Thermo Fisher Scientific HyPerforma 2:1 User Manual

HyPerforma 2:1 Single-Use
Bioreactor (S.U.B.) User’s Guide

DOC0014 • Revision H

January 2021

Contents

Warnings, safety, and warranty information 1
How to use this guide 7

Chapter 1 HyPerforma 2:1 Single‑Use Bioreactor overview 12

1.1 Introduction to the Single-Use Bioreactor 13

1.2 Hardware characteristics 15

1.2.1 S.U.B. hardware components 15

1.2.2 S.U.B. system features 17

1.2.3 Additional system components 18

1.3 End user and third-party supplied components 25

1.3.1 pH and DO probes 25

1.3.2 Controllers 26

1.4 BPC features 27

Chapter 2 Hardware assembly and setup 30

2.1 Initial installation preparation 31

2.1.1 Hardware shipment and setup 31

2.1.2 Hardware uncrating 31

2.1.3 Site preparation 31

2.2 Installation and setup 33

2.2.1 Preparing load cells 33

2.2.2 Leveling and connecting the system 35

2.2.3 Attaching the cable management system arm 37

Chapter 3 Operating information 38

3.1 General system operating information 39

3.1.1 BPC preparation 39

3.1.2 BPC handling instructions 39

3.1.3 BPC operating information 39

3.1.4 Hardware operating information 41

3.1.5 External data logging and control 44

3.2 BPC and drive shaft loading instructions for 50, 100,

and 250 L systems 44

3.2.1 Initial BPC loading steps for 50, 100, and 250 L

systems 44

Contents

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

systems 49

3.2.3 Final installation steps for 50, 100, and

250 L systems 53

3.3 BPC and drive shaft loading instructions for 500

and 1,000 L systems 55

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

systems 55

3.3.2 Drive shaft insertion for 500 and 1,000 L systems 61

3.3.3 Final installation steps for 500 and 1,000 L

systems 66

3.4 BPC and drive shaft loading, and condenser system

setup instructions for 2,000 L systems 69

3.4.1 Initial BPC loading steps for 2,000 L systems 69

3.4.2 Condenser system setup for 2,000 L systems 78

3.4.3 Drive shaft insertion for 2,000 L systems 89

3.4.4 Final installation steps for 2,000 L systems 95

3.5 Probe preparation and insertion 98

3.5.1 Preparation and sterilization 98

3.5.2 Making Kleenpak connections 99

3.5.3 Probe insertion 108

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 E-Boxes 112

3.6.5 Agitation rate calculations 113

3.6.6 Drive shaft rotation 117

3.6.7 Temperature control 118

3.6.8 Sparging strategy 119

3.6.9 pH probe calibration 123

3.6.10 DO probe calibration 123

3.6.11 Checkpoints prior to inoculation 124

3.6.12 Cell inoculation 124

3.6.13 Volume scale up 124

3.6.14 In-process checkpoints 125

Contents

3.6.15 BPC sampling 125

3.6.16 Dispense and harvest 128

3.6.17 BPC disposal 128

3.6.18 S.U.B. shutdown 129

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 130

3.7.3 Pressure monitor verification (when present) 130

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 500–1,000 L systems 133

4.1.3 Design features for 2,000 L systems 134

4.2 Hardware specifications 135

4.3 E-Box features 153

4.4 BPC specifications 154

4.5 Additional system component part numbers 175

Chapter 5 Maintenance and troubleshooting 179

5.1 Maintenance 180

5.1.1 Routine maintenance 180

5.1.2 Preventive maintenance 180

5.2 Troubleshooting and frequently asked questions 182

5.2.1 Hardware operation issues 182

5.2.2 Cell culture operation issues 185

5.2.3 Sparging issues 186

5.2.4 Probe and connector issues 187

5.2.5 Other issues 188

Chapter 6 General ordering information 190

6.1 Ordering instructions 191

6.2 Ordering/support contact information 191

6.3 Technical support 192

Contents

Appendices Appendix A—Installation of female electrical receptacle

for units with AC motors and electrical boxes 193

Appendix B—Mettler Toledo IND331 display load cell

calibration instructions 196

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

maintenance guidelines 198

Appendix D—Drive shaft use log 199

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
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 the user’s guide before operating; failure to do so
could result in injury and potential loss of product.

™

HyPerforma™ Single-Use Bioreactor (S.U.B.)

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 Thermo Fisher Scientific service
personnel only. Thermo Fisher Scientific recommends using standard

lockout procedures when working on electrical components. The main
breaker on the electrical 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 non-conductive materials,
such as non-conductive gloves, is recommended.

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

Warnings, safety, and warranty information

WARNING: Rotating parts—entanglement hazard.

Rotating and moving parts 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

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

Warnings, safety, and warranty information

WARNING: Hot surface—do not touch.

The heating jacket is designed to heat the inner vessel wall. Normal
operating conditions 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 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, non­condensing

• Storage: –25°C to 65°C

• Installation category II (over voltage) in accordance with IEC 664

• Altitude Limit: 2,000 meters

HyPerforma 2:1 Single‑Use Bioreactor User's Guide | 3Thermo Scientific

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. is NOT plugged into a GFCI outlet.

Water jacket vessel information

S.U.B. hardware unit with water jacket has 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

HyPerforma 2:1 Single‑Use Bioreactor User's Guide | 4Thermo Scientific

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.

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

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.

HyPerforma 2:1 Single‑Use Bioreactor User's Guide | 6Thermo Scientific

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 2:1 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

1.4 05/2016 -- Initial Release S. Jelus

B 12/2016

B 12/2016 4.2

C 12/2016

C 12/2016 2.2

C 12/2016 3.6.4

C 12/2016 5.1.2

C 12/2016 3.4

C 12/2016 3.4

C 12/2016 3.4

C 12/2016 3.4

C 12/2016 4.2

C 12/2016 4.2

C 12/2016 4.5 Added drive shafts as accessories S. Jelus/E. Hale

C 12/2016 Appendix D

How to Use This

Guide

Warnings and

Safety

Added How to Use This Guide section E. Hale

Fixed Electrical Power Supply Requirement in
Specifications section

Added information about safety interlocks to Warnings
and Safety section

Added serial number information and photo of ends of
multiple-section drive shafts

Added warning note about agitation rate and volume
requirements, and the use of safety interlocks

Added measurement to Table 1.10 for 2,000 L drive
shafts and cross-reference to Appendix D

Added information about 2-piece drive shaft and a note
about position of impeller tubing inside the BPC

Added serial number information and photo of ends of
multiple-section drive shafts

Added a note about not pushing drive shaft straight into
the assembly when loading

Added information and Figure 2:105 to illustrate proper
insertion of drive shaft

Added information about 2-piece drive shaft to 2,000 L
specifications

Added ceiling height requirements for 2-piece drive
shaft and detail about mixing speed to 2,000 L
specifications

Added Appendix D—2,000 L S.U.B. Agitator Operation
and Maintenance Guidelines

E. Hale

S. Jelus

S. Jelus/E. Hale

S. Jelus/E. Hale

S. Jelus

S. Jelus/E. Hale

S. Jelus/E. Hale

S. Jelus/E. Hale

S. Jelus/E. Hale

S. Jelus/E. Hale

S. Jelus/E. Hale

S. Jelus/E. Hale

D 02/2017 Appendix D Removed Table D.1 in Appendix D E. Hale

D 02/2017 3.6.5

Moved sections from Appendix D to new
Agitation Rate Calculations section

HyPerforma 2:1 Single‑Use Bioreactor User's Guide | 7Thermo Scientific

E. Hale

How to use this guide

Document change information (continued)

Revision Date Section Change made Author

D 02/2017 3.3

D 02/2017 3.4

D 02/2017 3.4

Changed “500–1,000L Electric Resistive Heater” to “500–1,000 L
Volumes” in BPC Loading section

Changed “BPC Loading 2,000L Water Jacket” to “BPC loading 2,000 L
volume” in BPC loading section

Moved “Securing access doors” step after port alignment step in 2,000
L BPC loading

E. Hale

E. Hale

E. Hale

D 02/2017 Appendix E Moved drive shaft log from Appendix D to Appendix E E. Hale

D 02/2017 3.3

Moved “Securing access doors” step from 50–250 L BPC loading to
500–1,000 L BPC loading

E. Hale

D 02/2017 6.2 Updated addresses, phone numbers, and email address E. Hale

D 04/2017

Warnings and

safety

D 04/2017 4.2

D 04/2017 4.2

Updated “Use of Agitation speed governors and safety interlocks” in
Warnings and safety

Changed “Maximum mixing rate” to “Agitation speed range” in hard­ware specifications

Added “Minimum acceleration and deceleration rate” to 2,000 L hard­ware specifications

E. Hale

E. Hale

E. Hale

D 04/2017 4.5 Corrected part numbers for 2,000 L S.U.B. drive shafts E. Hale

D 04/2017 3.6.3 Updated media fill instructions E. Hale

D 04/2017 5.1.2

D 05/2017

Warnings and

safety

D 05/2017 3.2, 3.3, 3.4

Updated drive shaft replacement intervals for 20 and 40 W/m
"Drive shaft longevity and replacement"

Added potentially explosive atmosphere (ATEX) warning E. Hale

Added step about removing plastic insert in the thermowell before
inserting RTD

3

P/V in

E. Hale

E. Hale

D 05/2017 2.1, 3.1 Updated 2,000 L load cells and unlocking instructions E. Hale

D 05/2017 Chapter 1, 4.2

Removed electric resistive heater options for 500, 1,000, and 2,000 L
systems

E. Hale

D 05/2017 4.4 Corrected format for units of measurement in BPC specifications E. Hale

E 06/2018

Warnings and

safety

Replaced the images for the following warning labels: "Read and under­stand the user's guide..." and "Entanglement hazard"

K. Leeman

Warnings, safety,

E 06/2018

and warranty

Added warranty information and use restrictions K. Leeman

information

E 06/2018 3.6.5

E 06/2018 3.6.5 Changed footnote in Table 1.7 from "40 W/m

Revised Graph 1.2 by changing 2,000 L line to 750 L, and 1,000 L line
to 375 L

3

" to "> 20 W/m3" K. Leeman

K. Leeman

Added information about protection against drive shaft instability,

E 06/2018 3.6.5

including a graph depicting regions of potential agitator harmonics and

K. Leeman

cavitation for liquid working volumes of the 2,000 L S.U.B.

E 06/2018 3.6.5 Added 20–40% fill agitation rates for all S.U.B. sizes to Table 1.8 K. Leeman

E 06/2018 3.6.5 Changed first footnote in Table 1.9 from "40 W/m

3

" to "> 20 W/m3" K. Leeman

E 06/2018 5.1.2 Removed 2,000 L row from Table 1.10 K. Leeman

HyPerforma 2:1 Single‑Use Bioreactor User's Guide | 8Thermo Scientific

How to use this guide

Document change information (continued)

Revision Date Section Change made Author

Under "Drive Shaft Longevity and Replacement," added "of cumulative
use" after "we recommend replacing your drive shaft every 360 days."

E 06/2018 5.1.2

E 06/2018 3.4

E 06/2018 3.1 Revised Table 1.3 to reflect the recommended heating times for S.U.B.s K. Leeman

E 06/2018 3.6.4

E 06/2018 5.1.2

E 06/2018 4.2

E 06/2018 -- Reformatted using new template and reorganized chapters/content E. Hale

E 06/2018 4.2

E 06/2018 5.2 Added FAQ about excessive residue buildup in condenser bag E. Hale

In the second sentence of the second paragraph, verbiage was changed
to "...every 180 days of cumulative use." In the first sentence of the
note, added "at < 50% working volume"

Replaced Figure 2.105 with an image to reflect the deep pocket
impeller change

Added note to Tables 1.8 and 1.9 about system recommended speed/
volume control parameters

Added Table 1.11 and related note describing 2-piece drive shaft
operating parameters for 2,000 L S.U.B.s

Updated "Operating temperature" in specifications for all sizes to
"Ambient to 40 ± 0.5°C (104 ± 0.9°F)"

Corrected ceiling height requirement for 2,000 L S.U.B. 4-piece drive
shaft loading, and added noise level to specifications for all S.U.B. sizes

K. Leeman

K. Leeman

K. Leeman

K. Leeman

E. Hale

E. Hale

E 06/2018

E 06/2018 1.2.3, 3.4.2 Added side-mounted condenser system illustration and information E. Hale

E 06/2018 3.2.1

E 06/2018 -- Removed references to 4-piece drive shafts for 2,000 L S.U.B.s E. Hale

E 08/2018

F 11/2018

F 11/2018 -- Removed references to metal probe clips E. Hale

F 11/2018 2.1.3, 3.6.4, 4.3 Updated text about and images of the E-Box E. Hale

F 11/2018

F 11/2018 Appendices

F 11/2018 2.2.3, Various

F 12/2018 3.1.4, 3.6.4 Edited sentence (3.1.4) and reworded step #2 (3.6.4) E. Hale

F 12/2018 4.2 Added tolerance to "Agitation speed range" in all specifications E. Hale

F 12/2018 3.7.1

How to use this

guide

Warnings, safety,

and warranty

information

Warnings, safety,

and warranty

information

How to use this

guide

Added "Abbreviations/acronyms" section E. Hale

Updated image of media ground clip connection for 50–250 L BPC
loading

Added seismic guidance K. Leeman

Added emphasis to "Electrical connections" section, changed "certified
personnel" to "Thermo Fisher Scientific service personnel," and updated
ATEX warning

Changed "Input into Thermo Scientific publications" section to
"Questions about this publication"

Removed Appendix B (AC-Tech variable speed drive settings) and
renamed Appendices C, D, and E to Appendices B, C, and D

Removed section 2.2.3 (Attaching the cable management system
arm) and edited images showing the arm

Updated expected accuracy in "Mixing speed verification" to ± 1.5
rpm or 1% of setpoint, whichever is greater

E. Hale

E. Hale

E. Hale

E. Hale

E. Hale

E. Hale

HyPerforma 2:1 Single‑Use Bioreactor User's Guide | 9Thermo Scientific

How to use this guide

Document change information (continued)

Revision Date Section Change made Author

G 10/2019 4.2, Various Minor revisions and updated cart length demention on Figure 4.10 T. Golightly

G 06/2020 -- Minor formatting revisions T. Golightly

Warnings, safety,

G 06/2020

G 06/2020 3.2.1 Removed former Step 13, and removed former Figure 3.11 T. Golightly

G 06/2020 3.2.1

G 06/2020 3.3.1, 3.4.1 Added a CAUTION note to the BPC loading instructions T. Golightly

H 11/2020 4.2 Corrected the overall width, length, and height in Tables 4.2 and 4.4 E. Hale

H 11/2020 4.2 Replaced Figures 4.7–4.10 with updated dimensions T. Golightly

H 11/2020 1.3.1, 4.4

H 11/2020 4.4

and warranty

information

Added Warnings for Pinch Hazard and Tipping Hazard T.Golightly

Added a CAUTION note below Step 12 for the BPC loading
instructions

Updated "Finesse" and "PreSens" sensors to "Hamilton" sensors in
Tables 1.1 and 4.23

Removed the "PreSens and Finesse" sensors and replaced with
"Hamilton" sensors in Table 4.23

T.Golightly

T. Golightly

T. Golightly

HyPerforma 2:1 Single‑Use Bioreactor User's Guide | 10Thermo Scientific

How to use this guide

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 2:1 S.U.B. Validation
Guide (DOC0016)

Thermo Scientific HyPerforma 2:1 S.U.B. Data Sheets
(for various sizes)

Information about validation
procedures

Product descriptions and ordering
information

Abbreviations/acronyms

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

BPC BioProcess Container
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

HyPerforma 2:1 Single‑Use Bioreactor User's Guide | 11Thermo Scientific

Chapter 1 | S.U.B. overview

1

HyPerforma 2:1
Single‑Use Bioreactor
overview

Chapter contents

1.1 Introduction to the Single‑Use Bioreactor

1.2 Hardware characteristics

1.3 End user and third‑party supplied components

1.4 BPC characteristics

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 12Thermo Scientific

Chapter 1 | S.U.B. overview

1.1 Introduction to the Single‑Use Bioreactor

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.

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, or
resistive heater for 50, 100, and 250 L systems

2. S.U.B. BioProcess Container (BPC), which 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, drive shaft, and impeller

Figure 1.1. 50–500 L S.U.B.s.

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 13Thermo Scientific

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 or resistive tank
on casters (2,000 L S.U.B.s are not on casters). Water jacketed heating
is an option for all tank sizes, and resistive heating is available for 50,
100, and 250 L tanks. 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 on 1,000 and 2,000 L systems,
and are optional for smaller 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 cGMP and is
supported by qualification and validation information. No reuse cleaning
is required. Innovative, proprietary technology allows for the integration
of the mixing shaft and pH and 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 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 selected and supplied by
the end user or by Thermo Fisher Scientific.

This user’s guide covers the setup, operation, maintenance, and
troubleshooting of all 2:1 S.U.B. systems in the following volumes—50,
100, 250, 500, 1,000, and 2,000 L.

Note: This guide is for S.U.B. systems that operate at a minimum
working volume of 50% (also known as 2:1 mixing). If you are using
a S.U.B. system capable of operating at 20% working volume (5:1
mixing), refer to the HyPerforma 5:1 Single‑Use Bioreactor User's Guide
(DOC0022).

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 14Thermo Scientific

Chapter 1 | S.U.B. overview

1.2 Hardware characteristics

1. 2.1 S.U.B. hardware components

Figures 1.2 and 1.3 below illustrate all available components of a water‑
jacketed 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.

1

2

12

4

13

3

6

8

9

10

Figure 1.2. Front/side view of 500 L S.U.B. Figure 1.3. Back view of 500 L S.U.B.

1. Exhaust vent filter holder

2. Mixing assembly with shield

3. Mixer motor

4. Bearing port receiver with clamp

5. Liquid sight windows

6. Drive shaft, stored

7. Electrical control panel (E-Box), optional

8. Probe hanger bracket

9. Probe access windows

10. Leveling casters

5

7

15

11

17

11. Cart assembly

12. 0.95 cm (3/8 in.) Dimpled water jacket (not present
in resistive 50, 100, and 250 L S.U.B.s)

13. 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

14. Stainless steel outer support container

15. Bleed valve

16. Bottom cutouts/pins for BPC attachment/alignment

17. Quick connect water inlet/outlet ports (for water­jacketed S.U.B.s only)

14

16

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 15Thermo Scientific

Chapter 1 | S.U.B. overview

2

Figures 1.4 and 1.5 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. See section 4.1.3 for a complete illustration of a
1,000 L S.U.B.

1

12

3

4

5

6

7

11

8

9

10

14

15

16

13

17

Figure 1.4. Front/side view of 2,000 L S.U.B. Figure 1.5. Side/bottom view of 2,000 L S.U.B.

1. Bag lift assembly

2. Auxiliary emergency stop (E-Stop)

3. Mixing assembly with shield

4. Mixer motor

5. Standard tool set

6. Load cell display

7. Electrical control panel (E-Box)

8. Probe access window

9. Probe clips

10. Bottom cutouts/pins for BPC alignment

11. Pneumatic bag lift control

12. Water jacket

13. Stainless steel outer support container

14. Load cell summing block

15. Quick connect water inlet/outlet ports

16. Load cells (3)

17. Sparge plate access

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 16Thermo Scientific

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
keypad interface on the control panel. 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. For 50, 100, and 250 L systems with resistive heaters and
Thermo Scientific E‑Boxes, temperature setpoints can be adjusted
via the temperature controller located on the front panel of the S.U.B.
E‑Box. This controller is pre‑programmed to avoid overshoot during
heat‑up, and to maintain a target temperature of ± 0.5°C based on
the set value display. The process temperature is measured by means
of a supplied resistive temperature detector (RTD) (pt‑100) that is
inserted into the thermowell of the S.U.B. BPC. Water jacket system
temperature control is maintained through the temperature control unit
(TCU).

Heating performance

Heating times for the S.U.B. systems vary based on 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 and heaters in 50–250 L resistive systems 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.

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 17Thermo Scientific

Chapter 1 | S.U.B. overview

1.2.3 Additional system components

Drive shafts

The S.U.B. drive shaft is detachable and reusable. It is inserted into
the BPC through the hollow pass‑through of the motor assembly, into
the bearing port, through the tubing sleeve inside the BPC, and into
the polyethylene impeller. Drive shaft rods may be made of aluminum,
stainless steel, or carbon fiber, depending on the size of the vessel and
the strength requirements.

As a general rule, drive shafts should be replaced after 360 days of
service, or as specified in Chapter 5 of this publication. Always keep a
log of actual drive shaft usage. Appendix D includes a form that can be
used for this purpose.

AC and DC motors

AC and DC motor options are available to help tailor the system to
specific needs. The DC motor operates at a lower voltage and, when
integrated with a controller system that receives sensor feedback,
provides 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, and must be
specified by the end user.

The AC motor may be used with the Thermo Scientific E‑Box, includes
the variable frequency drive, and is controlled using either the provided
keypad or a controller specified by the end‑user.

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 1.6), 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.

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 18Thermo Scientific

Chapter 1 | S.U.B. overview

Figure 1.6. Vent filter heater.

Condenser systems (for 2,000 L units only)

The condenser system supports the effective use of 2,000 L S.U.B.s,
and condenser systems with a cart assembly are also available as
an auxiliary component for other S.U.B. sizes. The condenser system
efficiently condenses exhaust gases and transfers condensate back
into the bioreactor, preventing potential vent filter blockage and
reducing fluid loss due to evaporation. It is offered in both single
and double chill‑plate formats. 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.

The condenser system protects against filter blockage by condensing
out moisture prior to exhaust gases reaching the vent filters. BPCs are
not intended to operate under pressure, and fouled (blocked) exhaust
filters lead to bag pressurization. While vent filter heaters may prevent
condensate buildup in many instances, in larger bioreactors (such
as the 2,000 L S.U.B.) this becomes less effective. Condensing out
the moisture first is a more reliable method for preventing liquid from
reaching the filters.

The S.U.B. condenser system with cart assembly Figure 1.7) consists
of the following components:

• Cart and brackets provide convenient means of organizing and

transporting key working elements of the condenser system.

• Chill plates secure disposable double chamber condenser bags

to cool exhaust gases. Up to two plates can be used per system.

• Peristaltic pump, for returning condensate to the bioreactor.

• Temperature control unit (TCU, also referred to as a chiller),

which circulates water to cool the condenser plate.

• Condenser disposables include the BPC (double‑chambered

bag), tubing, and exhaust filters though which the exhaust gases
flow and are chilled, and in which the condensate collects and is
returned to the bioreactor.

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 19Thermo Scientific

Chapter 1 | S.U.B. overview

Condenser disposables Condenser hardware

Complete condenser
system

Exhaust
vent
filters

Condenser
bag gas
outlet port

Dual chamber
condenser bag

Condenser bag
liquid drain ports

Exhaust line
from S.U.B.

Condenser
return line
back to S.U.B.

Condenser
bag gas
inlet port

Gripping
tabs

Alignment
holes

Figure 1.7. Overview of condenser system cart assembly option for 2,000 L S.U.B.s.

Filter straps

Condenser
plate assembly

Dual
headed
peristaltic
pump

Closed bath
recirculating
chiller

Filter bracket
assembly

Condenser post
assembly

Post receivers

Cart
assembly

Side‑mounted condenser systems (Figure 1.8) are only available for
2,000 L S.U.B.s, and attach directly to the outer support container.

Figure 1.8. Side-mounted condenser
system option for 2,000 L S.U.B.s.

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 20Thermo Scientific

Chapter 1 | S.U.B. overview

Load cells

Load cells, which are used to determine the weight of the contents
of a S.U.B., are installed on standard 1,000 and 2,000 L S.U.B.
systems, and are available as an option for 50–500 L units. Load cell
retro‑fit kits can also be added to existing S.U.B. units by a certified
service technician. Note: 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 1.9).

Figure 1.9. Load cell system overview.

Load cells are typically radial‑mounted in sets of three. The mounting
location (Figure 1.10) varies slightly for each size in order to allow easy
access to the bottom drain or sparging mechanisms and tubing.

Load cell

Figure 1.10. Load cell location.

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 21Thermo Scientific

Chapter 1 | S.U.B. overview

Probe integration

The autoclave tray (Figure 1.11) 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

Note: Figure 1.11 shows the autoclave tray used for probes with

™

Kleenpak™ aseptic connectors. Your system may use CPC™

Pall
AseptiQuik

™

aseptic connectors instead. Consult your sales

representative for more information on AseptiQuik connectors.

Handle

Probe assembly

Autoclave tray
for probe kits

Figure 1.11. Autoclave tray and probe assembly.

The probe assembly (Figure 1.12) 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 a Kleenpak
aseptic connector, molded bellows cover, and threaded probe adapter.

Aseptic
connector

Molded bellows cover

Threaded probe
adapter

Figure 1.12. Probe assembly.

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 22Thermo Scientific

Chapter 1 | S.U.B. overview

Channels for
sparge lines

Cable management systems

The cable management system (Figure 1.13) is available as on option
on 50, 100, 250, 500, and 1,000 L units. It is used to organize various
lines and includes the following components.

• Internal channel for sparge lines

• External channels for feed and base addition lines

• Harvest line hook

Channels for
feed and base
addition lines

Harvest line
hook

Figure 1.13. 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 resistance temperature

detectors (RTD) calibration/validation

• Sparge line support—Keeps the drilled hole sparge line in
a vertical position for optimal gas flow (Figure 1.14). For more
information see section 2.2, Installation and setup.

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 23Thermo Scientific

Chapter 1 | S.U.B. overview

Figure 1.14. Sparge line support.

• Heavy-duty tubing clamps (typically four or five)—Tu bi ng
clamps (Figure 1.15) 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.

Figure 1.15. 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.

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 24Thermo Scientific

Chapter 1 | S.U.B. overview

Table 1.1. Manufacturers and models of compatible pH/DO probes.

Probe lengths (from O-ring to tip) must not exceed 235 mm O-ring to probe tip

Probe manufacturer

and type

AppliSens DO Z010023525

AppliSens pH Z00102 3551

Mettler Toledo DO

Mettler Toledo pH

Broadley-James DO D140-B220-PT-D9

Broadley-James pH F-635-B225-DH

Hamilton DO 237542

Hamilton pH 238633-2543

Note: Consult the probe manufacturer’s website for appropriate probe cable connection and part number.

1.3 End user and third‑party supplied

components

1. 3 .1 pH and DO probes

Table 1.1 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.

Part number Diameter Thread type

12 mm

(0.47 in.)

12 mm

(0.47 in.)

InPRO 6800/12/220, PN

52200966

405-DPAS-SC-K8S/225, PN

104054481IG

12 mm

(0.47 in.)

12 mm

(0.47 in.)

12 mm

(0.47 in.)

12 mm

(0.47 in.)

12 mm

(0.47 in.)

12 mm

(0.47 in.)

13.5 PG

13.5 PG

13.5 PG

13.5 PG

13.5 PG

13.5 PG

13.5 PG

13.5 PG

Print/lit.

length

235 mm

(9.25 in.)

235 mm

(9.25 in.)

215 mm

(8.46 in.)

195 mm

( 7.6 7 in . )

215 mm

(8.46 in.)

225 mm

(8.85 in.)

225 mm

(8.85 in.)

225 mm

(8.85 in.)

Actual
length

235 mm

(9.25 in.)

235 mm

(9.25 in.)

215 mm

(8.46 in.)

219 mm

(8.62 in.)

214 mm

(8.42 in.)

219 mm

(8.62 in.)

220 mm

(8.66 in.)

220 mm

(8.66 in.)

HyPerforma 2:1 Single-Use Bioreactor User's Guide | 25Thermo Scientific