Thermo Fisher Scientific HyPerforma 2:1 User Manual

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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

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

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 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

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

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

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

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

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

P/V in

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

" 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

" 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

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

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

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

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.

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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).

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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.

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

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)

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Chapter 1 | S.U.B. overview

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.

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

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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.

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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.

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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.

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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.

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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.

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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.

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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.

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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.

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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

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.)

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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 Equipment Turnover Package (ETP) supplied with the HyPerforma S.U.B. Companies that offer control solutions in either cGMP 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 2:1 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.

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Chapter 1 | S.U.B. overview

1.4 BPC features

The cell culture itself is contained inside the BPC (Figures 1.16–1.18). 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. The bioreactor BPC is supplied gamma irradiated.

Operating pressure

The S.U.B. BPC does not operate as a closed system, as it has both inlet and exhaust ﬁlters that are utilized to maintain an environment for cells to grow without concern for contamination. However, conditions can be encountered when gas inlet ﬂow rate may exceed exhaust ﬂow rate. This may be encountered in the unlikely event of a pressure regulator failure on a gas feed, or when excessive foaming creates conditions of vent blockage. The S.U.B. BPC is not rated as a pressure vessel [gas pressure should not exceed 0.03 bar (0.5 psi) within the BPC]. Custom BPCs can be ordered with an optional single‑use pressure transducer for monitoring the pressure within the S.U.B. (supplied standard with 1,000 and 2,000 L systems).

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.

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 using a tubing welder, the quick connect, or ﬁtting provided. Manipulation of the BPC as the last few liters of media are removed can minimize liquid hold‑up in the S.U.B.

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Chapter 1 | S.U.B. overview

Sparging

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 is a method of providing for the oxygen requirements of eukaryotic cell cultures. The standard S.U.B. BPC incorporates a unique single‑use dual sparging design that allows for optimal aeration of the culture process and effective carbon dioxide stripping.

Connections

Multiple aseptic connection options exist for S.U.B. users. Standard BPCs include tubing welder sections, quick connects, and Pall Kleenpak connections. Note: CPC AseptiQuik connectors are also available. 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.

Figures 1.16–1.18 on the following page, show the features of all sizes of 2:1 S.U.B. BPCs. For more information about the components labeled in the ﬁgures, see Table 1.2.

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Chapter 1 | S.U.B. overview

7-8

Figure 1.16. Standard BPC for 50, 100, and 250 L systems.

Figure 1.17. Standard BPC for 500 and 1,000 L systems.

Figure 1.18. Standard BPC for 2,000 L systems.

Table 1.2. BPC information for Figures 1.16–1.18.

Item Component Description

1 Exhaust vent ﬁlter Single-use capsule ﬁlter for exhaust gas exchange

2 Gas overlay port Protected by gas ﬁlter

3 Ports For addition of media and other liquids

4 Seal/bearing assembly Links with motor mixer and allows impeller to turn while retaining integrity of the BPC

5 Impeller

Injection-molded plastic, linked to seal/bearing assembly by C-Flex tubing contact material of the shaft

6 Ports with Kleenpak connectors For integration of standard 12 mm (0.47 in.) monitoring pH and DO probes

7-8

7 Temperature RTD port For integration of temperature probe while retaining integrity of the BPC

8 Sampling port For needleless sampling or connection to sampling manifold

9 Drain port For draining the S.U.B.

10 Gas sparge lines

Sparger integrated into the chamber and protected by gas ﬁlters; dual micro sparger (porous frit) and macro sparger (with open pipe or drilled hole) options are available

Integrates with chiller plate to remove condensate from exhaust; supports effective

11 Condenser system bag

use of 2,000 L systems (can also be used with smaller sizes) S.U.B.s that require custom gassing strategies and demand higher exhaust rates and longer durations

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Chapter 2 | Hardware assembly and setup

Hardware assembly and setup

Chapter contents

2.1 Initial installation preparation

2.2 Installation and setup

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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 unpacking and assembly instructions, as well as 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 electrical control panel

• Drive shaft, resistance temperature detector (RTD), four probe

• Equipment Turnover Package (ETP), provided on a USB drive

(E‑Box)]

brackets, and standard tool set (spanner wrench and torque wrench)

(shipped separately)

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 E-Boxes

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.

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Chapter 2 | Hardware assembly and setup

Electrical connections for systems with DC motors

S.U.B. units using DC motors are not supplied with 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, refer to the ETP, which is provided on a USB drive.

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

VFD breaker

Main power breaker

Temp. display breaker

Pressure breaker

E-Stop power breaker

Continuous power breaker

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Chapter 2 | Hardware assembly and setup

2. Verify that the three‑way motor controller switch is in the middle

3. Close the E‑Box. Use a screwdriver to lock the E‑Box before

4. For S.U.B. hardware units purchased with factory‑installed load

position. For reference:

• The middle position enables the speed control keypad

• The top position is for 0–10 V controllers

• The bottom position is for 4–20 mA controllers

Verify that the position of the two‑way temperature control switch is in the up position. This will enable the PID temperature controller.

continuing.

cells, the load cells are shipped in the locked position (threaded up) for equipment protection. Refer to the load cell preparation instructions later in this section for more information.

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.

Use the following 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.

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.

2. To unlock the load cells, remove and discard the delrin slip ring (if

present). Remove the tri‑clamp. Use the small end of the supplied tool (Figure 2.3) to loosen the lockout nut 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.

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Chapter 2 | Hardware assembly and setup

Lockout nut

38.1 mm (1. 5 in.) tri-clamp

Lockout post

Drive shaft cap end

Delrin slip ring

Figure 2.2. Close-up view of load cells.

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.

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Chapter 2 | Hardware assembly and setup

2.2.2 Leveling and connecting the system

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

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

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

power requirements of the S.U.B. and ancillary components, such as controllers or pumps.

run.

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. 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 resistive 50, 100, and 250 L systems, verify that the

temperature controller is off. The display should be ﬂashing in the stand‑by position.

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Chapter 2 | Hardware assembly and setup

10. 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.

Inlet port

11. 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.

Outlet port

Figure 2.5. Inlet and outlet ports.

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Chapter 2 | Hardware assembly and setup

12. 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 component holds the sparge line vertically for maximum effectiveness. The sparge line can be wound through the coil of the holder to keep the sparger properly oriented.

Figure 2.6. Sparge line support.

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

• Special quick connect designs to reduce joint fatigue in multiple‑

segment drive shafts

Note: 2,000 L systems include two‑piece drive shafts. Before starting to load a drive shaft, verify that the drive shaft serial numbers match on both shaft segments.

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.

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Chapter 3 | Operating information

Operating information

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

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Chapter 3 | Operating information

3.1 General system operating information

3.1.1 BPC preparation

Each outer support container is designed for a speciﬁc BioProcess Container (BPC). Conﬁrm that the correct volume and type of BPC is being used for the corresponding volume outer support container. 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 Single‑Use Bioreactor (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. Actual working volumes should not exceed the indicated rated working volumes by more than 10%.

CAUTION: Operating 2:1 S.U.B.s at working volumes less than 50% of the rated volume without consultation from Thermo Fisher Scientiﬁc engineers 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.

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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.

The standard BPC is designed with special spargers that produce very efﬁcient mass transfer of oxygen and typically will require much less gas inﬂow than conventional spargers. Gas inﬂow should only be limited to prevent foam generation and excessive pressure within the BPC. Gas ﬂow rates supplied as overlay should also be reduced as much as possible or eliminated, which will minimize both liquid evaporation and demand on the exhaust ﬁlter. Ideally, this will reduce the likelihood that gas inﬂows will exceed gas outﬂow of the system and reduce the occurrence of foam in the headspace that may plug the exhaust ﬁlter. For more information, refer to the "Operating pressure" section on the previous page, and section 3.6.8 of this guide.

Aseptic connections

The most commonly recommended process for making connections to 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.

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Chapter 3 | Operating information

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 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 50%

liquid volume or greater. Batch temperature should not exceed

40°C.

Table 3.1. Approximate heating times for 2:1 S.U.B. systems. Ambient temperature of 25°C.

Liquid batch

System

50 L electric 25–50 L 456 N /A 18.2–9.2 W/L 5°C 37°C N/A–4.3 hr

50 L jacketed 25–50 L 2800 TF2500 112– 5 6 W/ L 5°C 37°C 0.8–1.1 hr

100 L electric 50–100 L 865 N/A 17.3–8.7 W/L 5°C 37°C N/A– 4.9 hr

100 L jacketed 50–100 L 2800 TF2500 56–28 W/L 5°C 37°C 1.2 –1.9 hr

250 L electric 125–250 L 1358 N/A 10.9–5.4 W/L 5°C 37°C N/A –7. 5 h r

250 L jacketed 125–250 L 2800 TF5000 22.4–11.2 W/L 5°C 37°C 2.1– 3.1 h r

500 L jacketed 250–500 L 6100 TF10000 24.4–12.2 W/L 5°C 37°C 1.5–2.6 hr

1,000 L jacketed 500–1,000 L 22500 TF24000 45 –22.5 W/L 5°C 37°C 1.3 –1.8 hr

2,000 L jacketed 1,000–2,000 L 22500 TF24000 22.5–11.3 W/L 5°C 37°C 2–2.8 hr

volume (half

[min.]–full)

Watts TCU

TCU watts/L

(half–full)

Initial liquid temp.

Liquid target

temp.

Time

(half–full)

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Chapter 3 | Operating information

Protective earth grounding for 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 electrical box enclosures

The agitation control interface utilizes an LED 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 50%. Stirring speed is adjusted using the up and down arrows on the agitation keypad interface on the control panel, or using the settings on an integrated third‑party controller. Note: 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 control panel. For more information, see the illustrations in the control panel detail in section 4.3.

Circuit protection for 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.

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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 Figures 2.2 and 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.

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.

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Chapter 3 | Operating information

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. 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 (Figure 3.3),

close the door, and close the clamp.

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Chapter 3 | Operating information

Figure 3.1. BPC removed from protective polybags.

Figure 3.2. BPC loading.

Figure 3.3. Bearing port insertion.

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).

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.

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Figure 3.5. Sparge line setup.

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 system in use.

Figure 3.8. Incoming line connection to inlet ports.

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).

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Chapter 3 | Operating information

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 standard liters per minute (slpm) or 0.03 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 0.03 bar (0.5 psi) within the BPC or it could fail. For reference, the BPC will appear to be tight at 0.007 bar (0.1 psi). See Tables 3.7 and 3.8 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.

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.

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Chapter 3 | Operating information

Figure 3.10. 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.

15. Connect the media ground clip to the stainless steel insert in the

sample line on the BPC. This grounds the media inside the BPC and helps eliminate electrostatic charge (Figure 3.12).

Ground clip connection

Figure 3.12. Media ground clip connection.

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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 cap

Hollow passthrough

Safety cover

Drive shaft

Drive shaft head

Motor drive keyway

Latch pin

Motor

Figure 3.13. Motor and mixing assembly.

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).

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Chapter 3 | Operating information

Figure 3.14. Removing motor cap.

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 a n d 3.16 ).

Figure 3.15. Loading drive shaft.

Figure 3.16. Twisting drive shaft to aid insertion.

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Chapter 3 | Operating information

• 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. Engaging impeller.

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.

Figure 3.18. Drive shaft head aligned.

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Chapter 3 | Operating information

• 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 is calibrated at the factory at 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).

Figure 3.21. Replacing and latching cover.

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Chapter 3 | Operating information

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. Note: 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.

4. Attach the overlay sparge line and any other lines to the cable management system, if available (Figure 3.23). Then, position and close a bar clamp on the bottom drain line as close as possible to the BPC port (Figure 3.24).

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Chapter 3 | Operating information

Figure 3.23. Optional cable management system on a S.U.B. unit.

Figure 3.24. Bar clamp installation.

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

6. Insert the resistance temperature detector (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 counter‑clockwise to aid insertion.

• Secure by twisting the luer lock collar, if provided. The thermowell will stretch slightly when the RTD is seated.

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Chapter 3 | Operating information

Figure 3.25. Sensor insertion.

7. Optional: Connect a pressure sensor to the 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.

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

Checkpoints prior to BPC loading

9 The correct volume BPC is being used for the corresponding

volume outer support container.

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.

Figure 3.26. Securing sensor.

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).

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Chapter 3 | Operating information

Figure 3.27. Opening the bioreactor door.

Figure 3.28. Close-up of bearing receiver clamp.

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 removed from protective polybags.

Figure 3.30. Bearing port orientation.

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.

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Chapter 3 | Operating information

Figure 3.31. Line sets on edge of tank.

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.

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 3.33. Door clamped shut.

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Chapter 3 | Operating information

Figure 3.34. Sparge line insertion.

Figure 3.35. Sparge line extension.

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 insertion.

Figure 3.37. Drain/sampling line extension.

8. Connect the pressure transducer to the monitor. After the display has stabilized, tare the monitor. Note: The monitor should be allowed to warm up for 30 minutes before taring. Verify that the monitor reads zero.

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Chapter 3 | Operating information

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 be less than 25 slpm or 0.2 bar (3 psi).

• 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 through 13 can be completed while the BPC is ﬁlling

with air.

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

EXCEED 0.03 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 Tables 3.7 and 3.8 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.

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). 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 2 hanging tabs to the pins to assist in aligning the probe belt, drain port, and spargers.

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Chapter 3 | Operating information

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 the 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).

Figure 3.40. Removing protective packaging.

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Chapter 3 | Operating information

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. with the use of a 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.

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.

Motor cap

Hollow passthrough

Safety cover

Drive shaft

Drive shaft head

Motor drive keyway

Latch pin

Motor

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).

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Chapter 3 | Operating information

Figure 3.42. Latch pin removal. Figure 3.43. Opening safety cover.

Figure 3.44. Cap removal.

2. Verify that the two or three 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. Each time drive shafts are assembled and used,

operators must verify that the segments have matching serial numbers.

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.

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Chapter 3 | Operating information

Figure 3.45. Inserting lower section.

Figure 3.47. Segment assembly.

Figure 3.46. Latch pin in shaft.

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.

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Chapter 3 | Operating information

Figure 3.48. Tightening of shaft connections.

5. Load the partially‑assembled drive shaft through the hollow pass‑ through and hold it in position with the latch pin. Obtain the upper segment of the drive shaft and assemble it to the middle segment in the manner described previously.

6. Using two hands, carefully guide the assembled 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.

• 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.

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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 150 in‑lb.

Figure 3.51. Tightening motor cap with wrenches.

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Chapter 3 | Operating information

9. Verify that the wrenches have been 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, position the line clamp as close as possible to the BPC port, and close the clamp. Use a cable tie around the clamp to ensure it does not open.

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

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Chapter 3 | Operating information

Figure 3.54. Aseptic connector port alignment.

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

6. Insert a resistance temperature detector (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. Inserting a temperature sensor.

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Figure 3.56. Securing the temperature sensor.

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 passthrough 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.

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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 BioProcess Container (BPC) is being used for

the corresponding volume outer support container.

9 Three operators are available for 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. Use an elevated platform to open the clamp on the bearing port receiver located below the motor. Lower the hoist lifting frame to a position just above the top of the rear door by using the pneumatic control lever (near the rear door).

3. 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.

Figure 3.58. Removing the BPC from polybags.

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Chapter 3 | Operating information

4. 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 lifted by the hoist.

Figure 3.59. Loading the BPC.

5. Using the rear or front door for access, connect the retainer hooks on the hoist to the top of the BPC via the hanging tabs, starting with the furthermost two tabs (Figure 3.60). Finish with the closest two tabs (Figure 3.61).

Figure 3.60. Connecting furthermost retainer hooks.

Figure 3.61. Connecting closest retainer hooks.

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Chapter 3 | Operating information

6. Raise the BPC using the pneumatic lift. One operator should observe from above while another operator controls the lifting valve at ground level (Figure 3.62).

CAUTION: While operating the 2,000 L bag hoist watch for excess stress on the BPC during lifting. Reposition the BPC as necessary to avoid tearing the BPC hanging tabs.

Figure 3.62. Operating the pnuematic lift control to raise the bag hoist.

7. Raise the hoist until the lift reaches full stroke. Once lifting stops, the top‑level operator should hold the hoist frame and apply minimal lifting force. This will assist the lifting device to pull in any remaining slack in the cable, and ensure the lift device has been fully raised. Place the valve in the "stop" position.

8. 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 3.63), load the BPC bearing port into the receiver (Figure 3.64), close the bearing assembly door, and latch it (Figure 3.65).

Figure 3.63. Removing the cap. Figure 3.64. Bearing port loading. Figure 3.65. Closing/latching door.

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Chapter 3 | Operating information

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

10. Open the tubing set polybag and connect the pressure transducer to the monitor. After the display has stabilized, tare the monitor. Note: Allow the monitor to warm up for 30 minutes and connect the sensor 10 minutes before taring. Verify that the monitor reads zero.

11. 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 3.66). Carefully center the ﬁlter housing, allowing the clip to secure it near the hose barb connections.

Figure 3.66. Clipping ﬁlter to holder.

• Ensure that the routing of the exhaust tubing is not likely to

become kinked.

• Place the vent heaters around each ﬁlter (Figure 3.67), verifying

that the snap retainers are secured. Position the power leads to avoid interfering with the vent holder brackets.

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Chapter 3 | Operating information

Figure 3.67. Installing heaters.

• Raise and rotate the vent holder bracket as needed (Figure 3.68).

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 3.68. Raising the vent holder bracket.

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

the controllers.

• Inspect the controller 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.

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Chapter 3 | Operating information

12. Fill the BPC with air via the DHS and overlay gas inlet line.

13. Clamp the drilled hole sparge and exhaust lines so that the air supplied by the overlay gas inlet line ﬂows directly into the BPC (Figure 3.69). Note: Remove the clamp prior to sparging.

Figure 3.69. 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. Allow the container to ﬁll to greater

than half volume. This typically takes less than 20 minutes.

• Steps 15 through 18 can be completed while the BPC is ﬁlling

with air.

Note: Air pressure to the overlay gas line on the BPC should be less than 100 slpm or 0.34 bar (5 psi).

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

EXCEED 0.03 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.

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Chapter 3 | Operating information

15. Feed the probe belt, sample line, and the subsurface addition lines through the front access door (Figure 3.70).

Figure 3.70. Feeding lines through the front access door.

16. Remove the sparge lines from the polybags and the 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 3.71).

Figure 3.71. Clamping bottom drain line.

17. The center insert on the tank ﬂoor provides the port locations for both the bottom drain (Figure 3.72) and the gas lines for the open pipe or drilled hole sparger. Guide the sparger inlet line and ﬁlter through the bottom cutout in the tank (Figure 3.73) to provide access for loading the porous frit sparger gas line (Figure 3.74). To remove the bottom cutout, lift and rotate it in a counterclockwise direction.

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Chapter 3 | Operating information

Figure 3.72. Drain line support.

Figure 3.74. Porous frit sparger line loading.

Figure 3.73. Macro sparger line support.

18. Three cutout holes are provided in the tank for porous frit spargers. These holes are located furthest from the tank center line and align with the inside edge of the access cover door (Figure 3.75).

Figure 3.75. Cutouts for porous frit spargers.

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Chapter 3 | Operating information

Sparger notes:

• Air pressure to the spargers on the BPC should not exceed

0.55 bar (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.

19. Attach all of the hanging tabs to help position the ports. Secure the BPC by attaching the tabs on the bottom of the BPC onto the position tab pins (Figures 3.76 and 3.77). 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.

Position tab pin

Figure 3.76. Pulling the container tab toward the pin.

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

Figure 3.77. Securing the container tab on the pin.

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Chapter 3 | Operating information

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 prevents 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 BPC 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 3.78 and 3.79 show both the cart assembly and side‑mounted (2,000 L systems only) condenser system options.

Condenser disposables Condenser hardware

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 3.78. 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

Complete condenser system

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Chapter 3 | Operating information

Figure 3.79. Side-mounted condenser system option for 2,000 L S.U.B.s.

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 2:1 Single‑Use Bioreactor Validation Guide (DOC0016) for details.

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Chapter 3 | Operating information

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 5 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 that no foam is present in the exhaust stream.

Table 3.2. Condenser system overview.

S.U.B. system

2 each 254 mm (10 in.) vents

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

Run

duration

Maximum combined ﬂow

rate recommended

Resulting

safety factor

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 a 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.

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2,000L S.U.B. BPC - Pressure Trending

Overlay Pressure (psi)

Air Sparge (lpm)

Direct Sparge Gas Loading, 2,000 L DI Water @ 40°C, Vent Heater @ 60°C

0.25

0.20

0.15

0.10

0.05

20 30 40

60 70 80 90

Single Meissner 10” Vent (0.2 micron PVDF)

Chapter 3 | Operating information

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

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. This tubing diameter will allow condensate to return to the S.U.B. at total off‑gas ﬂow rates up to 30 slpm, assuming that 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, 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;

12.7 mm (0.5 in.) inner diameter (ID) tubing is typically deemed too small for the 2,000 L S.U.B.

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Tubing I.D. Recommendation (inches)

Gas Vapor Velocity (m/s)

Flow (lpm)

Flow Rate (lpm) Versus Vapor Exit Velocity (m/s) - 0.61 m/s Velocity Threshold

0.5

1.0

1.5

2.0

2.5

20 30 40

0.63 0.75 1.00 Threshold

60 70 80 90 100

Chapter 3 | Operating information

Graph 3.2. Tubing inner diameter recommendation.

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 on 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 the event that issues arise in process).

™

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Chapter 3 | Operating information

Graph 3.3. Off-gassing management guidelines.

Condenser system setup

1. Remove the reservoir cap of the chiller and add the appropriate type and volume of ﬂuid 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.

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Chapter 3 | Operating information

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.

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. (Figure 3.80).

Figure 3.80. Removing condenser from packaging.

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Chapter 3 | Operating information

2. The operator at the upper position should move to ground level, open both doors on the chiller plate (Figure 3.81), and load the condenser BPC from the front, keeping the BPC in a saddle bag shape. Allow the vents to hang freely.

Figure 3.81. Opening the chiller plate doors.

3. Route the gas inlet lines around and behind the vent holders. Inspect both lines to ensure they are connected to the S.U.B. and are not twisted or kinked (Figure 3.82). Adjust as needed.

Figure 3.82. Routing gas inlet lines.

4. The second operator should hold the vent ﬁlters and place them into the vent ﬁlter holders above the chill plate (Figure 3.83).

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Chapter 3 | Operating information

Figure 3.83. Placing vent ﬁlters.

5. The ﬁrst operator should use the Velcro straps to secure the ﬁlters in position (Figure 3.84). Then use the grasping tabs to position the container using the two lower button pins on each side of the chiller plate (Figure 3.85).

Figure 3.84. Securing vent ﬁlters. Figure 3.85. Positioning container.

6. Close the clear side doors while carefully manipulating ﬁrst the gas inlet line and then the gas outlet line (Figure 3.86) to clear the doors as each is closed and latched (Figure 3.87).

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Chapter 3 | Operating information

Figure 3.86. Moving gas lines.

Figure 3.87. Latching door.

7. As the doors are closed and latched, the second operator should route the gas inlet lines into the clips behind the vents (Figure 3.88).

Figure 3.88. Clipping gas lines into place.

8. 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 3.89).

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Chapter 3 | Operating information

Figure 3.89. Loading pump tubing.

9. Start the pump by pushing the red button (Figure 3.90). 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 3.90. Starting the pump.

10. 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.

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Chapter 3 | Operating information

3.4.3 Drive shaft insertion for 2,000 L systems

The drive shaft is constructed by assembling two quick‑connect segments. These segments must be assembled and inserted in sections. Operators should be elevated (i.e. with the use of a ladder) to 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.

Figure 3.91 illustrates the components of the motor and mixing assembly. The parts labeled on the ﬁgure will be referenced throughout the drive shaft insertion process.

Motor cap

Hollow passthrough

Safety cover

Drive shaft

Drive shaft head

Motor drive keyway

Latch pin

Motor

Figure 3.91. Motor and mixing assembly.

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Chapter 3 | Operating information

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 3.92), opening the safety cover (Figure 3.93), and removing the threaded cap by turning it counterclockwise. Use the wrench and spanner provided, if necessary.

Figure 3.92. Removing latch pin. Figure 3.93. Opening safety cover.

4. Verify that both 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. The segments will be referred to as the upper (segment with the drive shaft head) and lower (segment with the square end). Important notes: Each time drive shafts are assembled and used in the S.U.B., operators must verify that the segments have matching serial numbers. 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 3.94). Once inserted, slide the latch pin from the mixing assembly into the shaft to prevent it from falling into the impeller sleeve (tube) (Figure 3.95).

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Chapter 3 | Operating information

Figure 3.94. Insertion of the lower section of the drive shaft.

Figure 3.95. Use of the latch pin.

6. To connect two shafts together, depress the button on the female side (Figure 3.96) and slide the sleeve back. This will expose a red ring underneath the sleeve (Figure 3.97). This is a visual indicator that the sleeve is not in a locked position.

Figure 3.96. Button used for connection.

Figure 3.97. Sliding sleeve exposing red "not locked" indicator.

7. Obtain the upper section of the drive shaft. Place the female side of the quick connect over the male end of the lower section (Figure 3.98). The connection is fully seated when the red indicator ring (Figure 3.99) on the male end is no longer exposed.

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Chapter 3 | Operating information

Figure 3.98. Quick connection.

Figure 3.99. Red indicator ring exposed.

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 3.100).

Note: When fully connected, no red coloring should be visible.

Figure 3.100. Sliding the sleeve into place.

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

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Chapter 3 | Operating information

10. Using two hands, carefully guide the completed drive shaft into the BPC using a slight back and forth twisting motion, or a counter‑ clockwise rotation (Figure 3.101). Do not push the drive shaft

straight in.

Note: Figures 3.101 and 3.102 show a drive shaft with a white shaft

head. All 2,000 L S.U.B.s use a new, longer drive shaft with a black drive shaft head.

Figure 3.101. Insertion of the drive shaft.

Figure 3.102. Engaging the bearing port.

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

to engage the impeller.

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

to engage the bearing assembly (Figure 3.102, above).

• When 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.

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 3.103 illustrates a drive shaft that is completely 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 10 before replacing the cap.

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Chapter 3 | Operating information

Drive shaft end

Figure 3.103. Drive shaft fully inserted into the impeller.

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 3.104).

Note: The torque wrench is a standard 10 mm (3/8 in.) square drive, and is calibrated at the factory at 150 in‑lb.

Figure 3.104. Tightening cap.

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.

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Chapter 3 | Operating information

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, porous frit, 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 3.105). This will be the lower cutout if your system is supplied with two horizontal slots. Note: The latest style of hardware has a third opening.

Figure 3.105. Aseptic port alignment.

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 3.106).

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