing speeds must stay within the recommended operating parameters. Higher speed operation compromises
system reliability and will void standard Thermo Fisher warranties. Your control strategy should include governors that regulate
rpm based on liquid volume as well as safety interlocks that disable mixing when the liquid drops below the recommended
volume. See section 3.6.5 for more information about operating parameters, agitation speed governors, and safety interlocks.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 153

Chapter 4 | System features and specifications

Figure 4.19. Dimensions of 2,000 L S.U.B. (front view).

Figure 4.20. Dimensions of 2,000 L S.U.B. (top view).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 154

Chapter 4 | System features and specifications

4.3 E-Box features

Figure 4.21 illustrates the features of the E-Box available for 50, 100,
250, 500, 1,000, and 2,000 L S.U.B. units with AC motors. Figure 4.22
illustrates the bottom view of the E-Box.

Pressure
display
Power reset
switch

Temperature
Motor controller display
power switch
Motor speed
control keypad

Emergency
stop button
Main switch

Figure 4.21. Front view of the E-Box for 50–2,000 L

S.U.B.s.

Buzzer
Spare 1/4 liquid
Continuous power: tight
For devices that
should not be RTD input signal
turned off during an
E-Stop. Examples Emergency shut-off power:
are recording For devices that should be
devices, sensors turned off when an operator
and controllers. activates an E-Stop. Examples
The maximum for are pumps, motors, and any
each receptacle is device that could harm an
1/2 amp. operator or ruin a batch during
Figure 4.22. Bottom view of all E-Boxes. an E-Stop. The maximum for
each receptacle is 2 amps.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 155

Chapter 4 | System features and specifications

4.4 BPC specifications

The following figures and tables provide specification information for
5:1 BPCs in 50, 100, 250, 500, 1,000, and 2,000 L sizes.

5:1 S.U.B. 50 L BPC with AseptiQuik connector ports

Specification information for the numbered items in Figure 4.23 is
located in Table 4.13 on the following page.

1
4–7
2
8

Impeller 3 9

Front face 14

10
11

12–13
15

Back face

Figure 4.23. Front and back faces of 50 L S.U.B. BPC.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 156

Chapter 4 | System features and specifications

Table 4.13. 50 L BPC specification information.

Item Description Tubing set (ID x OD x length) End treatment

6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 152 cm (60 in.) Reduced
1 Inoculum addition Plugged
to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm (12 in.)

Hydrophobic vent filter

Overlay gas with Emflon II, connected
2 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 15 cm (6 in.)
sparger to 15 cm (6 in.) C-Flex
tubing
6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 8 cm (3 in.) connected to
Cross flow Meissner Steridyne
3 check valve and 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 183 cm
sparger 50 mm (1.97 in.) filter
(72 in.)
CPC AseptiQuik aseptic
4–7 Probe ports (4) 12.7 mm (1/2 in.) tube ports
connectors
12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 152 cm (60 in.)
Reduced to 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm
Bottom drain Plugged and 9.5 mm
8 (12 in.) Splits to 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex Tubing x 30 cm
harvest (3/8 in.) MPC insert
(12 in.) Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm
(12 in.) and 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm (12 in.)
Luer lock and SterilEnz
Thermowell/small Thermowell adapter for 3.2 mm (1/8 in.) diameter RTD and 3.2 mm (1/8 in.) x
9 pouch with injection site
volume sample 6.4 mm (1/4 in.) C-Flex tubing x 46 cm (18 in.)
assembly

6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 15 cm (6 in.)

10 Base addition Plugged
Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 152 cm (60 in.)

Pressure sensor CPC AseptiQuik aseptic

11 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 8 cm (3 in.)
port connector
9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 152 cm (60 in.) SteriEnz pouch with
Splits to 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 30 cm (12 in.) injection site assembly
12–13 Feed lines
Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm (12 in.) and 9.5 mm (3/8 in.)
and 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm (12 in.) MPC body
CPC AseptiQuik
12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 20 cm (8 in.)
aseptic connector—
14 Exhaust line Connected to 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 15 cm (6
Pall Kleenpak 0.2 micron
in.) and 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 25 cm (10 in.)
exhaust vent filter

Drilled hole
Meissner Steridyne 0.2
sparger 8.9 cm 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 8 cm (3 in.)
micron hydrophobic filter
15 (3.5 in.) disk with Connected to check valve and 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex
connected to 15 cm
360 x 0.178 mm tubing x 97 cm (38 in.)
(6 in.) C-Flex
(0.007 in.) holes

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 157

Chapter 4 | System features and specifications

5:1 S.U.B. 100 L BPC with AseptiQuik connector ports

Specification information for the numbered items in Figure 4.24 is
located in Table 4.14 on the following page.

3 4–7
2
8

Impeller 9

Front face
14
10

11

15
12–13

Back face

Figure 4.24. Front and back faces of 100 L S.U.B. BPC.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 158

Chapter 4 | System features and specifications

Table 4.14. 100 L BPC specification information.

Item Description Tubing set (ID x OD x length) End treatment

6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 152 cm (60 in.) Reduced
1 Inoculum addition Plugged
to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm (12 in.)
Hydrophobic vent
Overlay gas filter with Emflon II,
2 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 20 cm (8 in.)
sparger connected to 15 cm
(6 in.) C-Flex tubing
6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 8 cm (3 in.) connected to
Cross flow Meissner Steridyne
3 check valve and 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 183 cm
sparger 50 mm (1.97 in.) filter
(72 in.)
CPC AseptiQuik aseptic
4–7 Probe ports (4) 12.7 mm (1/2 in.) tube ports
connectors
12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 152 cm (60 in.)
Reduced to 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm
Bottom drain Plugged and 9.5 mm
8 (12 in.) Splits to 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 30 cm
harvest (3/8 in.) MPC insert
(12 in.) Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm
(12 in.) and 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm (12 in.)
Thermowell/small Thermowell adapter for 6.4 mm (1/4 in.) diameter RTD and SterilEnz pouch with
9
volume sample 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 46 cm (18 in.) injection site assembly

6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 15 cm (6 in.)

10 Base addition Plugged
Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 152 cm (60 in.)

Pressure sensor CPC AseptiQuik aseptic

11 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 8 cm (3 in.)
port connector
9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 152 cm (60 in.) SteriEnz pouch with
Splits to 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 30 cm (12 in.) injection site assembly
12–13 Feed lines
Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm (12 in.) and 9.5 mm (3/8 in.)
and 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm (12 in.) MPC body
CPC AseptiQuik
12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 20 cm (8 in.)
aseptic connector—
14 Exhaust line Connected to 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 15 cm
Pall Kleenpak 0.2 micron
(6 in.) and 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 25 cm (10 in.)
exhaust vent filter
Drilled hole
Meissner Steridyne 0.2
sparger 8.9 cm 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 8 cm (3 in.)
micron hydrophobic filter
15 (3.5 in.) disk with Connected to check valve and 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex
connected to 15 cm
570 x 0.178 mm tubing x 114 cm (45 in.)
(6 in.) C-Flex
(0.007 in.) holes

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 159

Chapter 4 | System features and specifications

5:1 S.U.B. 250 L BPC with AseptiQuik connector ports

Specification information for the numbered items in Figure 4.25 is
located in Table 4.15 on the following page.

Impeller 1–4

6 7
Front face

9
10 11

12

15

13–14

Back face

Figure 4.25. Front and back faces of 250 L S.U.B. BPC.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 160

Chapter 4 | System features and specifications

Table 4.15. 250 L BPC specification information.

Item Description Tubing set (ID x OD x length) End treatment
CPC AseptiQuik aseptic
1–4 Probe ports (4) 12.7 mm (1/2 in.) tube ports
connectors
12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 152 cm (60 in.)
Reduced to 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm
Bottom drain Plugged and 9.5 mm
5 (12 in.) Splits to 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 30 cm
harvest (3/8 in.) MPC insert
(12 in.) Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm
(12 in.) and 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm (12 in.)
6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 8 cm (3 in.) connected to
Cross flow Meissner Steridyne
6 check valve and 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 183 cm
sparger 50 mm (1.97 in.) filter
(72 in.)
Thermowell/small Thermowell adapter for 6.4 mm (1/4 in.) diameter RTD and SterilEnz pouch with
7
volume sample 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 46 cm (18 in.) injection site assembly
CPC AseptiQuik aseptic
19.1 mm (3/4 in.) x 25.4 mm (1 in.) C-Flex tubing x 30 cm (12 in.)
connector, (2) Meissner
8 Exhaust line Splits to 19.1 mm (3/4 in.) x 25.4 mm (1 in.) C-Flex tubing x 15 cm (6 in.) and
Ultracap 0.2 micron
19.1 mm (3/4 in.) x 25.4 mm (1 in.) C-Flex tubing x 15 cm (6 in.)
hydrophobic filters
6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 152 cm (60 in.) Reduced
9 Inoculum addition Plugged
to 3.2 mm (1/8 in.) x 6.4 mm 1/4 in.) C-Flex tubing x 30 cm (12 in.)
Pressure sensor CPC AseptiQuik aseptic
10 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 8 cm (3 in.)
port connector
Meissner Steridyne 0.2
Overlay gas micron hydrophobic filter
11 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 15 cm (6 in.)
sparger connected to 15 cm
(6 in.) C-Flex
6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 15 cm (6 in.)
12 Base addition Plugged
Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 152 cm (60 in.)
9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 152 cm (60 in.) SteriEnz pouch with
Splits to 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 30 cm (12 in.) injection site assembly
13–14 Feed lines
Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm (12 in.) and 9.5 mm (3/8 in.)
and 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm (12 in.) MPC body
Drilled hole
Meissner Steridyne 0.2
sparger 12.2 cm 6.4 mm (1/4 in.) ID x 11.1 mm (7/16 in.) C-Flex tubing x 8 cm (3 in.)
micron hydrophobic filter
15 (4.8 in.) disk with Connected to check valve and 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) OD C-Flex
connected to 15 cm
760 x 0.233 mm tubing x 150 cm (59 in.)
(6 in.) C-Flex
(0.009 in.) holes

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 161

Chapter 4 | System features and specifications

5:1 S.U.B. 500 L BPC with AseptiQuik connector ports

Specification information for the numbered items in Figure 4.26 is
located in Table 4.16 on the following page.

Impeller 1–5

8
7
Front face

11
12–13

10

15
14
16

Back face

Figure 4.26. Front and back faces of 500 L S.U.B. BPC.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 162

Chapter 4 | System features and specifications

Table 4.16. 500 L BPC specification information.

Item Description Tubing set (ID x OD x length) End treatment
CPC AseptiQuik aseptic
1–5 Probe ports (4) 12.7 mm (1/2 in.) tube ports
connectors
12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 152 cm (60 in.)
Reduced to 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm
Bottom drain Plugged and 9.5 mm
6 (12 in.) Splits to 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 30 cm
harvest (3/8 in.) MPC insert
(12 in.) Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm
(12 in.) and 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm (12 in.)
6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 8 cm (3 in.) connected to
Meissner Steridyne
7 Cross flow sparger check valve and 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 183 cm
50 mm (1.97 in.) filter
(72 in.)
Thermowell/small Thermowell adapter for 6.4 mm (1/4 in.) diameter RTD and SterilEnz pouch with
8
volume sample 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 46 cm (18 in.) injection site assembly
19.1 mm (3/4 in.) x 25.4 mm (1 in.) C-Flex tubing x 30 cm (12 in.) Meissner Ultracap 0.2
9 Exhaust line Splits to 19.1 mm (3/4 in.) x 25.4 mm (1 in.) C-Flex tubing x 15 cm (6 in.) and micron hydrophobic
19.1 mm (3/4 in.) x 25.4 mm (1 in.) C-Flex tubing x 15 cm (6 in.) filters
Hydrophobic vent
Overlay gas filter with Emflon II,
10 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 15 cm (6 in.)
sparger connected to 15 cm
(6 in.) C-Flex
Pressure sensor CPC AseptiQuik aseptic
11 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 8 cm (3 in.)
port connector
9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 213 cm (84 in.) SteriEnz pouch with
Splits to 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 30 cm (12 in.) injection site assembly
12–13 Feed lines
Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm (12 in.) and 9.5 mm (3/8 in.)
and 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm (12 in.) MPC body
6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 213 cm (84 in.)
14 Base addition Plugged
Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm (12 in.)
6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 15 cm (6 in.) Reduced to
15 Inoculum addition Plugged
3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 213 cm (84 in.)
Drilled hole
Kleenpak Emflon 0.2
sparger 17.14 cm 6.4 mm (1/4 in.) ID x 11.1 mm (7/16 in.) C-Flex tubing x 8 cm (3 in.)
micron hydrophobic
16 (6.75 in.) disk with Connected to check valve and 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) OD C-Flex
filter connected to 15
980 x 0.368 mm tubing x 97 cm (38 in.)
cm (6 in.) C-Flex
(0.014 in.) holes

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 163

Chapter 4 | System features and specifications

5:1 S.U.B. 1,000 L BPC with AseptiQuik connector ports

Specification information for the numbered items in Figure 4.27 is
located in Table 4.17 on the following page.

Impeller 1
2–6

8
Front face

11
10 12–13

14 16
15

Back face

Figure 4.27. Front and back faces of 1,000 L S.U.B. BPC.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 164

Chapter 4 | System features and specifications

Table 4.17. 1,000 L BPC specification information.

Item Description Tubing set (ID x OD x length) End treatment
6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 8 cm (3 in.) connected to
Meissner Steridyne
1 Cross flow sparger check valve and 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 183 cm
50 mm (1.97 in.) filter
(72 in.)
CPC AseptiQuik aseptic
2–6 Probe ports (4) 12.7 mm (1/2 in.) tube ports
connectors
12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 152 cm (60 in.)
Reduced to 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm
Bottom drain Plugged and 9.5 mm
7 (12 in.) Splits to 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 30 cm
harvest (3/8 in.) MPC insert
(12 in.) Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm
(12 in.) and 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm (12 in.)
Thermowell/small Thermowell adapter for 6.4 mm (1/4 in.) diameter RTD and SterilEnz pouch with
8
volume sample 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 60 cm (24 in.) injection site assembly
(2) Meissner Ultracap
12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 30 cm (12 in.) 0.2 micron hydrophobic
9 Exhaust line Splits to 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 15 cm (6 in.) filters connected to
and 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 15 cm (6 in.) 15 cm (6 in.) C-Flex
tubing
Kleenpak Emflon II
Overlay gas 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 10 cm (4 in.) Reduced to
10 capsule and pressure
sparger 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 213 cm (84 in.)
transducer
Pressure sensor CPC AseptiQuik aseptic
11 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 8 cm (3 in.)
port connector
9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 213 cm (84 in.) SteriEnz pouch with
Splits to 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 30 cm (12 in.) injection site assembly
12–13 Feed lines
Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm (12 in.) and 9.5 mm (3/8 in.)
and 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 30 cm (12 in.) MPC body
6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 15 cm (6 in.)
14 Base addition Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 213 cm Plugged
(84 in.)
6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 213 cm (84 in.) Reduced
15 Inoculum addition Plugged
to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm (12 in.)
Drilled hole
Meissner Steridyne 0.2
sparger 17.1 cm 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 8 cm (3 in.)
micron hydrophobic
16 (6.75 in.) disk with Connected to check valve and 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex
filter connected to
1,180 x 0.445 mm tubing x 264 cm (97 in.)
15 cm (6 in.) C-Flex
(0.018 in.) holes

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 165

Chapter 4 | System features and specifications

5:1 S.U.B. 2,000 L BPC with AseptiQuik connector ports

Specification information for the numbered items in Figure 4.28 is
located in Table 4.18 on the following page.

1 2–6
Impeller
9

13 11

Front face
8
14
10

16 17
18
19 12

21
Tri-clamp port

23 20

22

15
Back face

Figure 4.28. Front and back faces of 2,000 L S.U.B. BPC.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 166

Chapter 4 | System features and specifications

Table 4.18. 2,000 L BPC specification information.

Item Description Tubing set (ID x OD x length) End treatment
6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 8 cm (3 in.)
Meissner Steridyne
1 Cross flow sparger connected to check valve and 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.)
50 mm (1.97 in.) filter
C-Flex tubing x 183 cm (72 in.)
CPC AseptiQuik aseptic
2–6 Probe ports (5) 12.7 mm (1/2 in.) tube ports
connectors
19.1 mm (3/4 in.) x 25.4 mm (1 in.) C-Flex tubing x 213 cm (84 in.) splits
Media fill/auxiliary CPC AseptiQuik aseptic
7–8 to 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 61 cm (24 in.)
drain lines (2) connectors
and 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 61 cm (24 in.)
SterilEnz pouch with
12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 10 cm (4 in.)
injection site assembly
9–10 Feed lines (2) Splits to 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 25 cm (10 in.)
and 9.5 mm (3/8 in.)
and 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 25 cm (10 in.)
MPC body
Meissner Steridyne 0.2
micron hydrophobic
Drilled hole sparger
filter with 9.5 mm
17.1 cm (6.75 in.) 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 8 cm (3 in.)
(3/8 in.) C-Flex
11–12 disk with 1,380 Connected to check valve and 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex
tubing (Y-connector
(690 x 0.582 mm) tubing x 183 cm (72 in.)
and quick-connects
(0.023 in.) holes
provided for joining
lines)
Thermowell/small Thermowell adapter for 6.4 mm (1/4 in.) diameter 3.2 mm (1/8 in.) x SterilEnz pouch with
13
volume sample port 6.4 mm (1/4 in.) C-Flex tubing x 61 cm (24 in.) injection site assembly
Condenser bag
14–15 Condenser bag assembly (optional) –
assemblies (2)
Pressure sensor CPC AseptiQuick
16 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 8 cm (3 in.)
port aseptic connectors
Kleenpak Emflon II
12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 10 cm (4 in.) Reduced
17 Overlay gas sparger capsule and pressure
to 9.5 mm (3/8 in.) x 15.9 mm (5/8 in.) C-Flex tubing x 213 cm (84 in.)
transducer
12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 8 cm (3 in.) Reduced to
18 Inoculum addition 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 15 cm (6 in.) Reduced Plugged
to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 213 cm (84 in.)
12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 8 cm (3 in.) Reduced
19 Base addition to 6.4 mm (1/4 in.) x 11.1 mm (7/16 in.) C-Flex tubing x 213 cm (84 in.) Plugged
Reduced to 3.2 mm (1/8 in.) x 6.4 mm (1/4 in.) C-Flex tubing x 30 cm (12 in.)
19.1 mm (3/4 in.) x 25.4 mm (1 in.) C-Flex tubing x 213 cm (84 in.) splits
Media fill/auxiliary CPC AseptiQuik aseptic
20 to 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 61 cm (24 in.)
drain line connectors
and 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 61 cm (24 in.)
19.1 mm (3/4 in.) x 25.4 mm (1 in.) C-Flex tubing x 122 cm (48 in.)
splits to 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing x 61 cm
Bottom drain 1/4 in. MPC insert and
21 (24 in.) reduced to 6.4 mm (1/4 in.) x 9.5 mm (3/8 in.) C-Flex tubing
harvest Kleenpak (male)
x 30 cm (12 in.) and 12.7 mm (1/2 in.) x 19.1 mm (3/4 in.) C-Flex tubing
x 61 cm (24 in.)
22 Exhaust lines (2) Condenser bag assembly (optional) –
Condensate return
23 Condenser bag assembly (optional) –
line

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 167

Chapter 4 | System features and specifications

BPC packing information

Standard 5:1 S.U.B. BPC packaging is listed in Table 4.19, below.

Table 4.19. Standard 5:1 BPC packaging.

Outer packaging Supplied "flat-packed"; two polyethylene outer layers

Description, product code, lot number, expiry date on outer
Label
packaging, and shipping container
Sterilization Irradiation (25 to 40 kGy) inside outer packaging
Shipping container Durable cardboard carton
Documentation Certificate of Analysis provided with each lot for delivery

4.5 Additional system component part numbers

Tables 4.20–4.29 list part numbers for additional 5:1 S.U.B. system
components, such as drive shafts, load cell kits, and accessories.

Table 4.20. Drive shaft part numbers.

Description Cat. no.
50 L 1-piece aluminum drive shaft SV50959.10
100 L 1-piece aluminum drive shaft SV50959.18
250 L 1-piece stainless steel drive shaft SV50959.11
500 L 2-piece stainless steel drive shaft SV50959.12
1,000 L 3-piece stainless steel drive shaft SV50177.38
1,000 L 4-piece stainless steel drive shaft SV50177.39
2,000 L 2-piece carbon fiber drive shaft SV50959.21
2,000 L 4-piece carbon fiber drive shaft SV50959.20

Table 4.21. Cable management system part numbers.

Description Cat. no.
50 and 100 L S.U.B. cable management system SV50992.01
250 L S.U.B. cable management system SV50992.02
500 L S.U.B. cable management system SV50992.03
1,000 L S.U.B. cable management system SV50992.04

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 168

Chapter 4 | System features and specifications

Table 4.22. Load cell part numbers. 1,000 and 2,000 L S.U.B.s include load
cells as standard equipment. The following kits are for retro-fitting to 50, 100,
250, and 500 L S.U.B. systems.
Description Cat. no.
50–100 L S.U.B. load cell kit with summing block, no display SV50988.01
250 L S.U.B. load cell kit with summing block, no display SV50988.02
500 L S.U.B. load cell kit with summing block, no display SV50988.03

Table 4.23. Harsh mount load cell display part numbers for 50–500 L and
2,000 L systems.
Description Cat. no.
Mettler Toledo IND331 display, harsh mount style with analog
SV50177.306
interface (STD), 120 VAC US line cord/plug
Mettler Toledo IND331 display, harsh mount style with Allen-Bradley
SV50177.307
RIO interface, 120 VAC US line cord/plug
Mettler Toledo IND331 display, harsh mount style with Device Net
SV50177.308
interface, 120 VAC US line cord/plug
Mettler Toledo IND331 display, harsh mount style with Ethernet/IP
SV50177.309
and Modbus TCP interface, 120 VAC US line cord/plug
Mettler Toledo IND331 display, harsh mount style with Profibus
SV50177.310
interface, 120 VAC US line cord/plug

Table 4.24. Panel mount load cell display part numbers for 1,000 L
systems only.
Description Cat. no.
Mettler Toledo IND331 display, panel mount style with analog
SV50177.291
interface (STD), 120 VAC US line cord/plug
Mettler Toledo IND331 display, panel mount style with Allen-Bradley
SV50177.292
RIO interface, 120 VAC US line cord/plug
Mettler Toledo IND331 display, panel mount style with Device Net
SV50177.293
interface, 120 VAC US line cord/plug
Mettler Toledo IND331 display, panel mount style with Ethernet/IP
SV50177.294
and Modbus TCP interface, 120 VAC US line cord/plug
Mettler Toledo IND331 display, panel mount style with Profibus
SV50177.295
interface, 120 VAC US line cord/plug

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 169

Chapter 4 | System features and specifications

Table 4.25. Vent filter heater kit part numbers for use with Pall KA3 vent
filters. Includes vent filter heater, controller with water-tight closure, quick-
connects, and installation power cord.
Description Cat. no.
NEMA rated vent heater with programmable controller (100–120 VAC),
power cord. Includes low-temp. alarm, preset temp. 50°C, and power SV50191.11
cord with flying leads.
NEMA rated vent heater with programmable controller (200–240 VAC),
power cord. Includes low temp. alarm, preset temp. 50°C, and power SV50191.13
cord with flying leads.

Table 4.26. Vent filter heater kit part numbers for use with Meissner
Ultracap 10 in. vent filters. Includes vent filter heater, controller with water-
tight closure, quick-connects, and installation power cord.
Description Cat. no.
NEMA rated vent heater with programmable controller (100–120 VAC).
Includes low-temp. alarm, preset temp. 50°C, and 20 ft. NEMA 5-15 SV50191.16
power cord for US/Japan.
NEMA rated vent heater with programmable controller (200–240 VAC).
Includes low temp. alarm, preset temp. 50°C, and 20 ft. BS1363 power SV50191.17
cord for UK.
NEMA rated vent heater with programmable controller (200–240 VAC).
Includes low temp. alarm, preset temp. 50°C, and 20 ft. CEE7/7 power SV50191.18
cord for Europe.
NEMA rated vent heater with programmable controller (200–240 VAC).
Includes low temp. alarm, preset temp. 50°C, and 12 ft. IEC320 power SV50191.19
cord for 2,000 L S.U.B. control box.

Table 4.27. Condenser system part numbers (for 2,000 L systems only).
Description Cat. no.
Condenser system (120 V) including cart, chill plate and mounting post
SV50232.01
with filter brackets, TCU, and pump
Condenser system (240 V) including cart, chill plate and mounting post
SV50232.02
with filter brackets, TCU, and pump

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 170

Chapter 4 | System features and specifications

Table 4.28. AC and DC motor part numbers.

Description Cat. no.
50, 100, and 250 L S.U.B. DC motor SV50237.07
50, 100, and 250 L S.U.B. AC motor SV50237.16
500, 1,000, and 2,000 L S.U.B. DC motor SV50237.22
500 L S.U.B. AC motor SV50237.18
1,000 and 2,000 L S.U.B. AC motor SV50237.19

Table 4.29. Miscellaneous and accessory part numbers.

Component Cat. no.
Probe assembly with CPC AseptiQuik connector (non-sterile,
SH30720.02
for use in autoclave)
Probe assembly with Pall Kleenpak connector (non-sterile, for
SH30720.01
use in autoclave)
Heavy-duty tubing clamp SV20664.01
Stainless steel autoclave tray, for autoclaving probe
SV50177.01
assemblies
Probe clips SV50177P.01
Sterile sampling manifold with luer lock SH30845.01
Temperature/sample port SV20750.01
Sparge line support SV50177.19
Mobile stairs (for 2,000 L systems only) SV50935.01

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 171

 Maintenance and
5
troubleshooting

Chapter contents
5.1 Maintenance
5.2 Troubleshooting and frequently asked questions

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 172

Chapter 5 | Maintenance and troubleshooting

5.1 Maintenance
5.1.1 Routine maintenance

Environmental conditions, operating parameters, and adhering to

standard operating procedures as outlined in this user’s guide have
significant impact upon the useful life of your S.U.B. hardware system.
The following guidelines are based upon the standard operating
conditions outlined in this user's guide.

High-wear items such as bearings, seals, O-rings, and sterilization

valves common to conventional bioreactor systems have been
purposefully considered in the design of the construction of the S.U.B.
The S.U.B. system is inherently robust and requires low levels of routine
maintenance. Taking time between bioreactor runs to clean the exterior
of the hardware will improve the appearance and overall longevity of
the system. The drive motor is an industrial grade induction motor
with a permanently sealed and lubricated gear box. The drive shaft is
constructed to wear slightly with use and should be visually inspected
after each run. Visual inspection of wear components and following the
guidelines listed below will be sufficient to ensure dependable service.
Replacement parts are available.

5.1.2 Preventive maintenance

• Lightly coat the drive cap threads with food-grade anti-seize if the
motor cap becomes difficult to turn.
• For multiple-segment drive shafts without quick-connects, lightly
coat the threads with food-grade anti-seize during assembly.
• Replacement of the mixing motor is recommended every five years,
or as needed.
• Refer to the following section of this user's guide for expected drive
shaft longevity based on usage.

Replace worn drive shaft head assembly when the hex diameter at its
widest location measures equal to or less across the points (Figure 5.1).
Diameters are measured at the widest location across the points.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 173

Chapter 5 | Maintenance and troubleshooting

Hex diameter measurement

location

Figure 5.1. Shaft head assembly.

Under normal operating conditions, replace the drive shaft assembly

after 360 days of service, or refer to the drive shaft head wear
specifications in Table 5.1, below.

Table 5.1. Drive shaft head hex diameters.

S.U.B. system New hex diameter Minimum hex diameter

50, 100, and 250 L 14.9 mm (0.587 in.) ± 0.127 mm 14.4 mm (0.566 in.)

500 and 1,000 L 21.3 mm (0.839 in.) ± 0.127 mm 20.8 mm (0.820 in.)

Drive shaft longevity and replacement

For 50, 100, 250, 500, and 1,000 L S.U.B. systems, we recommend
replacing your drive shaft every 360 days of cumulative use.

2,000 L S.U.B. systems experience a greater amount of fatigue-related

stress than smaller systems due to the use of a longer drive shaft. If
you are operating a 2,000 L S.U.B. at a P/V ratio of 20 W/m 3, replace
your drive shaft every 180 days of cumulative use. If you are operating
3
a 2,000 L S.U.B. at 40 W/m , consultation with Thermo Scientific
engineers is required. For more information on the use of P/V ratios,
see section 3.6.5 of this publication. Note: Operating 2,000 L systems
below 50% working volume requires the use of safety interlocks and
speed governors. See the Warnings, safety, and warranty information
section in the front of this publication for more information.

Note: For warranty purposes, drive shaft use must be documented. A

drive shaft use log is provided in Appendix D of this publication.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 174

Chapter 5 | Maintenance and troubleshooting

5.2 Troubleshooting and frequently asked

questions

5.2.1 Hardware operation issues

Issue: The S.U.B. will not operate.

Solution: Check the power supply.

• Verify the position of the main electrical plug connection at the wall
outlet, the main power disconnect, and the emergency stop switch.
• Verify the condition of the main electrical circuit breaker at your
facility. If the protection breaker has been tripped, determine the
fault condition. The condition may exist where other electrical
systems are requiring current loads beyond those allowed by the
breaker. The S.U.B. system should be placed on its own electrical
circuit.
• Disconnect the main power cord. Inspect the electrical circuit
breakers and fuses inside the electrical box of the S.U.B. controller.
Determine the fault condition by visual inspection. If the fault
condition cannot be determined by visual inspection, contact the
manufacturer.

Issue: The S.U.B. temperature is below target or slow to respond.

Solution: Check the temperature controller and sensor.

• Verify that the temperature probe (RTD) is not loose, and has been
fully inserted into the BPC thermowell.
• Verify that the thermowell has been filled with sufficient glycerol to
aid in heat transfer.
• Verify that the temperature control unit (TCU) is operating, and all of
the ball valves are open.
• Verify that the system is filled with a sufficient volume of fluid. There
must be enough media (minimum volume) in the BPC to provide
contact with the container. Add more media if the BPC is not
touching the heater area.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 175

Chapter 5 | Maintenance and troubleshooting

Issue: Noise is being emitted from the mixer assembly.

Solution: No action is required.

The bearing port assembly supplied with the S.U.B. is an important
component in maintaining a sterile environment during cell growth.
The special seals used in the S.U.B. may generate some noise during
operation, particularly after the first day of operation. This noise may
vary in intensity and frequency, but generally has no significant effect
on performance or overall durability of the BPC during the intended life
of the product.

Issue: The mixer controller does not respond to user inputs.

Solution: Allow the speed to stabilize before using the keypad

interface.
• Adjusting the speed control too rapidly may require several seconds
for speed stabilization.
• Wait ten seconds, then attempt to adjust the speed at the keypad
interface.
• Verify the position of the input select switch of the variable
frequency drive (VFD). If the toggle switch is not in the middle
position, the controller will not be able to receive control inputs from
the control keypad on the front panel.

Issue: I typically use level sensors to control the volume and feed
rate or supplement during a bioreactor run; how would I do this
with the S.U.B.?

Solution: Use load cells or a scale to control volumes based upon weight.
The S.U.B. is not equipped with level sensors. However, the S.U.B. can
be set up to allow supplement feeds and volumes to be managed by
weight.

Issue: How do we ensure that we are using the correct components

and setting up the S.U.B. properly for 5:1 mixing?

Solution: Before you begin setup, verify that all hardware and components
are labeled for 5:1 mixing only (50–500 L systems only).
5:1 S.U.B.s require drive shafts and BPCs that are specifically made
for low-volume mixing. For 50–500 L systems, the motor mount, drive
shaft, and BPC should be engraved or labeled with "5:1 Mixing" and/or
a warning tag. If any of these components are not labeled for 5:1 use,
contact your sales representative immediately for a replacement.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 176

Chapter 5 | Maintenance and troubleshooting

5.2.2 Cell culture operation issues

Issue: Dissolved oxygen readings are low or slow to respond.

Solution: Check the physical condition of the dissolved oxygen (DO) probe,
the calibration of the probe, and gas flow rate into the S.U.B.
• DO probes require routine maintenance; replace the damaged
probe or membrane when necessary.
• Verify the DO probe calibration relative to setpoints of zero and
span.
• Inspect the line sets connected to direct spargers for restriction
(closed tubing clamp, pinched line, low supply pressure).

Issue: Dissolved oxygen readings are erratic or unstable.

Solution: Adjust the bioreactor controller to suit the volume of your S.U.B.
system.
• Many different parameters can affect the ability of a bioreactor
controller to maintain a target setpoint during process control.
Modern controllers utilize computer algorithms to adjust targeted
parameters; the most common technique is that of a tunable
controller that uses variables of the proportional integral derivative
(PID). Tuning these PID values to the specific characteristics of the
system dynamics will, in most cases, stabilize process parameters
to an acceptable level. We recommend that you consult the user
guide of the particular bioreactor controller you are using.
• A grounding reference to the media can be created by using a
grounding lead between the tank and the body of the stainless
steel DO probe or to the stainless steel connector (if present) on the
sample line of the BPC.

Issue: pH levels are questionable or out of range.

Solution: Verify the calibration of the probe and utilize either media or gas
buffers.
• pH levels can be managed in a similar manner to conventional
bioreactors once calibration of the probe is verified by use of an
off-line sample. Carbon dioxide gas sparged through the media or
headspace, biocarbonate levels in the media and the addition of
liquid titrant solutions all serve to manage the pH balance of the
bioreactor environment. See section 3.5.4 for more information on
probe calibration.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 177

Chapter 5 | Maintenance and troubleshooting

• A grounding reference to the media can be created by using a

grounding lead between the tank and the body of the stainless
steel DO probe or to the stainless steel connector (if present) on the
sample line of the BPC.

Issue: We are not achieving the cell growth we expected in the S.U.B.
while running under our normal bioreactor agitation and
sparging rates. What should we do?

Solution: Reduce agitation and sparging rates.

• Often low cell viability and cell growth can be attributed to
excessive sparging or agitation. We recommend that you reduce
the sparge rate compared to what you might use in a conventional
bioreactor. Gas flow rates supplied as overlay should also be
reduced as much as possible. Too much gas creates excess foam
and higher shear conditions. Provide only the level of agitation
needed (low viability and lysed cells), reduce agitation speed (cell
aggregation and settling), and increase agitation.
• Media formulation can also have a significant effect on cell culture
growth in the S.U.B. Surfactants, such as Pluronic, decrease shear
and increase kLa, but at a cost of increased foaming. Thermo
Scientific can offer custom media especially for the S.U.B. and your
specific cell line(s).

5.2.3 Sparging issues

Issue: There is excessive foam in the bioreactor headspace.

Solution: Alter the liquid surface tension related to the culture media and/or
sparge gas.
• A media supplement of antifoam can be used in the S.U.B. These
serve to lower the surface tension of the media and will reduce the
presence of foam.
• High sparge rates of air can result in the presence of excessive
foam. Testing has shown that sparging with oxygen will typically
result in a dramatic reduction of foam in the headspace.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 178

Chapter 5 | Maintenance and troubleshooting

Issue: The sparger does not seem to be working although gas is

present.

Solution: Allow the sparger membrane to purge.

• If the S.U.B. is filled with liquid and allowed to sit idle for extended
periods of time without gas being supplied to the sparger, liquid
can accumulate between the membrane and check valve. Various
media additives may restrict the membrane temporarily. Several
minutes of gas pressure being supplied to the sparger should
purge the membrane, allowing it to function properly.
• Certain operating conditions can create situations when the
sparger membrane may become restricted due to insufficient line
pressure from the bioreactor controller gas feed line. Increasing
the flow rate to one liter per minute, or momentarily raising the
pressure regulator outlet pressure to 0.34 bar (5 psi) may alleviate
the problem. Alternatively, several seconds at this higher pressure
will allow the membrane to purge pores that may be blocked due to
the presence of accumulated liquid.

5.2.4 Probe and connector issues

Issue: We forgot to introduce the pH and DO probes prior to media fill;

can we still make a sterile connection under these conditions?

Solution: Yes, as long as the clamps were closed on the aseptic connector
probe ports before liquid fill.
• The aseptic connectors must be dry to make the connection of the
probe assemblies. When media is already present in the S.U.B.,
follow the probe insertion procedures as outlined in section 3.5.3.
• Some fluid may enter the bellows when the probe is inserted into a
BPC already filled with media. This is normal and will not affect the
sterility of the system.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 179

Chapter 5 | Maintenance and troubleshooting

5.2.5 Other issues

Issue: The BPC seems overly tight.

Solution: Verify that the container is venting and inspect it for the cause of
overpressure.
• Reduce the inlet gas flow rate of overlay and direct sparger.
• Inspect the exhaust filter for restriction or blockage.
• Excessive foaming should be avoided for several reasons. If foam
levels are allowed to reach the exhaust filter, the filter will become
restricted, resulting in excessive internal pressure within the
confines of the S.U.B. This may cause product failure and bursting
of the BPC.

Issue: There is excessive pressure in the condenser bag for my

2,000 L system.

Solutions: Check for kinks in the exhaust tubing between the S.U.B. and the
condenser bag.
• Ensure that the condenser components are properly installed on
the condenser hardware.
• Inspect the tubing regularly for kinks, and monitor the pressure.

Check for liquid buildup in the condenser bag outlet tubing and/or
the vent filter.
• Ensure that the chiller power is on, the auto-restart option on the
chiller is activated, and the chiller setpoint and actual temperature is
correct.
• Ensure that the TCU coolant level in the chiller is at the maximum
capacity before use, as low levels of coolant will increase the chiller
plate temperature. Note: An infrared temperature reader can be
used to confirm that the chiller plates are near the TCU setpoint
(typically ± 2°C of the setpoint).
• Check that the batch flow rates do not exceed recommendations.
• Check coolant lines between the chiller and condenser plate for
abnormalities.
• Temporarily plug off vent filters (one at a time) while manipulating
the tubing to drain liquids back into the condenser bag.
• Continuously monitor the pressure in the S.U.B.

Check the exhaust tubing between the S.U.B. and the condenser
bag to ensure it is positioned correctly.
The braided exhaust hose tubing should not allow condensate to
collect, but should be able to drain into the vessel or condenser.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 180

Chapter 5 | Maintenance and troubleshooting

Check for liquid buildup in the condenser bag.

• Ensure that the pump power is on, the auto-restart option on the
pump is activated, and pump head is turning at the set speed, and
the pump is set to recommended speed (12–30 rpm).
• Ensure that the pump tubing is properly installed in the pump head,
and that there are no kinks or blockages in the liquid drain line
tubing.

Ensure that foam has not reached the condenser bag. If the foam
has reached the bag, reduce gas flow rates and add an anti-foam
agent. After the foam has been controlled, it will naturally dissipate and
drain out of the condenser bag.

Issue: There is excessive residue buildup in the condenser bag for my

2,000 L system.

Solution: Ensure that the TCU coolant level in the chiller is always at
maximum capacity before use. Low levels of coolant will increase
the chiller plate temperature, which results in excessive pressure and
residue buildup in the bag. Note: An infrared temperature reader can
be used to confirm that the chiller plates are near the TCU setpoint
(typically ± 2°C of the setpoint).

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 181

 General ordering
6
information
Chapter contents
6.1 Ordering instructions
6.2 Ordering/support contact information
6.3 Technical support information

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 182

Chapter 6 | General ordering information

6.1 Ordering instructions

BPCs and hardware components for the 5:1 Single-Use Bioreactor
(S.U.B.) can be ordered directly from Thermo Fisher Scientific. These
items include all components that have part numbers beginning with
the following digits:

• SH
• SV
• SUB

6.2 Ordering/support contact information

In the Americas and Asia
1726 Hyclone Drive
Logan, Utah 84321
United States
Tel: +1 435 792 8500
Email: customerservice.bioprocessing@thermofisher.com

In Europe
Unit 9 Atley Way
Cramlington, NE 23 1WA
Great Britain
Tel: +44 (1) 670 734 093
Fax: +44 (1) 670 732 537
Email: customerservice.bioprocessing@thermofisher.com

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 183

Chapter 6 | General ordering information

6.3 Technical support

Technical support for the 5:1 S.U.B. is available in a variety of formats.
Some or all of the following may be appropriate, depending on
individual experience and circumstances.

Technical service hotline and email

Contact your Thermo Scientific sales representative for general product
pricing, availability, delivery, order information, and product complaints.

Call +1 435 792 8500 (United States) or +44 (1) 670 734 093 (Europe,
U.K.) for direct and immediate response to overall product questions,
and product technical information (Technical Support). You can also
contact Tech Support by emailing:
techsupport.bioprocessing@thermofisher.com

Initial setup and operation

Appropriate technical support is available to assist in the initial setup
and operation of each S.U.B. system. If you require assistance in
setting up and operating your S.U.B. system, please inquire at the time
of purchase.

Training
Training can be provided for startup and operation of the S.U.B.
Contact your Thermo Scientific sales representative.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 184

Appendix A | Electrical receptacle installation

Appendix A—Installation of female electrical

receptacle for units with AC motors and
electrical control panels
1. In order to complete the installation for units with AC motors, the
facility must be equipped with an electrical housing of sufficient
size.
• Typically in the U.S. the plug will require a two-gang box when
using the adapter plate (supplied).
• For installations outside the U.S. (where an adapter plate is not
supplied), we recommend that an electrical control panel (E-Box)
be modified to accommodate the cutout dimensions as shown in
Figure A.1 below.

2. Verify that electrical power has been disconnected and locked out
for safety.

3. Verify that the holes for mounting the receptacle housing are
positioned properly. Center to center measurement of respective
mounting holes is 85 mm (3.35 in.) tall and 77 mm (3 in.) wide.

106 mm 77 mm
92 mm
38 mm 67 mm
62 mm

102.5 mm 81 mm 85 mm

5.5 mm DIA (4)

Figure A.1. Panel cutout.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 185

Appendix A | Electrical receptacle installation

4. Verify the condition of the three exposed wire leads and strip back
to expose new wire if needed.

5. Connect the wire leads on the receptacle (shown in Figure A.2

below) using the screw terminals, paying strict attention to obtain
the correct wiring position as it is labeled on the receptacle.
• Green (ground)
• White (common)
• Black in the U.S., Blue in the E.U. (hot)

Figure A.2. Female receptacle (blue for 240 VAC, yellow for 110 VAC).

6. If you are using the adapter mounting plate, secure it to the

selected facility electrical housing as per drawing (Figure A.3
below), otherwise proceed to Step 7.

7. Secure the electrical receptacle using the four supplied screw

fasteners.

8. Connect power back to the electrical circuit.

9. Test the circuit with multi-meter prior to making any connections to

the electrical receptacle.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 186

Appendix A | Electrical receptacle installation

3-4X #10X1 in. screws to mount plug to adapter

plate. Note: The plug will be offset to the left
or right if mounting to a two-gang box with
screws in opposite corners. If this is the case,
then one of the holes in the plug will line up
with the screw will need to be used in that hole.
Use other 2X #10X1 in. screws to plug holes
not used.

2X #8-32X 1 3/8 in. screws to mount to TYP

two-gang boxes. These boxes TYP have 2
screws in opposite corners.

4X #6-32X 1 1/4 in. screws to mount adapter

plate to TYP two-gang FS/FD box. Note: The
plug will be offset to the left or right using 4X
#10X1 in. screws. Use other 2X #10X1 in.
screws to plug holes not used.

Figure A.3. Adapter mounting plate.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 187

Appendix B | Mettler Toledo load cell calibration

Appendix B—Mettler Toledo IND331 display

load cell calibration instructions
Please refer to the instructions and reference material found in the
Mettler Toledo IND Terminal Technical Manual for specific procedures
and troubleshooting methods.

Verify the following before beginning load cell calibration:

• The Mettler Toledo IND display, load cell summing block, and load
cell transducers have been specified, installed, and configured
properly.
• The load cell transducers do not have the transport lockout nuts in
place (load cells must be ready for use prior to calibration).

The calibration accuracy achieved cannot exceed the precision of the

reference used for calibration.
• Field calibration is most often performed using calibrated reference
weights or flow meters for volumetric mass reference.
• Factory-trained technicians have the experience and tools
necessary to provide the best system performance and reliability. If
in doubt, contact your factory service representative.

Introduction
• Setup mode is accessed by pressing and holding the Print key for
approximately three seconds. See Chapter 2 of the Mettler Toledo
IND Terminal Technical Manual for further detail.
• Pressing Print is equivalent to pressing Enter. Use this key to
proceed through the sub-block numbers until you find your desired
choice.
• Press Select to toggle the values of the selected sub-block.
• The S.U.B. electrical schematic contains a table showing the sub-
blocks that have changed from the default settings.

Span calibration
The scale's span calibration can be determined with or without a
linearity adjustment. With linearity disabled, a single reference point
is used to calibrate the scale. This is the normal method of span
calibration. If linearity is enabled, an additional mid-range weight
reference point is added to the adjustment procedure. Linearity can be
enabled or disabled in the setup mode.

For further information, refer to the Mettler Toledo IND331 manual at:
http://mt.com

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 188

Appendix B | Mettler Toledo load cell calibration

Figure B.1. Span calibration.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 189

Appendix C | 2,000 L agitator operation and maintenance guidelines

Appendix C—2,000 L S.U.B. agitator

operation and maintenance guidelines
2,000 L Critical operating recommendations
WARNING: To prevent drive shaft breakage and maintain the
equipment warranty, follow the operating recommendations listed
below.
9 Verify that the motor variable frequency drive (VFD) is programmed
to accelerate and decelerate in a minimum of 60 seconds.
9 Use a controller agitation speed governor and safety interlocks to
prevent the system from running outside of recommended limits.
9 Verify that the drive shaft serial numbers match on all segments; do
not interchange shaft segments.
9 Always maintain a log history of drive shaft usage (see Appendix D)
and confirm that it has sufficient life remaining. If the age or
history of a shaft is questionable, it should be replaced. For more
information, see Maintenance in section 5:1 of this publication.
9 Only activate agitation after the BPC has been filled with media.
9 Use at least three operators to load the BPC into the 2,000 L S.U.B.
9 Fully inflate the BPC prior to insertion of the drive shaft. This will
keep the impeller tubing from stretching and being damaged.

Agitation speed recommendations

Table 3.3 in section 3.6.4 of this guide provides agitation speed
recommendations for all system sizes. 2,000 L bioreactor
recommendations are for systems with four-piece drive shafts, but two-
piece shafts will provide equivalent or greater longevity.

Note: The nominal agitation recommendations in Table 3.3 are based

on P/V values of 20 W/m 3. This is the suggested default parameter for
CHO cultivation. For more information on P/V calculation, see section
3.6.5 of this user's guide.

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 190

Appendix D | Drive shaft use log

Appendix D—Drive shaft use log

A sample log is provided below for tracking and documenting drive
shaft usage. Important note: For warranty purposes, users must
document proper drive shaft use.

Drive shaft serial number:

Vessel serial number:

Agitation setting Agitation setting Starting Finishing

Start date End date Number of days
(start) (stop) volume volume

Cumulative run time:

Thermo Scientific HyPerforma 5:1 Single-Use Bioreactor User's Guide | 191

Find out more at thermofisher.com/sub
For Research or Further Manufacturing. Not for diagnostic use or direct administration into humans or animals.
© 2021 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its
subsidiaries unless otherwise specified. Mettler Toledo is a trademark of Mettler Toledo AG. CPC and AseptiQuik are trademarks
of Colder Products Company. SmartSite is a trademark of Becton, Dickinson and Company. C-Flex is a trademark of Saint-Gobain
Performance Plastics. AC-Tech is a trademark of Lenze. Emflon and Kleenpak are trademarks of Pall Corporation. Meissner,
Ultracap, and Steridyne are trademarks of Meissner Filtration Products. SterilEnz is a trademark of PAW BioScience. Teflon is a
trademark of The Chemours Company. Velcro is a trademark of Velcro Company. Instron is a trademark of Illinois Tool Works, Inc.
Pluronic and Sigma-Aldrich are trademarks of MilliporeSigma. DOC0022 Rev. H 0221