ThermoFisher DOC0031 User Manual

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HyPerforma Single-Use Fermentor (S.U.F.) User’s Guide

DOC0031 • Revision G February 2021

Contents

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

Chapter 1 Single-Use Fermentor (S.U.F.) overview 10

1.1 Introduction 11

1.2 Hardware characteristics 14

1.2.1 System features 14

1.2.2 Agitation 14

1.2.3 Control system 14

1.2.4 Exhaust management system 14

1.2.5 Exhaust vent ﬁlter heater 14

1.2.6 Temperature 15

1.2.7 Heating performance 15

1.2.8 External control 15

1.2.9 Load cells 15

1.3 End user and third-party supplied components 16

1.3.1 pH and dissolved oxygen (DO) probes 16

1.3.2 Single-use probes 16

1.3.3 Controllers 17

1.4 BioProcess Container characteristics 18

1.4.1 Single-Use Fermentor BPC features 18

1.4.2 Operating pressure 18

1.4.3 Working volume 18

1.4.4 Draining and harvest 19

1.4.5 Aeration 19

1.4.6 Aseptic connections 19

1.4.7 Sampling 19

1.4.8 BPC features 20

1.5 Additional system components 22

1.5.1 Probe integration 22

1.5.2 Accessories 22

Contents

Chapter 2 Installation and setup 24

2.1 Initial installation preparation 25

2.1.1 Hardware shipment and setup 25

2.2 Site preparation 25

2.2.1 Electrical connections for units with an electrical control box 25

2.2.2 Outer support container preparation 26

2.2.3 Electrical preparation for systems with electrical control boxes 26

2.2.4 Load cell preparation 27

2.3 Installation 28

2.3.1 Exhaust ﬁlter bracket setup 30

2.3.2 Condenser unit setup (when present) 31

2.3.3 TCU/chiller connection to condenser unit (when present) 32

2.3.4 Exhaust ﬁlter pinch clamp setup (when present) 33

2.3.5 Optional cable management system setup 35

2.3.6 Air line preparation 35

2.3.7 Drilled hole sparger line(s) 35

Chapter 3 Operating information 36

3.1 General system operating information 37

3.1.1 BPC preparation and loading 37

3.1.2 BPC handling instructions 37

3.1.3 BPC operating information 37

3.1.4 Hardware operating information 39

3.2 BPC and drive shaft loading for 30 L units 43

3.2.1 Initial installation steps for 30 L units 43

3.2.2 Drive shaft insertion for 30 L units 51

3.2.3 Final installation steps for 30 L units 54

3.3 BPC and drive shaft loading for 300 L units 56

3.3.1 Initial installation steps for 300 L units 56

3.3.2 Drive shaft insertion for 300 L units 64

3.3.3 Final installation steps for 300 L units 69

3.4 Exhaust system 72

3.4.1 Exhaust system functional overview 72

3.4.2 When to use the condenser 73

3.4.3 Condensers and reserve vent ﬁlters 74

Contents

3.4.4 Exhaust system setup 74

3.4.5 Exhaust system setup 75

3.5 Probe assembly 79

3.5.1 Preparation and sterilization 79

3.5.2 Making CPC AseptiQuik connections 80

3.5.3 Probe insertion 83

3.5.4 Probe calibration 85

3.5.5 Kleenpak connection instructions 85

3.6 Microbial cell culture operating instructions 95

3.6.1 Operating conditions for cell culture applications 95

3.6.2 Manual operation of drilled hole spargers 96

3.6.3 Checkpoints prior to media ﬁll 97

3.6.4 Media ﬁll 97

3.6.5 Agitation for units with electrical control boxes 98

3.6.6 Temperature control 99

3.6.7 pH probe calibration 100

3.6.8 DO probe calibration 100

3.6.9 Checkpoints prior to inoculation 100

3.6.10 Cell inoculation 100

3.6.11 In-process checkpoints 101

3.6.12 Dispense and harvest 101

3.6.13 S.U.F. shutdown and post-use BPC disposal 102

3.6.14 Preparation for the next run 103

3.7 BPC sampling 104

3.7.1 Aseptic sampling 104

3.7.2 Sampling with sterile manifold 105

3.8 Veriﬁcation procedures 107

3.8.1 Mixer speed veriﬁcation 107

3.8.2 Temperature controller veriﬁcation 107

3.8.3 Pressure monitor veriﬁcation (when present) 107

3.8.4 Load cell veriﬁcation (when present) 107

Chapter 4 Speciﬁcations and parts information 108

4.1 Hardware features and dimensions 109

4.1.1 30 L S.U.F. design features 109

4.1.2 300 L S.U.F. design features 110

4.1.3 30 L S.U.F. dimensions and ordering information 111

4.1.4 300 L S.U.F. dimensions and ordering information 112

Contents

4.2 30 L and 300 L S.U.F. hardware speciﬁcations 113

4.3 Conﬁgurable hardware options 115

4.4 BPC features and speciﬁcations 116

4.4.1 Standard 30 L S.U.F. BPC 116

4.4.2 Standard 300 L S.U.F. BPC 118

4.5 BPC options and packaging information 120

4.6 Accessories and options speciﬁcations 121

4.6.1 Load cells 121

4.6.2 Cable management system 124

4.6.3 Vent ﬁlter heaters 125

4.6.4 Exhaust system 126

4.6.5 Miscellaneous items 129

Chapter 5 Maintenance and troubleshooting 132

5.1 Maintenance 133

5.1.1 Routine maintenance 133

5.1.2 Preventive maintenance schedule 133

5.2 Troubleshooting and frequently asked questions 135

5.2.1 Hardware operation issues 135

5.2.2 Cell culture operation issues 137

5.2.3 Probe and connector issues 140

5.2.4 BPC and sparging issues 141

Chapter 6 General ordering information 142

6.1 Ordering instructions 143

6.2 Ordering/support contact information 143

6.3 Technical support 144

Appendices Appendix A—Installation of female electrical receptacle

for units with AC motors and electrical boxes 145

Appendix B—Mettler Toledo IND331 Display load cell

calibration instructions 148

Appendix C—Pre-fermentation run checklist 150

Warnings, safety, and warranty information

Thank you for purchasing the Thermo Scientiﬁc™ HyPerforma™ Single-Use Fermentor (S.U.F.). 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 this equipment.

The Thermo Scientiﬁc HyPerforma Single-Use Fermentor (S.U.F.) is designed to be operated under traditional microbial cell culture conditions. A general understanding of Fermentor systems and their operation is important prior to using the system for the ﬁrst time. Read and understand user’s guide before operating; failure to do so could result in injury. Proper procedures for the disposal of Single-Use

BioProcess Containers (BPCs) should be followed, depending on the culture in use. See the appropriate section of this guide.

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: Hazardous voltage inside.

Electrical components are required for the proper function of the S.U.F. 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 by trained personnel only. Thermo Fisher Scientiﬁc recommends using standard lockout procedures when working on electrical components. The main breaker on the electrical control panel may be locked out.

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Warnings, safety, and warranty information

WARNING: Static electricity may build up in BPCs.

• BPCs may act as insulators for electrostatic charge. If electrostatic

• Where applicable, a product contact stainless steel coupler may be

WARNING: Rotating parts—entanglement hazard.

Rotating and moving parts can cause injury. Keep hands away from moving parts.

• Do not operate this equipment unless the supplied guarding is in

• It is the responsibility of the end user to assess this equipment

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.

grounded to the frame to dissipate electrostatic build up from the material within a BPC. It is good practice to dissipate electrostatic build up by grounding all BPCs prior to coming in contact with them. When working with BPCs, the use of nonconductive materials, such as nonconductive gloves, is recommended.

place and properly functioning.

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 mounting the motor to the 300 L S.U.F. during setup, 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.

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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: The Thermo Scientiﬁc HyPerforma S.U.F. may not be installed in a potentially explosive atmosphere as set forth in the applicable EU ATEX Directive.

WARNING: Burst hazard.

The S.U.F. 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.

• Do not exceed 0.03 bar (0.5 psi) pressure within the BPC

• Do not exceed 0.34 bar (5 psi) pressure in the section of BPC

• Make sure the vent ﬁlter is properly positioned and working

• Maintain a minimum gas ﬂow of 0.1 vvm or 0.01 psi in the BPC

tubing between the inlet ﬁlter and the BPC

properly

while the impeller is spinning

Protective earth grounding

Protective earth grounding must be veriﬁed prior to plugging the S.U.F. 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

Water jacket vessel information

S.U.F. 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 150 psig (1 MPa) operating pressure. For the utmost safety it is recommended that the water jacketed S.U.F. be operated at 75 psig or less.

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Warnings, safety, and warranty information

Note: The S.U.F. BPC operating limits for temperature are 5°C to 40°C. The internal pressure should not exceed .003 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.

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.F. not be plugged into a GFCI outlet.

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

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Warnings, safety, and warranty information

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.

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.

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Warnings, safety, and warranty information

Seismic guidance

The buyer of the equipment is responsible for ensuring that countryspeciﬁc 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. Upon request, Thermo Fisher Scientiﬁc can assist with establishing compliant seismic anchoring and tethering designs for purchased equipment based on building and country codes, 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.

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How to use this guide

Rev. Date Section Change made Author

A.0 07/ 2 014 -- Initial release S. Jelus

A 11/ 2 0 16 -- Reformatted E. Hale

A 11/ 2 0 16 3.1.4, 4.2 Updated maximum temperature setpoint J. Brown/E. Hale

A 11/ 2 0 16 3.5.2 Added CPC AseptiQuik connection instructions E. Hale

A 11/ 2 0 16 3.5.5 Removed references to sterility E. Hale

A 11/ 2 0 16 4.2 Updated nominal tip speed E. Hale

A 11/ 2 0 16 Appendix D Added Appendix D (Pre-fermentation run checklist) J. Brown/E. Hale

B 04/2017 4.2

B 04/2017 4.4.1

B 05/2017

B 05/2017 2.3, 4.1, 4.6

C 09/2017

C 09/2017 -- Changed values from “psi (bar)” to “bar (psi)” E. Hale

C 09/2017 -- Updated various aspects of formatting E. Hale

C 09/2017

C 09/2017 4.2 Added “Noise level” to speciﬁcations table E. Hale

C 11/ 2 0 17 Chapter 4 Removed hardware, BPC part numbers E. Hale

C 01/2018 -- Updated warning symbols E. Hale

How to use this guide

Scope of this publication

This user’s guide contains information about the standard Thermo Scientiﬁc S.U.F. 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

Warnings and

Safety

Warnings, safety,

and warranty

information

How to use this

guide

Added Minimum Ceiling Height Requirement to 30 and 300 L speciﬁcations

Replaced “+/- 0.75% of full scale” with “+/- 5 rpm” in Motor Speed speciﬁcation ﬁeld

Corrected part number for standard 30 L BPC with two 5 in. exhaust ﬁlters

Added potentially explosive atmosphere (ATEX) warning E. Hale

Updated 30 L S.U.F. drawings to show the current motor mount

Added warranty and usage information E. Hale

Added “Abbreviations/acronyms” section E. Hale

E. Hale

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How to use this guide

Rev. Date Section Change made Author

D 11/2 018

D 11/2 018 Various Removed references to metal probe clips E. Hale

D 11/2 018 2.2.3

D 11/2 018 3.6.5, 4.6.1 Updated Figures 3.104 and 4.12 E. Hale

D 11/2 018 Appendices Removed Appendix B (AC-Tech calibration) E. Hale

D 11/2 018

D 12 / 2018 3.1.3, 3.6.5 Edited sentence (3.1.3) and reworded step #2 (3.6.5) E. Hale

Warnings, safety,

and warranty

information

How to use this

guide

Warnings, safety,

and warranty

information

Added seismic guidance E. Hale

Changed “Input into Thermo Scientiﬁc publications” to “Questions about this publication”

Updated Figure 2.1, showing the interior of S.U.F. E-Boxes, and removed Figure 2.2

Added emphasis to “Electrical connections” section E. Hale

E. Hale

D 12 / 2018 4.2

E 10/ 2019 -- Minor revisions T. Golightly

E 10/ 2019 4.2 Updated cart length dimension on Table 4.2 T. Golightly

Warning, safety,

F 06/2020

G 11/2020 --

G 11/2020 4.1.3 Updated Figure 4.5 with new cart dimentions T. Golightly

G 11/2020 4.2

G 11/2020 1.3.1

and warranty

information

Added tolerance (± 5 rpm) to “Motor speed” in speciﬁcations

Added Tipping hazard warning and symbol C. Jones

Minor updates to the document to meet current document standards

Corrected the overall width, length, height and dry and wet skid weights in Table 4.2

Updated the "Finesse pH and DO" sensors to "Hamilton pH and DO" sensors in Table 1.1

E. Hale

T. Golightly

T. Golightly/E. Hale

T. Golightly

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.

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How to use this guide

Related documents

If you are missing any of the related publications listed below, please contact your local sales representative.

Document Document number

Thermo Scientiﬁc HyPerforma S.U.F. Validation Guide DOC0032

Thermo Scientiﬁc HyPerforma S.U.F. Packing and Unpacking Instructions

DOC0052

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 ID Inner diameter IEC International Electrical Code OD Outer diameter PED Pressure Equipment Directive PID Proportional integral derivative P/V Power input to volume rpm Revolutions per minute RTD Resistance temperature detector slpm Standard liters per minute STR Stirred tank reactor S.U.F. Single-Use Fermentor TCU Temperature control unit VFD Variable frequency drive

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Chapter 1 | S.U.F. overview

Single-Use Fermentor (S.U.F.) overview

Chapter contents

1.1 Introduction

1.2 Hardware characteristics

1.3 End user and third-party supplied components

1.4 BioProcess Container characteristics

1.5 Additional system components

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Chapter 1 | S.U.F. overview

1.1. Introduction

The Thermo Scientiﬁc Single-Use Fermentor (S.U.F.) has been designed to be a single-use alternative to conventional stirred tank aerobic fermentors currently utilized in microbial cell culture applications. Based on years of accepted stirred tank reactor (STR) design, the S.U.F. 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.F. an attractive alternative to its stainless steel counterpart. Critical design parameters such as height to diameter ratios, mixer design and location, and typical control system interfaces are consistent with traditional fermentation platforms. S.U.F. BioProcess Containers (BPCs) are supplied sterilized by irradiation and therefore do not require any facility hook-ups for sterilization or cleaning. A key element in 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 microbial cell culture runs over traditional reusable systems.

S.U.F. systems consist of the following primary components:

• Outer support container with standard water jacket heating system

• S.U.F. BioProcess Container (BPC)

• Electrical control panel (E-Box) for agitation

• Direct drive agitation mixing assembly with an AC motor, drive

shaft, and impeller

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Chapter 1 | S.U.F. overview

Figure 1.1. 300 L S.U.F. with electrical control panel, condenser, cable management system, and bottle management system.

The Thermo Scientiﬁc S.U.F. 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 HMI (human-machine interface) screens. Controls for agitation are integrated into the S.U.F., with temperature, pH/DO probes and controls being supplied by the user or a third-party integrator. Water jacketed systems require a temperature control unit (TCU) selected and supplied by the end user or by Thermo Scientiﬁc.

The stainless steel outer support container is engineered and fabricated to fully support each S.U.F. BPC and allow easy access for operation. The outer support container is mounted to a portable cart base and includes the mixing drive, water jacket for heating and cooling, and optional controllers for mixing. The drive shaft is detachable and reusable and is inserted into the BPC through the motor assembly and into the bearing port. Load cells are offered as an option.

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Chapter 1 | S.U.F. overview

The BPC includes the impeller assembly, sparger, vent ﬁlter inlet/outlet ports, probe integration ports and ﬁlling, dispensing, and sampling ports. 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 or sterilization validations are required as the BPC is provided gamma irradiated. Innovative, proprietary technology allows for the integration of the mixing shaft, 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 S.U.F. BPC through universal ports.

This user’s guide covers the setup, operation, maintenance, and troubleshooting of 30 and 300 L S.U.F. systems.

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Chapter 1 | S.U.F. overview

1.2. Hardware characteristics

1.2.1. System features

The S.U.F. is designed for system mobility and easy integration while utilizing a straightforward operator interface. Hardware drawings and speciﬁcation tables speciﬁc to volume can be found in Chapter 4 of this guide. The following topics include general descriptions of the hardware components of the S.U.F.

1.2.2. Agitation

If your system uses 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 revolution 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. Your system may also be integrated and managed by a third-party controller.

1.2.3. Control system

The S.U.F. is designed to integrate with existing control systems in their many conﬁgurations. The S.U.F. control system supplied with the Thermo Scientiﬁc E-Box enclosure manages the agitation process and monitors the pressure inside the BPC. 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 thirdparty integrator.

1.2.4. Exhaust management system

A condenser system is recommended and is available as optional hardware. It cools exhaust gases and re-circulates the condensate into the fermentor.

1.2.5. Exhaust vent ﬁlter heater

An optional exhaust vent ﬁlter heater is available for increased longevity of the exhaust ﬁlter on the S.U.F. BPC. Heating the ﬁlter sufﬁciently to eliminate the formation of condensation is an effective means of reducing the risk of fouling of the ﬁlter membrane. The heater is factory preset to operate between 40–55°C, ± 3 degrees. Temperature settings above 60°C are not recommended.

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Chapter 1 | S.U.F. overview

1.2.6. Temperature

The S.U.F. can be operated within the temperature range from 5–55°C. The process temperature is measured by means of a supplied RTD (pt-100) that is inserted into the thermowell of the S.U.F. BPC. Water jacketed system temperature control is maintained through the TCU or a third-party controller.

1. 2 .7. Heating performance

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

1.2.8. External control

Users can choose to bypass the mixing speed control and display pressure systems provided with the S.U.F. and utilize existing controllers. Refer to section 3.6 for more details.

1.2.9. Load cells

Load cells, used to determine the weight of the contents of a S.U.F., are optional. Load cell kits 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 data interfaces.

Load cells arrive uncalibrated. It is recommended that the load cell manufacturer or a qualiﬁed technician calibrate these systems on site.

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Chapter 1 | S.U.F. overview

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

Probe lengths (from O-ring to tip) must not exceed 235 mm (9.25 in.)

1.3. End user and third-party supplied

components

1.3.1. pH and DO probes

The following table shows the length and diameter requirements for traditional sensors (probes) that can be integrated into the S.U.F. 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.F.

O-ring to probe tip

Sensor Inserted

Probe Part number Diameter

AppliSens DO Z010023525 12 mm (0.47 in.) 13.5 PG

AppliSens pH Z001023551 12 mm (0.47 in.) 13.5 PG

Mettler Toledo DO

Mettler Toledo pH

Broadley-James DO D140-B220-PT-D9 12 mm (0.47 in.) 13.5 PG

Broadley-James pH F-635-B225-DH 12 mm (0.47 in.) 13.5 PG

Hamilton DO 237542 12 mm (0.47 in.) 13.5 PG

Hamilton pH 238633-2543 12 mm (0.47 in.) 13.5 PG

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

InPRO 6800/12/220, PN 52200966

405-DPAS-SCK8S/225, PN 104054481IG

12 mm (0.47 in.) 13.5 PG

Thread

type

Length Length

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.)

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.)

1.3.2. Single-use probes

Mettler Toledo single-use pH and DO sensors are supported, and BPCs may be ordered with single-use sensors as fully integrated components.

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Chapter 1 | S.U.F. overview

1.3.3. Controllers

Thermo Scientiﬁc operates with an open architecture approach to the integration of controls. Our industry-leading S.U.F. can be 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 Thermo Scientiﬁc S.U.F.

S.U.F. units may be provided with integrated controls, pump towers, a control monitor, and advanced features, such as data logging, multiple S.U.F. 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 Scientiﬁc can provide complete, integrated solutions with the Thermo Scientiﬁc OEM Controller, or using the manufacturers listed below.

• Applikon SUB-Controller

• Legacy Finesse PC Controller

• Emerson Delta V

• Allen-Bradley

• Siemens

Contact your local sales representative for more information.

Note: The S.U.F. 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.F. overview

1.4. BPC characteristics

1.4.1. S.U.F. BPC features

The microbial culture itself is contained inside the S.U.F. BPC (Figures 1.2 and 1.3). The chamber is manufactured from single-web, multi-layer 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, which is available from your sales representative). The BPC is presterilized using validated gamma irradiation processes.

1.4.2. Operating pressure

The S.U.F. 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.F. 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.F.

1.4.3. Working volume

Each S.U.F. is designed for a working volume range. The minimum working volume and the rated working volume are listed in the speciﬁcation tables provided in Chapter 4. The total volume listed includes the head space needed for proper aeration and gas management. Actual working volumes should not exceed the indicated rated working volumes. In addition, working volumes less than the minimums listed can result in damage to the BPC and S.U.F. hardware malfunction.

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Chapter 1 | S.U.F. overview

1.4.4. Draining and harvest

The S.U.F. is equipped with a bottom drain line that allows for liquid harvest, typically with a peristaltic pump. Connection of the bottom drain line can be accomplished by use of a tubing welder or the aseptic connection of quick connect or ﬁtting that is provided. Manipulation of the BPC as the last few liters of media are removed can minimize liquid hold-up within the S.U.F.

1.4.5. Aeration

Gas to liquid mass transfer in microbial culture fermentors is controlled by the solubility of the gas in the liquid, its distribution, and the temperature and pressure. Direct air sparging is a way of providing for the oxygen requirements of cell cultures. The standard S.U.F. BPC incorporates a unique rind-like ﬁlm sparger design that allows for optimal aeration of the culture process.

1.4.6. Aseptic connections

Multiple aseptic connection options exist for S.U.F. users. The standard BPCs include tubing welder sections, quick connects, CPC

™

AseptiQuik,

or Pall™ Kleenpak™ aseptic connections. The S.U.F. BPC is designed with various lengths and dimensions of thermoplastic tubing for the purpose of addition to and dispensing from the S.U.F. BPC. Refer to ordering information in Chapter 4 for custom end treatment options.

™

1.4.7. Sampling

The S.U.F. 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 an aseptic luer lock connector (SmartSite 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 by tubing welder.

™

) that allows

Detailed BPC drawings and standard conﬁguration tables speciﬁc to volume can be found in Chapter 4 of this guide.

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Chapter 1 | S.U.F. overview

1.4.8. BPC features

Figures 1.2 and 1.3 illustrate the components of standard 30 L and 300 L S.U.F. BPCs. See Table 1.2 on the following page for more information.

Figure 1.2. 30 L S.U.F. BPC.

Figure 1.3. 300 L S.U.F. BPC.

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Chapter 1 | S.U.F. overview

Table 1.2. BPC information for Figures 1.2 and 1.3.

1. Exhaust vent ﬁlter Single-use capsule ﬁlter for exhaust gas exchange (two types available)

2. Condenser bag (optional) Optional single-use condenser bag

Component Description

3. Foam sensor (optional)

4. Seal/bearing assembly

Built-in single-use sensor may be used with a controller to automatically trigger addition of an anti-foaming agent when excessive foam is present

Links with mixer motor and allows impeller to turn while retaining integrity of the S.U.F. BPC

5. Feed ports Feed port for addition of media and other liquids

6. Impeller

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

7. Ports with single-use sensors or standard Pall Kleenpak*

Ports with single-use sensors or standard Pall Kleenpak* connectors

connectors

8. Spare sterile connect port Spare port

9. Temperature RTD/ small volume sampling port

10. Gas sparge line(s)

For integration of temperature probe while retaining integrity of the S.U.F. BPC, and for needleless sampling or connection to sampling manifold

Drilled hole sparger integrated into the chamber and protected by gas ﬁlters and a check valve (check valve is provided on 30 L units only)

11. Drain port Used when draining the S.U.F.

*CPC AseptiQuik aseptic connectors are also available on S.U.F. BPCs.

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Chapter 1 | S.U.F. overview

1.5. Additional system components

1.5.1. Probe integration

The probe assembly is an innovative design to package user-supplied pH and DO probes for sterilization and to aseptically connect them to the S.U.F. BPC. The probe assembly (Figure 1.4) includes the following components:

1. Molded bellows cover

2. Threaded probe adapter

3. Pall Kleenpak connector (KPCHT series—high temperature)

4. Cable ties

Figure 1.4. Probe assembly.

Note: Figure 1.4 shows a Pall Kleenpak connector. CPC AseptiQuik connectors are also available for S.U.F. systems.

1.5.2. Accessories

To assist in the operation of the S.U.F., additional accessories are available (see accessories below).

Required

• Heavy-duty tubing clamps

Recommended

• Exhaust management system with condenser and vent ﬁlters.

Optional

• E-Box

• Sampling manifold with luer lock

• Temperature sample port (for RTD calibration/validation)

• Vent ﬁlter heater system—for users who require additional

protection for the exhaust vent ﬁlter on the standard S.U.F. BPC; includes ﬁlter heater, controller, and power cord.

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Chapter 1 | S.U.F. overview

• Load cells—Load cell packages include three load cells, summing

block, wiring, and an optional display screen with your choice of several data interfaces

• Cable management system

• Bottle management system

• Feed bag management hook

See Chapter 4 of this guide for accessory ordering information.

HyPerforma Single-Use Fermentor User’s Guide | 23Thermo Scientiﬁc

Chapter 2 | Installation and setup

Installation and setup

Chapter contents

2.1 Initial installation preparation

2.2 Installation

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Chapter 2 | Installation and setup

2.1. Initial installation preparation

The Single-Use Fermentor (S.U.F.) hardware will arrive crated. For unpacking instructions and detailed contents of the crate, please refer to the S.U.F. Packing and Unpacking Guide (DOC0052).

There will be detailed instructions for crating, uncrating, and assembly of 30 L and 300 L S.U.F. units. Be sure to follow the instructions provided and retain all packaging materials. After uncrating, contact your sales representative immediately if you ﬁnd that any damage has occurred in shipping.

2.1.1. Hardware shipment and setup

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

• Outer support container (platform, tank, and control panel)

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

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

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

shipped separately

2.2. Site preparation

2.2.1. Electrical connections for units with an electrical control panel

S.U.F. hardware with AC motors cannot be used on circuits equipped with GFCI circuit protection due to the potential for nuisance tripping. The electrical plug on the S.U.F. 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 serves to provide electrical ground prior to power connection. The supplied electrical receptacle should be hard-wired into the facility by a qualiﬁed electrical technician; for US 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 hard-wired directly into the facility. Note: The yellow plug and receptacle are for 120 VAC and the blue are for 240 VAC S.U.F.s.

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Chapter 2 | Installation and setup

2.2.2. Outer support container preparation

Each outer support container is shipped from the manufacturer and arrives with various safety mechanisms in place. Please follow the guidelines below to set up the S.U.F. upon arrival.

WARNING: Any procedures that require the electrical control panel

(E-Box) to be opened should be performed with the main electrical disconnect in the locked out position, and all power sources removed. For operator safety, secure the location of the S.U.F. unit by disabling the swivel casters before servicing.

2.2.3. Electrical preparation for systems with electrical control panels

1. Before beginning, refer to electrical schematics included with the

ETP, which is shipped on a USB drive.

2. Make sure the unit is disconnected from any power source, and

use standard lockout safety procedures.

3. Open the E-Box using a ﬂat-head screwdriver. Verify that the

three-way motor controller switch is in the middle position (Figure 2.1).

Motor controller breaker

Main breaker

Motor controller

Figure 2.1. Internal view of typical S.U.F. E-Box.

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Motor controller switch (3-way)

Chapter 2 | Installation and setup

4. Fermentor units are shipped with the main electrical breaker in the

off position. Turn the main breaker on.

5. Close the E-Box and lock the panel.

2.2.4. Load cell preparation

1. For S.U.F. units purchased with factory-installed load cells, the load

cells are shipped in the locked position (threaded up) for equipment protection. To unlock the load cells, remove and discard the delrin slip ring if it is present (Figure 2.2). Remove the tri-clamp. Loosen the lockout nut, using the small end of the supplied tool (Figure 2.3), until the nut is tight against the base or leg of the tank. Repeat this process for each load cell until all lockout nuts are disengaged from the lockout posts. Do not reinstall the tri-clamp.

Lockout nut

38.1 mm (1.5 in.) tri-clamp

Lockout post

Delrin slip ring

Drive shaft cap end

Figure 2.2. Load cells.

Load cell lockout end

Figure 2.3. Supplied wrench.

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Chapter 2 | Installation and setup

2. At this point, the S.U.F. hardware is ready to prepare for a cell

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

3. For systems with load cell display screens, refer to Appendix C for

information about how to calibrate the tools.

4. 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.

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

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

2.3. Installation

300 L S.U.F. systems are shipped with motors detached. Depending on doorway height, you may need to wait to install the motor until the S.U.F. is in place. See the speciﬁcations for the outer support container in Chapter 4 and the S.U.F. Packing and Unpacking Guide for more information.

All manual movements of mobile S.U.F. hardware should be over smooth surfaces, with the S.U.F. 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.F. Refer to the previous subsection of this guide for illustrations.

WARNING: Tipping hazard. The vessel should only be moved by pushing using the provided handles or at the mid-point of the vessel.

1. Verify that the facility electrical supplies are sufﬁcient to support the

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

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Chapter 2 | Installation and setup

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

run.

3. If monitoring the volume, the unit may be located on a scale if load

cells are not utilized. Other applications may measure all liquids going in and coming out.

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 cable/tubing

lengths are satisfactory.

WARNING: Electrical shock is possible.

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 S.U.F. should be connected to a dedicated 10 A circuit. Refer to hardware/electrical labels and schematics to ensure proper electrical voltage is connected to the S.U.F. The main power switch can now be turned on.

9. Connect the water inlet and outlet lines from the temperature

control unit (TCU) to the S.U.F. (Figure 2.4). The inlet is typically on the left side when you are facing the

connectors

. The inlet and

outlet ports on the S.U.F. water jacket are labeled.

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Chapter 2 | Installation and setup

Figure 2.4. Attaching a TCU hose to the water jacket port.

2. 3.1. Exhaust ﬁlter bracket setup

To install the exhaust ﬁlter bracket, clamp it onto the rim of the S.U.F. tank. Push the red handles down to lock it into place (Figure 2.5). The exhaust ﬁlter post will slide into the hole in the bracket. The center red handle locks the post in place. This may need to be adjusted during initial BPC loading. Note: The pinch clamp for 30 L systems must be installed onto the exhaust ﬁlter post before it is mounted to the tank. See section 2.3.4 for instructions.

Figure 2.5. Clamping the exhaust ﬁlter bracket to 30 L S.U.F. tank.

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Chapter 2 | Installation and setup

30 L condenser attached

to pole

2.3.2. Condenser unit setup (when present)

If your system includes an optional condenser, the assembled condenser will need to be installed. Condensers for 30 L units are clamped onto the exhaust ﬁlter pole (Figure 2.6). Condensers for 300 L units are bolted onto the exhaust ﬁlter bracket (Figure 2.7).

300 L

condenser attached to rim mount

Figure 2.6. Condenser installed on a 30 L system.

Figure 2.7. Condenser installed on a 300 L system.

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Chapter 2 | Installation and setup

2.3.3. TCU/chiller connection to condenser unit (optional)

Use the following steps to connect a TCU or chiller to the S.U.F. condenser unit, if present.

1. Connect inlet and outlet hoses to the TCU (Figure 2.10). You may need to attach the valves to the TCU, if necessary.

a. Use the clear supplied washers and tri-clamps to attach the ﬁtting

and then the hoses to both the inlet and the outlet port on the TCU (Figures 2.8 and 2.9).

Figure 2.8. Attaching line to a TCU unit. Figure 2.9. Attaching a clamp to a line.

Process outlet port on TCU

Process inlet port on TCU

Figure 2.10. TCU ports with lines connected for S.U.F. condenser.

2. Connect the remaining end(s) to inlet port(s) on the condenser.

closer

On 300 L systems the inlet ports are the ports (shown in Figure 2.11)—On 30 L systems the inlet port is the

to the tank

upper

port on the condenser (can be seen in Figure 2.6).

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Chapter 2 | Installation and setup

3. Connect the remaining end(s) to outlet port(s) on the condenser. On

300 L systems the outlet ports are the ports

farther from

(shown in Figure 2.11)—On 30 L systems the outlet port is the

to the tank

lower

port on the condenser (can be seen in Figure 2.6).

Note: The inlet and outlet hoses for 300 L systems include a Y-splitter to make the connections on the condenser plates.

Outlet ports on 300 L condenser

Figure 2.11. Attaching TCU lines to a condenser on a 300 L S.U.F. unit.

Inlet ports on 300 L condenser

4. Fill the TCU and prepare it for operation as speciﬁed in the user’s

guide provided with the TCU.

2.3.4. Exhaust ﬁlter pinch clamp setup (when present)

Some systems include an exhaust ﬁlter pinch clamp (Figure 2.12), which is used to temporarily keep exhaust from ﬂowing through redundant exhaust ﬁlters.

Pinching surface

Figure 2.12. Exhaust ﬁlter pinch clamp.

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Release button (

blue

light indicates unit is receiving power)

Locking pin

Chapter 2 | Installation and setup

Use the following steps to install the exhaust ﬁlter pinch clamp.

1. Mount the pinch clamp to the exhaust ﬁlter post (Figure 2.13) using the supplied C bracket. If the exhaust ﬁlter post is not already installed on the system, mount it onto the exhaust system bracket.

WARNING: Pinch hazard. Keep hands away from the pinching

surface. Close and use locking pin when pinch clamp is not in use.

Pinch clamp installed onto exhaust system post with C-bracket

Figure 2.13. Pinch clamp installed on a 300 L S.U.F.

2. Lift the center red handle of the exhaust bracket. Slide the exhaust ﬁlter holder post into the bracket and lock the center handle down.

3. Plug the wire attached to the pinch clamp unit into the controller. A blue light on the pinch clamp’s release button indicates that unit is receiving power and the unit is ready for manual initiation of the clamp.

Refer to the BPC loading instructions in section 3.2 for more information about how to use the pinch clamp with the BPC.

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Chapter 2 | Installation and setup

2.3.5. Optional cable management system setup

If your system includes the optional cable management system, the arm of the cable management system may be removed for shipping. Use an allen wrench to attach the arm as shown in Figure 2.14.

Refer to the BPC loading instructions in section 3.1 for more information on how to use the cable management system.

Figure 2.14. Attaching arm to cable management system.

2.3.6. Air line preparation

See Tables 3.2 and 3.3 (page 96) for recommended air ﬂow rates. The operating pressures at the level of the S.U.F. are of primary importance and these values must be adhered to. Please note that ﬂow rates in the graphics include both half and full volume applications.

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

should not be allowed to become tight during inﬂation or operation. Gas pressure in the BPC should not exceed 0.03 bar (0.5 psi) at any time. Conditions of over pressure may result in BPC damage or personal injury. For reference the BPC will appear to be tight at 0.007 bar (0.1 psi). The tubing after the inlet ﬁlter should be easy to compress with two ﬁngers. When testing, be sure NOT to pinch off the air inlet line completely.

2. 3 .7. Drilled hole sparger line(s)

Manually orient the sparge line vertically for maximum effectiveness.

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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 30 L units

3.3 BPC and drive shaft loading instructions for 300 L units

3.4 Exhaust system

3.5 Probe assembly

3.6 Microbial cell culture operating instructions

3.7 BPC sampling

3.8 Veriﬁcation procedures

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Chapter 3 | Operating information

3.1. General system operating information

3.1.1. BPC preparation and loading

Each outer support container is designed for a speciﬁc S.U.F. BPC. Conﬁrm that the correct volume BPC is being used for the corresponding volume outer support container. This section outlines the installation and setup of the different volume S.U.F. BPCs. Please follow these instructions in the order in which they are presented.

3.1. 2. BPC handling instructions

Use caution when using sharp objects to open outer polybags, in order to avoid causing damage to the BPC. Do not drag containers over corners or sharp objects. Do not lift the container by the corners or top seams. Carefully coil tubing on top of the BPC to prevent puncturing the container with cable ties or clamps. Use cushioning between tubing and container in storage and transport.

3.1.3. BPC operating information

Aseptic line connection

The most commonly recommended process for making connections to the tubing lines is with an aseptic tubing fuser. Note: Other connection options are available as a custom BPC assembly.

Following the recommended tubing welder operating instructions, successful connections can be obtained for ﬁlling, supplementing, sampling, or dispensing from the BPC as needed.

Working volume

Each S.U.F. is designed for a working volume range. The minimum working volume and the rated working volume are listed in the speciﬁcations tables in Chapter 4. The total volume listed includes the headspace needed for proper aeration and gas management.

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

Exhaust system

The exhaust vent ﬁlter(s) and condenser used on the 30 L and 300 L S.U.F.s include ZenPure ﬁlters. Condensate must be managed by use of the condenser system or vent ﬁlter heater(s).

™

polyethylene or Meissner™ PVDF

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Chapter 3 | Operating information

Sampling

The S.U.F. is equipped with a small volume sample port that is part of the BPC thermowell. This small diameter (1.59 x 4.76 mm (1/16 x 3/16 in.)) silicone dip tube of 152.4 mm (6 in.) length allows low void volume samples to be taken for cell viability and density, as well as analyte analysis. This dip tube is supplied with an aseptic 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 (3.18 x 6.35 mm (1/8 x 1/4 in.)) by tubing welder. For recommended systems for ﬂuid transfer, contact your sales representative.

Operating pressure

The BPC is not rated as a pressure vessel. All gas supplied to the fermentor controller must be regulated to a pressure not to exceed manufacturer’s recommendations, typically 1.034–4.137 bar (15–60 psi), depending on vessel scale and use. Gas pressure in the BPC headspace should not exceed 0.03 bar (0.5 psi) at any time.

WARNING: Conditions of overpressure may result in BPC damage or

personal injury. The maximum pressure in the tubing after the inlet ﬁlter should be about 0.344 bar (5 psi). The section of tubing after the inlet ﬁlter should be easy to compress with two ﬁngers.

The BPC is not operated as a closed system. It has both inlet and exhaust ﬁlters that are utilized to maintain a sterile 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 the failure of a pressure regulator on a gas feed, or when excessive foam within the S.U.F. creates conditions of vent blockage. The S.U.F. BPC is not rated as a pressure vessel. All gas supplied to the fermentor controller must be regulated to a pressure not to exceed manufacturer’s recommendations, typically 2.06–4.13 bar (30–60 psi). Gas pressure in the BPC headspace should not exceed 0.03 bar (0.5 psi) at any time.

• Demanding applications may warrant an optional exhaust vent

heater.

• If foaming is excessive in your cell culture process, use a foam

sensor.

• Single-use pressure transducers are available with the S.U.F. This

technology combined with high-level control systems common with industrial applications can regulate gas pressure within the conﬁnes of the S.U.F.

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Chapter 3 | Operating information

Extensive testing has not found an occurrence of overpressure sufﬁcient to create a containment breach. Development testing of the BPC system has shown that in conditions of excessive pressure the polymer container will fail at the upper regions of the chamber where it is unsupported by the outer support container, minimizing the likelihood of the loss of bulk liquid.

Agitation

The agitator should not be operated at volumes less than the stated minimum volume. Stirring speed may be adjusted by using the keypad interface on the controller. At low volumes, the chamber should be inﬂated to prevent the ﬁlm of the BPC from contacting the agitators.

Agitation control interface for units with electrical control panels

The agitation control interface utilizes an LED digital display to indicate stirring speed in units of revolution 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 up and down arrows on the agitation keypad adjust the stirring speed. Due to the auto-restart capabilities of the S.U.F., the green start button on the keypad has been disabled; however, the red stop button on the keypad is active. If the red stop button has been used to stop the motor, the controller can be reset and agitation restarted by using the main motor toggle switch on the left side of the E-Box.

Dispense and harvest

The S.U.F. 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 aseptic connection of 6.35–9.53 mm (1/4–3/8 in.) quick connect that is provided. Manipulation of the S.U.F. BPC as the last few liters of media drain will minimize liquid hold-up within the S.U.F.

3.1.4. Hardware operating information

Temperature

Temperature setpoints can be adjusted via the temperature control unit (TCU). These setpoints should be between 5 and 55°C. The process temperature is measured by means of a supplied resistance temperature detector (RTD) (pt-100) that is inserted into the thermowell of the S.U.F. BPC. The temperature measured by the RTD is not displayed, but is passed through the 16-pin cable as a raw PT100 signal.

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Chapter 3 | Operating information

Heating performance

Heating times for the S.U.F. system vary based upon liquid volume and temperature, ambient or heating liquid temperature, sparge rate, and mixing rate. Users should adjust process liquid staging and seeding strategies to the unique aspects of the S.U.F. Process controllers and heaters are designed to provide optimum heat transfer and minimize heat up times, while maintaining the material integrity of the polymer ﬁlm construction of the BPC (Table 3.1).

Do not operate the heater if the BPC is not at minimum liquid, 5:1 (20%) volume or greater. Care must be taken not to melt or damage the container or other components. Heaters should not be used to warm liquid above 45°C. The main power disconnect should be in the off position unless the S.U.F. hardware system is in active use.

Table 3.1. Heating times for S.U.F. systems (Ambient temperature of 25°C).

System

Liquid batch volume

(half/full)

Initial liquid

Liquid target

Time (half/

full)

30 L 15 /30 L 5°C 37°C 1 hr/1.16 hr

300 L 150 /300 L 5°C 37°C 1.3 hr/1.8 hr

General temperature mapping has been performed for S.U.F. systems by tracking thermal proﬁles within the liquid conﬁnes of the fermentor. Testing conditions were analyzed using chilled media and low agitation rates. Gradients within the liquid did not exceed 0.5°C during heat-up. Testing has also shown that temperature measurements in the S.U.F. when using the standard silicone thermowell with 3.18 mm (1/8 in.) diameter temperature probes properly represent fermentor content temperatures. Users desiring exact temperature calibrations can order a S.U.F. temperature/sample port. Using this port will allow users to simulate the temperatures seen by the RTD when used with the BPC.

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Chapter 3 | Operating information

Protective earth grounding

Protective earth grounding for the S.U.F. 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 tank, and must be tied to the controller before operation.

Resettable breakers

Electrical components of the S.U.F. are equipped with circuit protection. The variable frequency drive (VFD) used to power the mixer motor is protected by the use of 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. The breakers can be reset once the fault condition is resolved.

Breaker notes:

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

• A tripped breaker will be in the mid-position.

• “Off” is in the fully down position.

• To reset a tripped breaker, it must ﬁrst be moved from the tripped

or mid position to the “off” or fully down position before moving it the fully up or “on” position.

Scales and weighing systems

Monitoring liquid volume within the S.U.F. during operation can be critical in fermentation applications that involve nutrient media feeds. This can also be a useful method for increasing the scalability of the S.U.F. 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 fermentor is increased to rated working volume during the process run.

A load cell kit for weight/volume measurement is available as an option. Load cell kits 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. For more information, refer to the Speciﬁcations and parts information in Chapter 4 of this guide. Refer to section 2.1 for information about unlocking the load cells, and Appendix B for load cell display calibration instructions. Note: Ensure that load cells are

locked down before moving the S.U.F. unit.

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Use the following steps to lock the load cells before transporting a S.U.F. (refer to Figures 2.3 and 2.4 in section 2.2.4 (page 27).

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 you hand-tighten the nut 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 over-tighten the nut.

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

the ﬂanges.

4. Repeat these steps each load cell attached to the tank.

External data logging and control

Optional 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.F., ﬂoor scales can be used to measure the ﬂuid content within the S.U.F. This is accomplished by rolling the S.U.F. onto the scale platform and leveling the S.U.F. skid once it is in position.

S.U.F. hardware systems are designed to allow advanced users to control all aspects of the operation of the fermentor. Contact technical support for Thermo Scientiﬁc products for general integration guidance.

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Chapter 3 | Operating information

3.2. BPC and drive shaft loading for 30 L units

3.2.1. Initial installation steps for 30 L units

Each outer support container is designed for a speciﬁc S.U.F. BPC. Conﬁrm that the correct volume BPC is being used for the corresponding volume outer support container. The following section outlines the installation and setup of the BPC.

Before beginning to load the BPC, verify that the S.U.F. hardware is stationary, with the casters locked into place.

1. Remove the irradiated BPC from the protective double polybags.

Open the S.U.F. door and load the BPC into the stainless steel outer support container (Figures 3.1 and 3.2).

Figure 3.1. Opening the S.U.F. door latch.

Figure 3.2. Interior of the outer support container.

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2. Orient the BPC with the bearing port up and toward the motor

drive, with the connector probe ports facing the bottom access cutout.

3. Remove the cap on the BPC bearing port, and place the bearing

port into the bearing port receiver (Figure 3.3). Close the door and the clamp.

Figure 3.3. Inserting the bearing port.

4. Route the bottom ports through the opening in the bottom of the

outer support container (Figure 3.4).

Figure 3.4. Routing the bottom ports through the bottom of the outer support container.

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5. Route the sparge line and bottom drain line through the bottom

access opening (Figure 3.5).

Figure 3.5. Routing the sparge and bottom drain lines.

6. Open the door of the condenser unit, if present, and feed the

condenser portion of the bag through the condenser.

7. Insert the vent ﬁlter(s) into the ﬁlter bracket (Figure 3.6).

Figure 3.6. Inserting a vent ﬁlter into the ﬁlter bracket.

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8. If present, straighten and smooth the condenser bag in the

condenser unit and close and latch the door of the condenser (Figure 3.7). Ensure that the condenser BPC is not too tight or too loose in the condenser hardware unit.

Figure 3.7. Loading of an optional condenser bag.

9. Secure ﬁlter heater(s) on the ﬁlter(s) (Figure 3.8). 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 or the E-Box.

Filter notes:

• The vent ﬁlter heater controller is preset to 55°C.

• 30 L BPCs may have one or two exhaust ﬁlters.

Figure 3.8. Installing an exhaust ﬁlter heater.

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10. If a cable management system is available, attach the lines to the

appropriate inlet ports (Figures 3.9, 3.10, and 3.11).

Figure 3.10. Incoming line connection.

Figure 3.9. Line setup on the cable management system.

Figure 3.11. Incoming line connection to a direct sparger ﬁlter.

11. Connect incoming gas feed line to the sparger ﬁlter. Inﬂate the BPC

with air through the sparger. Inﬂation time is approximately one to ﬁve minutes. Do not exceed 25 slpm or 0.034 bar (0.5 psi) internal pressure for 30 L BPCs. As the BPC inﬂates, ensure it is properly oriented in the support container (port alignment, drain, and sparger).

WARNING: DO NOT EXCEED 0.034 bar (0.5 psi) within the BPC or the

BPC could fail. The BPC will appear to be tight at 0.007 bar (0.1 psi). The pressure in the tubing after the inlet ﬁlter should be about 0.34 bar (5 psi) and should be easy to compress with two ﬁngers.

12. As the container ﬁlls with air, manipulate the container to align the

sparge line in the slot.

Note: While a check valve is provided for the sparge line on 30 L units, it is not uncommon for some ﬂuid to bypass check valves during typical use. Elevate the ﬁlter above the liquid level in the S.U.F. and secure it to prevent exposing the ﬁlter to liquid.

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Chapter 3 | Operating information

13. Use the bottom cutouts located at the base of the support

container as a reference to align the hanging tabs on the BPC.

14. Attach the two hanging tabs on the bottom of the BPC to the tab

holders (pins) on the S.U.F. (Figure 3.12).

Figure 3.12. Aligning a hanging tab with the pin.

15. Align the row of connector ports and probes, and then close the

S.U.F. door (Figure 3.13). Note: Verify that all port clamps are closed and located as close as possible to the body of the BPC.

Figure 3.13. Aligning the connector ports and probes.

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16. To allow for drive shaft insertion, ﬁll the BPC with air for

approximately 10–20 minutes (times will vary based on ﬂow rate, inlet pressure, and container volume).

WARNING: To prevent damaging the BPC, the BPC must be partially

inﬂated before you begin to insert the drive shaft. You must close and latch the access door of the S.U.F. before fully inﬂating the BPC.

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

sample line of the BPC. This grounds the media inside the BPC and helps to eliminate electrostatic charge (Figure 3.14).

Media ground clip connection

Figure 3.14. Media ground clip connection.

Use of exhaust ﬁlter pinch clamp (when present)

If your system includes a pinch clamp, follow the instructions below to close off redundant exhaust ﬁlters on the BPC.

1. Install the exhaust ﬁlter pinch clamp on the vent ﬁlter post.

2. Check to ensure that the blue light on the release button is

illuminated, which indicates that power to the unit is enabled. The clamp will be manually closed to a 0.159 cm (1/16 in.) clearance. The clamp will then automatically close the remaining gap to shut off the exhaust ﬁlter(s).

3. Remove the locking pin and open the clamp.

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4. Position the alignment cutout on the exhaust ﬁlter BPC, over the pin

on the left side of the clamping surface for 30 L units (Figure 3.15). Note: On 300 L units, this pin is at the top of pinch clamps.

Figure 3.15. Positioning the BPC cutout over the alignment pin in a pinch clamp.

WARNING: Pinch hazard. When using the pinch clamp, keep hands

away from the pinching surface.

5. Close the front of the clamp (Figure 3.16) and replace the locking

pin (Figure 3.17).

Figure 3.16. Closing the front of the clamp.

Figure 3.17. Replacing the locking pin.

6. Verify that the blue light is on.

7. Push the front of the clamp toward the pinching surface until it

almost touches the BPC (Figure 3.18). The clamp will automatically close completely.

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Figure 3.18. Pushing the pinch clamp closed.

8. When using the system, if the ﬁrst set of ﬁlters become fouled, the

pinch clamp can be manually opened to enable the backup ﬁlter. This should only be done when the pressure in the BPC is 0.034 (0.5 psi) or 0.041 bar (0.6 psi), and no foam is detected in the headspace. To manually open the clamp, push the illuminated blue button.

9. If the pinch clamp has been integrated into a controller, it can also

be released using a button on the controller’s touchscreen.

3.2.2. Drive shaft insertion for 30 L units

WARNING: To prevent damage to the BPC, the BPC must be partially

inﬂated before beginning to insert the drive shaft.

1. Remove the latch pin from the motor assembly (Figure 3.19).

Figure 3.19. Removing the latch pin from the motor assembly.

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2. Unscrew the threaded cap covering the pass-through of the motor.

3. Use the following steps to insert the drive shaft through the

pass-through of the motor (Figures 3.20 and 3.21).

• 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 prevent stretching of the impeller tubing.

• When approximately 30 cm (12 in.) of shaft remains, twist back and forth slightly to engage the impeller.

• When approximately 25–50 mm (1–2 in.) of shaft remains, twist back and forth slightly to engage the bearing assembly.

• When approximately 6 mm (0.25 in.) of shaft remains, twist to align the motor drive keyway with one of the four outer slots on the drive shaft head.

Figure 3.20. Using two hands to load the drive shaft.

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Figure 3.21. Aligning the motor drive keyway.

Chapter 3 | Operating information

4. Use the following steps to directly couple the drive shaft to the motor drive (Figures 3.22–3.24).

• Place the threaded cap on the hollow pass-through and hand-tighten in a clockwise direction (Figure 3.22).

• Tighten the cap by placing the spanner wrench on the hollow pass-through and tighten the cap using the supplied torque wrench (Figure 3.23). Note: The torque wrench is a standard 10 mm (3/8 in.) square drive, and is calibrated at the factory at

16.9 Nm (150 in-lb.).

• Verify that the wrenches have been removed from the system and placed in storage holders.

• Close the safety access cover and insert the latch pin (Figure 3.24).

Figure 3.22. Replacing the cap.

Figure 3.23. Tightening the cap. Figure 3.24. Replacing and latching

the cover.

5. The air supply may be turned off once the drive shaft has been inserted.

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3.2.3. Final installation steps for 30 L units

1. Position and close a bar clamp on the bottom drain line as close as possible to the BPC port (Figure 3.25).

Figure 3.25. Closing bar clamp on drain line.

2. Remove the plastic insert (if present) located in the thermowell.

3. Use the following steps to insert the RTD or selected temperature sensor into the thermowell (Figures 3.26 and 3.27).

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

• Insert the sensor until the base of the probe meets the mouth of the thermowell (Figure 3.26).

• Secure by twisting the luer lock collar, if provided. The thermowell will stretch slightly when the RTD is seated (Figure 3.27).

Figure 3.26. Inserting the RTD into the thermowell.

Figure 3.27. Securing the RTD by twisting the luer lock collar.

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4. Optional: Connect the pressure sensor to the monitoring device.

WARNING: Do not over-inﬂate the BPC, as it could rupture. An

automatic high-pressure gas shutoff device is recommended.

5. Refer to section 3.5.3 for probe insertion instructions.

6. Plug the optional foam sensor wire into the port in the top of the BPC (Figure 3.28), and attach the ground clip to the stainless steel insert in the RTD/sampling port (Figure 3.29).

Figure 3.28. Foam sensor wire plugged in.

Figure 3.29. Vessel to media ground clip.

RTD/sampling port

BPC ground clip

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3.3. BPC and drive shaft loading for 300 L units

3.3.1. Initial installation steps for 300 L units

Before beginning to load the BPC, verify that the S.U.F. hardware is stationary with casters locked into place. BPC loading may require operators to step inside the fermentor, and the unit must be stationary for the safety of both the operator and the equipment. For ease in BPC loading, two operators may be required. For drive shaft insertion and exhaust ﬁlter positioning, a ladder or other means of elevation is required.

1. Open the door of the S.U.F. outer support container (Figure 3.30).

Figure 3.30. Opening the fermentor door.

2. Release (pull down) the latch that locks the motor in place, and slide the motor away from the center of the tank.

3. Remove the irradiated BPC from its protective packaging and double polybags (Figure 3.31). Do not remove 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.

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Figure 3.31. BPC in protective polybags.

4. Reach into the outer support container with the front face (bearing port side) of the BPC oriented towards the motor (Figure 3.32).

Figure 3.32. Bearing port orientation.

Note: These steps may also be performed from the top of the unit. Be sure to use caution when using a ladder. Maintain three points of contact, and ensure that the steps are clean.

5. Place the top line sets (still in polybags) over the top edge of the tank (Figure 3.33). This will keep the container from being restricted during the air inﬂation step.

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Figure 3.33. Preparing to place bagged lineset over edge of container.

6. From inside the tank or from an elevated position, slide the exhaust port into the exhaust port bracket at the top of the outer support container (Figure 3.34). Slide the black lever to lock the exhaust port into the bracket (Figure 3.35 and 3.36). It should click into place.

Figure 3.34. Inserting the condenser into the exhaust port in a holder at the top of the tank.

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Exhaust port latch (closed)

Figure 3.35. Exhaust port sliding latch (removed from the tank).

Slide latch to lock exhaust port in place

Figure 3.36. Locking the exhaust port.

7. Remove the cap from the bearing port in the BPC and load the bearing port into the receiver (Figure 3.37). The motor may be locked into position, or unlocked to facilitate bearing port insertion, if preferred. Close the bearing port door and latch it (Figure 3.38).

Figure 3.37. Bearing port in the receiver.

Figure 3.38. Latching the bearing port.

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8. If the motor was unlocked to facilitate insertion of the bearing port, lock the motor back into position.

WARNING: Failing to lock the motor into position may cause harm to

personnel or damage the BPC.

9. If your unit has one, open the door of the condenser. Feed the condenser bag through the condenser, and attach the exhaust ﬁlter bags into the ﬁlter bracket (Figure 3.39). Then close and lock the condenser door (Figure 3.40) and adjust the height of the ﬁlter bracket so there are no wrinkles or slack in the exhaust system.

Figure 3.39. Exhaust ﬁlter and condenser setup.

Figure 3.40. Closing the condenser door.

10. Remove the white plastic guide plate from the bottom of the outer support container (Figure 3.41) and guide the sparge inlet line, ﬁlter, and the drain line through the bottom cutout in the outer support container. Replace the plastic guide plate (Figure 3.42).

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Drain line cutout

Sparge line cutouts

Figure 3.42. Replacing the guide plate.Figure 3.41. Removing the guide plate.

11. Extend the tubing through the bottom of the outer support container. Ensure that the tubing isn’t pinched or kinked under the container (Figure 3.43).

Figure 3.43. Sparge and drain line extension.

12. Pull the temperature/sampling port through the large opening in the front of the outer support container.

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Optional: If you are using the optional exhaust ﬁlter pinch clamp to control airﬂow to redundant ﬁlters, follow the steps listed in section 3.2.1. The same pinch clamp is used for both S.U.F. sizes. However, the clamp is oriented vertically on the 300 L system (Figure 3.44), and horizontally on the 30 L system.

13. Connect a pressure sensor to the monitor (or the third-party

Figure 3.44. Pinch clamp installed on a 300 L BPC.

controller, if used). After the display has stabilized, allow it to warm up for 30 minutes, and then tare the monitor. Verify that the monitor reads zero. Note: Filter input air pressure can be up to about

3.103 bar (60 psi) for the 300 L S.U.F. The maximum pressure in the tubing after the inlet ﬁlter should be less than 0.334 bar (3 psi), and that section of tubing should be easy to compress with two ﬁngers.

WARNING: DO NOT EXCEED 0.34 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. When inﬂating the BPC, ensure that the tubing isn’t pinched or kinked under the outer support container.

Note: Filling the BPC with air takes approximately 15–20 minutes before drive shaft insertion can begin. Times will vary based upon ﬂow rate and inlet pressure.

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14. Remove the bubble wrap from the inlet ﬁlter and sparge line (Figure 3.45), connect to gas supply, and partially inﬂate the BPC. This will allow for proper insertion of the drive shaft and aid in aligning the BPC in the outer support container.

Figure 3.45. Sparge line and ﬁlter.

15. As the BPC ﬁlls with gas, attach the hanging tabs on the BPC to the hooks (pins) on the S.U.F. hardware at the top and bottom of the unit (Figure 3.46).

Figure 3.46. Attaching hanging tabs.

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3.3.2. Drive shaft insertion for 300 L units

The motor and mixing assembly in Figure 3.47 serves as a reference for drive shaft assembly and insertion. The drive shaft is constructed using three pieces, which must be assembled and inserted piece by piece. Operators should be elevated (i.e. ladder) to effectively assemble and insert the drive shaft.

WARNING: Review ceiling height requirements in Table 4.2 in

section 4.2 before trying to insert the drive shaft. Use caution when on a ladder or elevated platform. Ensure that it is stable, the steps are clean, and maintain three points of contact.

Drive shaft head

Drive shaft cap

Motor drive keyway

Hollow passthrough

Safety cover

Figure 3.47. Motor and mixing assembly.

Latch pin

Motor

Drive shaft

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1. Prepare the hollow pass-through by ﬁrst removing the latch pin on the safety cover (Figure 3.48), opening the safety cover, and removing (by turning counter-clockwise) the threaded drive shaft cap of the mixing assembly (Figures 3.49 and 3.50).

Figure 3.48. Removing the latch pin. Figure 3.49. Opening the drive shaft

cap.

Figure 3.50. Removing the drive shaft cap.

2. Verify that the three sections of the drive shaft are located in the drive shaft holders on the side of the outer support container. The sections will be referred to as the upper (section with the drive shaft head), middle (section with external threads on each end), and lower (section with the hexagonal end). Lubricate the threaded ends with a light coat of food grade anti-seize with each use.

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3. First, insert the lower section through the hollow pass-through of the motor drive. Slide the latch pin from the motor assembly into the shaft to prevent it from falling into the tube (Figure 3.51). Assemble the middle and lower sections of the drive shaft by joining the segments with a twisting motion, fastening the two sections together (Figure 3.52).

Locate one wrench on the ﬂat area in the middle drive shaft section, another wrench on the lower section, and tighten the connection using a counter-clockwise rotation (Figures 3.53 and 3.54). The shafts are left-threaded (reverse-threaded) to avoid loosening during operation. Once the sections are secure, return the wrenches to the tool holder. CAUTION: Do not over-tighten; a snug ﬁt is sufﬁcient. Remove the latch pin.

Figure 3.51. Inserting the lower section and securing it with a latch pin.

Figure 3.53. Tightening the drive shaft connections.

Figure 3.52. Connecting the middle section of the drive shaft.

Figure 3.54. Connecting the upper drive shaft segment.

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

5. Using two hands, carefully guide the completed drive shaft into the BPC using a slight back and forth twisting motion (Figure 3.55). 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 when the end of the drive shaft begins to couple with the impeller.

• When 50.8 cm (20 in.) of shaft remains, twist slightly to engage the impeller.

• When 25–50 mm (1–2 in.) of shaft remains, twist slightly to engage the bearing assembly.

• When 6–12 mm (0.25–0.5 in.) of shaft remains, twist to align the motor drive keyway with one of the four outer slots on the drive shaft head.

Figure 3.55. Aligning the motor drive keyway with the drive shaft head slots.

6. Directly couple the drive shaft to the motor by placing the threaded cap back on the hollow pass-through and tighten (Figure 3.56).

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Figure 3.56. Replacing the drive shaft cap.

7. Tighten the cap by placing a spanner wrench counterclockwise on the hollow pass-through. Tighten using the supplied torque wrench (Figure 3.57). Note: The torque wrench is a standard 10 mm (3/8 in.) square drive, and it is calibrated at the factory at 16.9 Nm (150 in-lb.).

Figure 3.57. Tightening the cap with wrenches.

8. Remove the wrenches from the system and return them to the storage holders.

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

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3.3.3. Final installation steps for 300 L units

1. Fully extend the ports through the front cutout of the outer support container, and attach the probe clips (Figure 3.58).

Figure 3.58. Attaching a probe clip.

2. Remove the drain line set from the polybag (Figure 3.59), position the line clamp as close as possible to the BPC port, and close the clamp (Figure 3.60). Use a cable tie around the clamp to ensure that it cannot be accidentally opened.

Figure 3.59. Removing the drain line set from a polybag.

Figure 3.60. Placing a clamp on the drain line.

3. If present, remove the plastic insert located in the thermowell.

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Chapter 3 | Operating information

4. Use the following steps to insert the RTD or selected temperature sensor into the thermowell (Figure 3.61).

• Place a small amount of glycerol (0.5 mL) in the well to aid in heat transfer.

• Insert the sensor 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 3.61. Inserting a sensor into the thermowell.

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

6. If your BPC is equipped with a foam sensor, connect the foam sensor wire to the port at the top of the BPC (Figure 3.62).

Figure 3.62. Foam sensor wire connected in port.

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7. Attach the ground clip to the stainless steel insert in the RTD/ sampling port (Figure 3.63).

RTD/sampling port

BPC ground clip

Figure 3.63. Foam sensor ground clip on the RTD.

8. If your system includes a cable management system, refer to the instructions for setting up the cable management system in the optional cable management system setup in section 2.3.5 of this guide.

Note: Since there is no check valve on the inlet gas line for the 300 L S.U.F., position inlet ﬁlters well above the liquid level to prevent fouling of the ﬁlters.

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Chapter 3 | Operating information

3.4. Exhaust system

3.4.1. Exhaust system functional overview

The condenser is intended to be used as an accessory to the S.U.F. in conjunction with vent ﬁlter heaters, which are necessary with or without the condenser. The condenser’s purpose is to prevent liquids and solids from condensing and collecting inside of the vent ﬁlters of the S.U.F.

The condenser cools the exhaust gases leaving the S.U.F. chamber, condensing the moisture out of the saturated gases coming from the S.U.F. The liquid condensate that is stripped from the exhaust gases is then returned to the bioprocessing container chamber, creating a sterile loop and signiﬁcantly reducing liquid loss due to evaporation. The condenser plate is chilled by a closed bath recirculating chiller. Figure 3.64 is a functional diagram of the exhaust system on a 300 L S.U.F.

Exhaust vent ﬁlters

Vent ﬁlter heaters

Condenser container bag

Condenser plate assembly

Condenser bleed plug

Dual headed peristaltic pump

Thermal Control Unit (TCU)

Filter bracket assembly

Condenser post assembly

Exhaust port from BPC chamber

Condenser return line back to S.U.F.

Condenser bag liquid drain ports

Cart assembly

Figure 3.64. Complete exhaust system on a 300 L S.U.F.

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Chapter 3 | Operating information

3.4.2. When to use the condenser

Large 254 mm (10 in.) hydrophobic polyethylene vent ﬁlters with a nominal 0.2 micron pore size increase the available surface area for off-gassing. In addition, the standard 30–300 L S.U.F. is designed for use with an optional condenser. This allows the S.U.F. to improve exhaust vent protection and reliability because it strips condensate and atomized materials that may be present from the off-gas stream of the S.U.F. This system has been shown to signiﬁcantly reduce the “fouling” load on the vent ﬁlters that may also increase operating back pressure as the cell culture run progresses. See the S.U.F. Validation Guide for speciﬁc details.

Some end users may prefer to omit the condenser under the expectation that this will allow for a more uniform installation or will reduce system complexity and cost. The use of exhaust vent heaters and 254 mm (10 in.) ﬁlters will provide some impressive ﬂow capacity over short periods. However, the high sparge rates required may eventually create conditions of increased operating back pressure, usually due in part to blocking of the ﬁlter media. Depending on the application, the user may use both ﬁlters in parallel or initiate the run with a single ﬁlter, temporarily clamping off the line to the other ﬁlter, and reserve it for use as redundant back-up.

Various vent ﬁlter conﬁgurations are available on the S.U.F., depending on the process scale and intended application. In all cases, using a vent ﬁlter heater will reduce the chance of condensate blocking the ﬁlter. 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. S.U.F. BPCs may be equipped with foam sensors that monitor foam in the head space and can be used to automatically trigger the addition of an anti-foaming agent. A vent ﬁlter heater has very little tolerance for handling the presence of foam in the exhaust stream. A small feed of antifoaming agent added directly to the liquid surface of the head space typically provides excellent foam control.

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Chapter 3 | Operating information

3.4.3. Condensers and reserve vent ﬁlters

All systems, especially large systems, beneﬁt from the use of a condenser. The condenser increases system reliability at high ﬂow rates beyond 1 vvm (or 30 splm for 30 L vessels, and 300 slpm for 300 L vessels), and warrants strong consideration when performing batch runs beyond one day at 1 vvm (or three days at lower vvms). Results will vary. However, it is strongly recommended that end users select a vent ﬁlter conﬁguration providing reserve capacity where possible. For example, on 30 L units, dual vent conﬁgurations can be used independently, with the second ﬁlter serving as a backup (providing a quick reserve in case issues arise in-process). For 300 L S.U.F.s, a single vent ﬁlter may be used at 300 slpm (1 vvm), but a second ﬁlter may be added to serve as a backup. At 600 slpm (2 vvm), two ﬁlters are required, but two additional ﬁlters may be added as backups.

3.4.4. Exhaust system setup

1. Remove and assemble the vent ﬁlter holder and condenser from the crate or shipping box. Assembly instructions are included in the S.U.F. Packing and Unpacking Instructions document.

Note: For safety, two people should assemble the 300 L condenser onto the S.U.F.

2. If your system includes a condenser, connect the TCU hoses to the ports on the condenser unit.

3. Remove the reservoir cap of the TCU and add the appropriate type and volume of ﬂuid, as speciﬁed in the TCU user’s guide.

4. Verify that the peristaltic pump, if used, and TCU power cords are connected to power.

5. Turn on power to the TCU. This will allow the TCU to prime.

6. Use the processes in the TCU user’s guide to set the pressure, between 1.37–2.06 bar (20–30 psi). Normally, this is controlled by a valve on the back of the unit, and displayed on an LCD display on the front of the unit.

7. Add ﬂuid to the reservoir in the TCU as needed, in order to maintain the minimum level.

8. Purge the condenser plate by loosening the bleed plug on top of the plate. This is accessed using an open-end wrench. Loosen the plug only enough to allow trapped air to escape, then re-tighten.

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Chapter 3 | Operating information

9. Check the indicator or window on the front of the TCU to ensure that the reservoir is still at least at the minimum level.

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

3.4.5. Loading the condenser BPC

1. Remove the BPC with the condenser container (if ordered) and vent ﬁlters from the polybag packaging, being careful not to puncture the container.

2. For 30 L units, load the BPC into the bearing hub as described in section 3.2 of this guide. For 300 L units, it is best not to load the BPC into the bearing hub until the condenser has been loaded, but place the BPC into the main S.U.F. tank.

3. For 300 L units with condensers, ﬁrst insert the exhaust port of the BPC into the exhaust port holder, which is attached to the bracket used to clamp the exhaust system onto of the outer support container (hardware). It may be helpful to slide the motor back out of the way for this step.

4. Open the door of the condenser unit, if present (Figure 3.65). Position the rectangular container inside the condenser unit, being careful not to pull the container out of shape. The vent ﬁlter can hang freely. Leave the condenser door open.

Figure 3.65. Opening the condenser door (300 L S.U.F. unit).

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5. Place the top(s) of the ﬁlter(s) into the ﬁlter bracket(s) (Figure 3.66). Adjust the height of the ﬁlter bracket, as needed. Make sure that the position of the ﬁlter is not pulling the BPC condenser too tight, pinching off the air ﬂow. It should also not be too loose, causing it to kink.

Figure 3.66. Positioning the condenser BPC and vent ﬁlter in bracket.

6. For units with condensers, ensure that there are no kinks or wrinkles in the container. Then, close and latch the condenser door (Figure 3.67).

Figure 3.67. Closing and latching the condenser door.

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Chapter 3 | Operating information

7. Place the vent ﬁlter heater(s) around the ﬁlter(s), looping the top strap of the heater around the metal bar on the ﬁlter bracket, and using the snap to secure them in place (Figures 3.68 and 3.69).

Figure 3.68. Snapping a vent ﬁlter heater onto the ﬁlter.

Figure 3.69. Using the snaps to secure the vent ﬁlter heater.

8. For 300 L units with condensers, load the tubing into the peristaltic pump, ensuring that there is sufﬁcient slack at each end of the pump tubing. Align the tubing in the pump channel, and close the pump ramp (Figure 3.70). Verify that the ﬂow direction returns condensate to the vessel. Consult the pump user’s guide for more information about operating the peristaltic pump.

30 L note: For 30 L units, a peristaltic pump is not needed, as condensate drips back into the main BPC chamber.

Tubing note: Ensure that the tubing size is correct for the pump

™

head. PharMed

tubing is recommended for the condensate line.

Standard tubing is silicon, which should be sufﬁcient for 24 hours.

Figure 3.70. Loading pump tubing.

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Chapter 3 | Operating information

9. Verify that both the pump and TCU or chiller are enabled and running at the proper settings. We recommend a pump setting of 12–30 rpm, and a TCU/chiller setting of 5°C. Condensate ﬂow should be veriﬁed at the maximum gas ﬂow settings.

10. After setting up the 300 L unit, verify that the pump union is loose on both ends of the pump, and running smoothly in the peristaltic rollers.

Speciﬁc performance questions can be answered by reviewing the condenser section of the S.U.F. Validation Guide, or by contacting technical support.

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Chapter 3 | Operating information

3.5. Probe assembly

3.5.1. Preparation and sterilization

1. Select the appropriate probe. Verify the presence of a Teﬂon™ support ring and O-ring on the probe and visually inspect them for damage.

2. Perform any required probe maintenance, and calibrate the pH probe (see the probe calibration topic in section 3.5.4 of this user’s guide).

3. Insert the probe into the probe assembly through the threaded adap to r.

4. Verify that the probe tip is not touching the membrane of the aseptic connector (more than 6.35 mm (1/4 in.) gap) before threading it into the probe adaptor.

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

6. Place the probe assembly with probe into the autoclave tray for probe kits (Figure 3.71).

Probe assembly

Handle

Autoclave tray for probe kits

Figure 3.71. Probe and autoclave tray.

7. Autoclave the probe assembly using a validated sterilization cycle, approximately 30 minutes at 122°C. 30-minute sterilization cycles are generally sufﬁcient. Options of wet or dry cycle parameters can be used. Slow exhaust cycles are preferred, as this minimizes stress on the probes during the temperature and pressure changes of autoclaving.

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Chapter 3 | Operating information

8. Allow sufﬁcient time for probe assemblies to cool completely before connecting to the BPC for probe insertion.

9. When stored properly, the autoclaved probe assemblies can be stored dry for short periods of time (less than 24 hours) without loss of sensor longevity, performance, or sterility.

3.5.2. Making CPC AseptiQuik connections

CPC AseptiQuik G genderless connector components

™

Figures 3.72 and 3.73 below illustrate the components of CPC

™

AseptiQuik

G genderless connectors. Connectors with white protective cover pull tabs may be autoclaved. Generally, connectors with blue pull tabs are gamma irradiated, not autoclaved. Visit the Colder Products Company website at http://cpcworldwide.com for more information.

For instructions on making an aseptic connection, see the next section.

Paper membrane

Hinge

Protective cover pull tabs

Figure 3.72. CPC AseptiQuik G connector (closed).

CPC AseptiQuik connection instructions

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

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

located on the BPC (Figures 3.74 and 3.75).

Figure 3.73. CPC AseptiQuik G connector (open).

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Chapter 3 | Operating information

Figure 3.74. Pulling the tear strip.

2. Unsnap and ﬂip open the protective cover pull tabs on both

connectors (Figures 3.76 and 3.77).

Figure 3.76. Opening the protective cover pull tab on the port.

Figure 3.75. Removing the plastic covering from the connector.

Figure 3.77. Opening the protective cover pull tab on the line set.

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Chapter 3 | Operating information

3. Align the two connectors and push them together.

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

(Figure 3.78).

Figure 3.78. Squeezing connectors together.

5. Grab the joined pull tabs and pull upward to remove the paper

membranes from the connectors (Figure 3.79). The pull tabs will also be removed.

Figure 3.79. Pulling tabs to remove the paper membranes.

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Chapter 3 | Operating information

3.5.3. Probe insertion

Before beginning probe insertion, please become familiar with the CPC AseptiQuik connector procedure outlined in section 3.5.2. If you are using Kleenpak connectors instead, refer to section 3.5.5.

1. Attach probe clips onto the outer support container above the

probe assembly (Figure 3.80). Plastic probe clips slide on with ﬁrm pressure.

Figure 3.80. Attaching a probe clip.

2. Install the pre-sterilized sensor and probe kit using aseptic

connection methods, as described in section 3.5.2. The aseptic connection is completed prior to the bellows being collapsed (Figure 3.81).

Figure 3.81. Installing the pre-sterilized sensor.

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Chapter 3 | Operating information

3. Insert the probe by collapsing the bellows (Figure 3.82).

Note: If the BPC is already ﬁlled with liquid, the best practice is to squeeze the bellows to expel air prior to collapsing it. Then insert the probe fully, as described.

Figure 3.82. Collapsed bellows.

4. Position the probe clip in the desired horizontal location. Lift the

probe and set it into the probe clip (Figure 3.83).

Figure 3.83. Lifting the probe upward into the spring.

5. Verify that the probe remains at the proper insertion depth and

angle as the bellows expand to rest freely in the probe clip.

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Chapter 3 | Operating information

3.5.4. Probe calibration

Probe calibration is controller-speciﬁc. However, the following general rules apply:

• pH probes must be calibrated prior to steam sterilization. The

calibration of the probe can be standardized by comparison of an off-line sample after the pH probe has been connected to the S.U.F.

• Dissolved oxygen probes are generally calibrated after steam

sterilization. After the probe is connected to the S.U.F. and is allowed sufﬁcient time to polarize (six to eight hours of continuous connection to the power supply provided by a controller or polarization module), it can be calibrated.

3.5.5. Kleenpak connection instructions

Use the following instructions to make Pall Kleenpak connections. This product is not sold sterile. For use in making sterile connections, each connector must be ﬁtted in a closed, single-use assembly that has been subjected to a validated sterilized process. It is very important that all instructions are carefully followed and where appropriate should be incorporated into the user’s standard operating procedures. If some of the procedures do not suit your needs, please consult Pall or your local distributor before ﬁnalizing your system.

Speciﬁcations

The Kleenpak connector has a maximum working pressure of 3 bar (43.5 psi) at 40°C in compatible ﬂuids.

WARNING: Operation outside the above speciﬁcations and/or with

ﬂuids incompatible with construction materials may cause personal injury and result in damage to the device.

Receipt of equipment

The male and female Kleenpak connectors are supplied in separate packages. There are several types of end ﬁttings in order to match different tubing size requirements and to allow for different attachment possibilities to ﬂexible tubing. See the Pall website for more information. Ensure the following upon receipt of equipment.

• Male and female connectors are supplied protected by an inner and

outer bag. Ensure that the packaging is undamaged.

• Store the male and female Kleenpak connectors in clean, dry

conditions and wherever practical in the external packaging as delivered.

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Chapter 3 | Operating information

• DO NOT remove from the inner device bag packaging until just

before installation.

• The assembly aid is provided non sterile and can be reused

multiple times. It needs to be stored in clean and dry conditions between each use. The assembly aid is supplied separately and is available for purchase from your local Pall representative.

Installation

Before installation, it is essential to verify that the connector is suitable for the liquid that it will be in contact with for the application and to follow the appropriate instructions listed below.

• Install the male and female connectors using compatible

connections. Ensure that the tubing is attached ﬁrmly to the hose barb to prevent leakage during operation using cable ties or other methods. During tubing assembly, premature actuation of the male plunger is prevented by the anti-actuation ring. The anti-actuation ring needs to remain in place until actual connection takes place. The presence of valves on the tubing before the connector is recommended to prevent liquid contact with the connectors prior to use.

• If the connectors are to be autoclaved, orient them with peel strips

facing upwards to prevent peel strip blockage by condensate.

WARNING: The device must remain dry prior to connection of

the male and female connectors. If there is ﬂuid present in the line or around the devices, do not use. If the protective cap has been removed, do not use.

Sterilization

WARNING: These disposable connectors must not be in-line steam

sterilized. Material design limitations will be exceeded when these devices are exposed to pressurized steam and they will be ruptured.

• Connect the male or female connector to the single use system to

be sterilized. A valve or clamp needs to be installed close to the connector to prevent accidental wetting after the system is ﬁlled with liquid.

• Ensure that the protective cap is ﬁrmly in place. Autoclave paper or

other radiation resistant material can be used to ensure that the cap does not become dislodged during handling.

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Chapter 3 | Operating information

Autoclave sterilization

• Install the male or female connector to the equipment to be

sterilized. If the connector is attached to a tank, the tank should be vented appropriately with a vent ﬁlter.

• Ensure that the protective cap of the connector is ﬁrmly in place.

Autoclave paper or other autoclavable and air/steam permeable material can be used to cover the cap loosely to ensure that the cap does not become dislodged during handling.

• The connectors should be allowed to vent during autoclaving. The

venting strip should be orientated upwards to prevent blockage by condensates.

CAUTION: To avoid collection of condensate within the

connectors, do not place the venting strip downwards during autoclaving.

• The connectors should not be covered with heavy objects during

the autoclave cycle.

Note: The maximum temperature is 121°C for ACD part numbers and 130°C for KPCHT part numbers and the maximum exposure time is 75 minutes. Do not autoclave at a higher temperature or for a longer time. A slow exhaust cycle is recommended.

WARNING: Do not autoclave the male and female connectors in the

bags that they are shipped in.

Important: Pall recommends that the efﬁciency of the sterilization cycle is validated using an appropriate method.

Making the connection

WARNING: Do not use if ﬂuid is in contact with the connector. Do not

use if protective caps are loose or displaced.

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Thumb rest

Anti-actuation ring

O-ring

Plunger

Barrel

Protective caps

Locking clips (on both sides)

FEMALE PART*

ACTUATED KLEENPAK STERILE CONNECTOR

MALE PART*

Base

Protective caps

Chapter 3 | Operating information

Making the connection using the connector assembly aid

See the next section of this guide for instruction about making the connection without using the connector assembly aid.

Figure 3.84. Kleenpak connector schematic.

1. Lift and pull tab off protective caps to remove caps from the

connectors (Figure 3.85).

Figure 3.85. Removing caps from connectors.

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Rounded side

Anti-actuation ring

Barrel

Female Connector

Flat side where both folded over peel away strips exit

Male Connector

Locking clips

(not visible)

Locking clips

Chapter 3 | Operating information

2. Hold the barrel of the larger (male) connector above the base.

• Align the smaller (female) connector with the larger (male) connector.

• Flat sides should be aligned.

• Both peel away strips need to remain folded (Figure 3.86).

Figure 3.86. Aligning male and female connectors.

Note: If the connectors are not aligned properly, the connection cannot be made.

3. Once aligned correctly, press the two connectors together ﬁrmly until both locking clips snap together tightly (Figure 3.87).

Figure 3.87. Pressing connectors together.

4. Support both the male and female connectors, and remove the anti-actuation ring from the male connector by pulling the tab towards the barbed end of the male connector (Figure 3.88).

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Remove the anti-actuation ring from the male connector

Assembly aid (cross section view)

Peel away strips

Chapter 3 | Operating information

Figure 3.88. Removing anti-actuation ring.

Note: Connector should stay securely in the assembly aid when properly installed.

Figure 3.89. Cross-section view of assembly aid.

5. Hold assembly aid in the palm of the hand with the connector facing outwards, and thumb supporting the connector in the assembly aid. Using the other hand, ﬁrmly grasp both peel away strips as close as possible to the body of the assembly aid to ensure a secure grip and pull both peel away strips simultaneously in a smooth continuous motion. Ensure that the connector is perpendicular to the peel away strips (Figure 3.90 and Figure 3.91).

Figure 3.90. Holding assembly aid with thumb supporting connector.

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Figure 3.91. Pulling peel away strips.

Chapter 3 | Operating information

WARNING: Do not use if only one peel away strip is removed

accidentally instead of two, this will affect the sterility of the pathway.

6. With the connector still secured in the assembly aid, push the thumb rest of the male connector down towards the base of the barrel (Figures 3.92 and 3.93).

Figure 3.92. Pushing down thumb rest of male connector.

Figure 3.93. Male connector pushed down.

Note that in order to establish a proper connection, the plunger inside the male connector must be fully inserted into the female connector. As a veriﬁcation, repeat actuation until a hard stop is reached.

If necessary, the connector may be removed from the assembly aid to complete the plunger movement.

7. Once the connector assembly is complete, the assembly aid may be removed. When assembly aid is removed, verify actuation until a hard stop is reached.

8. Start the ﬂuid transfer (Figure 3.94).

Figure 3.94. Starting ﬂuid transfer.

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Protective caps

Rounded side

Anti-actuation ring

Barrel

Female Connector

Flat side where both foldedover peel awaystrips exit

Male Connector

Chapter 3 | Operating information

Making the connection without using the connector assembly aid

1. Lift and pull tab off protective caps to remove caps from connectors (Figure 3.95).

Figure 3.95. Removing caps from connectors.

2. Hold the barrel of the larger (male) connector above the base.

• Align the smaller (female) connector with the larger (male) connector.

• Flat sides should be aligned.

• Both peel away strips need to remain folded (Figure 3.96).

Figure 3.96. Aligning male and female connectors.

Note: If the connectors are not aligned properly, the connection cannot be made.

3. Once aligned correctly, press the two connectors together ﬁrmly until both locking clips snap together tightly (Figure 3.97).

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Locking clips

(not visible)

Locking clips

Remove the anti-actuation ring from the male connector

Chapter 3 | Operating information

Figure 3.97. Pressing connectors together.

4. Support both the male and female connectors, and remove the anti-actuation ring from the male connector by pulling the tab towards the barbed end of the male connector (Figure 3.98).

5. With one hand, support the male and female sides of the Kleenpak

Note: Do not impart perpendicular forces on the connector, as it can cause the connector to break. If a perpendicular force is present due to items attached to the connector, then the connector must be properly supported.

Figure 3.98. Removing anti-actuation ring.

connector by wrapping ﬁngers around both sides of the connector, next to the ﬂange. Using the other hand, grasp both white peel away strips as close as possible to the ﬂat side of connector to ensure a good grip and pull them out simultaneously in a smooth continuous motion. Ensure that the connector is perpendicular to the peel away strips shown in shown in Figures 3.99 and 3.100. The perpendicular orientation must be maintained while the two strips are pulled simultaneously.

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Chapter 3 | Operating information

Figure 3.99. Holding assembly aid with thumb supporting connector.

Figure 3.100. Pulling peel away strips.

WARNING: Do not use if only one peel away strip is removed

accidentally instead of two, this will affect the sterility of the pathway.

6. Push the thumb rest of the male Kleenpak connector down towards the base of the barrel until they meet (Figures 3.101 and 3.102).

Figure 3.101. Pushing down thumb rest of male connector.

Figure 3.102. Male connector pushed down.

Note: In order to establish a proper connection, the plunger inside the male connector must be fully inserted into the female connector. As a veriﬁcation, repeat actuation until a hard stop is reached.

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Chapter 3 | Operating information

7. Start the ﬂuid transfer (Figure 3.103).

Figure 3.103. Starting ﬂuid transfer.

3.6. Microbial cell culture operating instructions

Before beginning your fermentation run, please refer to the checklist in Appendix D to ensure that all necessary steps have been completed.

3.6 .1. Operating conditions for microbial cell culture

applications

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

Table 3.2 lists values for both sizes of S.U.F.s. This data may be used in specifying maximum gas ﬂow rates for mass ﬂow controllers or rotameters. In optimal conditions (no condensation or fouling) the exhaust ﬁlters have a ﬂow capacity of at least 60 and 600 standard liters per minute (slpm) at 0–0.5 psig for the small and large standard equipped ﬁlter types, respectively. The total ﬂow rate of gas into system must be less than the sum ﬂow rate capacity of active exhaust ﬁlters. The values listed take into account the number and type of exhaust ﬁlters that are standard on each size of S.U.F. (one small ﬁlter installed in the 30 L, one large ﬁlter installed in the 300 L). These values are not absolute requirements. They are also not intended to be process gas ﬂow settings. The process gas ﬂow settings should be adjusted as discussed below with starting conditions not exceeding 25% of the listed maximum values to prevent unnecessary reduction of exhaust ﬁlter life span and foam generation.

If sparging seems to lose efﬁciency disproportionate to the cell culture density increase, more ﬁlters may be required or the oxygen to air ratio of the gas ﬂow may need to be increased.

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