GE WAVE Bioreactor 2 10 20 50 Systems Operating Manual PDF

GE Healthcare
WAVE Bioreactor™ 2/10

and 20/50 systems
Operating Instructions
Original instructions
Content
1 Introduction .............................................................................. 5
1.1 Important user information .................................................................................6
1.2 Regulatory information .........................................................................................7
1.3 WAVE system configurations .............................................................................9
1.4 WAVE Bioreactor 2/10 system ........................................................................ 11
1.5 WAVE Bioreactor 20/50 system ..................................................................... 12
1.6 Cellbag ........................................................................................................................ 13
1.7 WAVEPOD .................................................................................................................. 14
1.8 External instrument modules .......................................................................... 16
1.9 Control software .................................................................................................... 23
2 Safety instructions ................................................................ 25

2.1 Safety precautions ............................................................................................... 25
2.2 Labels .......................................................................................................................... 29
2.3 Emergency procedures ...................................................................................... 30
2.4 Recycling procedures .......................................................................................... 31
3 Installation.............................................................................. 33
3.1 Site requirements .................................................................................................. 33
3.2 Unpacking ................................................................................................................. 33
3.3 Installation of WAVE Bioreactor 2/10 .......................................................... 34
3.4 Installation of WAVE Bioreactor 20/50 ........................................................ 37
3.5 Installation of WAVEPOD .................................................................................... 41
3.6 Setting up external instrument modules .................................................... 45
3.7 Transport ................................................................................................................... 47
3.8 Spare parts and accessories ............................................................................ 47
4 Control systems..................................................................... 49
4.1 WAVE Bioreactor 2/10 control system ........................................................ 49
4.2 WAVE Bioreactor 20/50 control system ..................................................... 56
4.3 WAVEPOD control system ................................................................................. 66
4.4 External instrument module control systems ......................................... 83
5 Operation ................................................................................ 95
5.1 Operation overview .............................................................................................. 95
5.2 Preparations before starting a culture ........................................................ 96
5.3 Performing a run ..................................................................................................102
5.4 Procedures after a run ......................................................................................109
6 Maintenance......................................................................... 111
6.1 General .....................................................................................................................111
6.2 Safety switch inspection procedure ...........................................................112
6.3 Replacement of fuses ........................................................................................113
6.4 Calibration ..............................................................................................................114
WAVE Bioreactor 2/10 and 20/50 systems Operating Instructions 28-9641-23 AA 3

7 Troubleshooting .................................................................. 117
7.1 WAVE Bioreactor 2/10 ...................................................................................... 117
7.2 WAVE Bioreactor 20/50 ................................................................................... 121
7.3 WAVEPOD ............................................................................................................... 128
7.4 External instrument modules ........................................................................ 133
8 Reference information ....................................................... 139

8.1 Specifications ....................................................................................................... 139
8.2 Literature ................................................................................................................ 142
8.3 Ordering information ........................................................................................ 143
4 WAVE Bioreactor 2/10 and 20/50 systems Operating Instructions 28-9641-23 AA

Introduction 1
1 Introduction
Purpose of the Operating
Instructions
The Operating Instructions provide you with the instructions needed to handle WAVE
Bioreactor 2/10 and 20/50 systems in a safe way.
Prerequisites
In order to operate WAVE Bioreactor 2/10 and 20/50 systems safely and according to
the intended purpose the following prerequisites must be met:
• You should be acquainted with the use of general laboratory equipment and with
handling of biological materials.
• You must read the Safety Instructions in Chapter 2 of these Operating Instructions.
• The system should be installed according to the instructions in Chapter 3 of these
Operating Instructions.
In this chapter
This chapter contains important user information and a general description of
WAVE Bioreactor 2/10, WAVE Bioreactor 20/50, WAVEPOD™, and external instrument
modules, and their intended use.

1 Introduction
1.1 Important user information
1.1 Important user information

Read this before using WAVE
All users must read the Safety Instructions in Chapter 2 of these Operating Instructions
before installing, using or maintaining the system.
Do not operate WAVE Bioreactor 2/10 and 20/50 systems in any other way than
described in the user documentation. If you do, you may be exposed to hazards that
can lead to personal injury and you may cause damage to the equipment.
Intended use
The WAVE Bioreactor is a cell culture device. Culture medium and cells are loaded in to
a single use, presterilized bag known as a Cellbag™. The Cellbag is then placed on an
electric rocking base, the rocking unit.
WAVE Bioreactor 2/10 and 20/50 systems shall not be used in any clinical procedures,
or for diagnostic purposes.
Safety notices
These Operating Instructions contain WARNINGS, CAUTIONS and NOTICES concerning
the use of the product, with meanings as defined below.
WARNING
WARNING indicates a hazardous situation which, if not avoided, could
result in death or serious injury. It is important not to proceed until all
stated conditions are met and clearly understood.
CAUTION
CAUTION indicates a hazardous situation which, if not avoided, could
result in minor or moderate injury. It is important not to proceed until all
stated conditions are met and clearly understood.
NOTICE
NOTICE indicates instructions that must be followed to avoid damage to
the product or other equipment.

Introduction 1
Regulatory information 1.2
Notes and tips

Note: A Note is used to indicate information that is important for trouble-free and
optimal use of the product.
Tip: A tip contains useful information that can improve or optimize your procedures.
Typographical conventions
Software texts and commands are identified by bold italic text. A colon is used to
separate menu levels (e.g., File:Open refers to the Open option in the File menu).
1.2 Regulatory information

This section lists the directives and standards that are fulfilled by WAVE Bioreactor 2/10
and 20/50 systems.
Manufacturing information
Requirement Content
Name and address of manufacturer GE Healthcare Bio-Sciences AB,

Björkgatan 30, SE 751 84 Uppsala
Sweden
Name and ID of notified body INTERTEK SEMKO AB, NB 0413
Place and date of declaration Uppsala, Sweden, Nov 2009
Identity of person authorized to sign DoC See EC Declaration of Conformity.
CE Conformity
Directive Title
2006/42/EC Machinery Directive (MD)
2006/95/EC Low Voltage Directive (LVD)
2004/108/EC ElectroMagnetic Compatibility (EMC) Directive

1 Introduction
1.2 Regulatory information
International standards
Standard Description Notes
EN 61010-1, Safety requirements for electrical

IEC 61010-1, equipment for measurement, control
CAN/CSA-C22.2 no. and laboratory use
61010-1
EN 61326-1 EMC emissions and immunity Harmonized with

requirements for measurement, 2004/108/EC
control and laboratory use
EN-ISO Safety of machinery – Basic concepts, Harmonized with

12100-1, 12100-2 general principles and design 2006/42/EC
EN-ISO Safety of machinery – Principles of Harmonized with

14121-1, 14121-2 risk assessment 2006/42/EC
CE marking
The CE marking and the corresponding Declaration of Conformity is valid for the
instrument when it is:
• used as a stand-alone unit, or
• connected to other CE-marked instruments, or

• connected to other products recommended or described in the user
documentation, and
• used in the same state as it was delivered from GE Healthcare, except for
alterations described in the user documentation or explicitly authorized by
GE Healthcare.
Regulatory compliance of
connected equipment
Any equipment connected to WAVE Bioreactor 2/10 system and WAVE Bioreactor 20/
50 system should meet the safety requirements of EN 61010-1/IEC61010-1 or relevant
harmonized standards. Within the European Union, connected equipment must be
CE-marked.

Introduction 1
WAVE system configurations 1.3
1.3 WAVE system configurations

The WAVE Bioreactor systems provide scalable cell culturing possibilities with the
control of key parameters. The WAVE Bioreactor systems are available in several
different configurations allowing control of some or all of these key parameters.
The parameters can be controlled by instrument modules integrated into the WAVE
Bioreactor and/or by external instrument modules, that is, WAVEPOD (available with
different functionalities) and standalone modules.
For examples of how control of different parameters can be achieved, see Table 1-1.
Note that more than one external instrument module can be used with a WAVE
Bioreactor base unit to get access to more control functions.
For some parameters, several control options are available. For example, pH in the
culture medium can be controlled by:
• On-line control of pH by pH measurement and pH adjustment by acid and base
additions. This requires:
• WAVEPOD with pH control module and PUMP20, or
• the external instrument modules pH20 and PUMP20.
• Indirect control of pH by control of the CO2 concentration. This requires:
• Integrated CO2 control module (as in BASE20/50EHT-CO2), or
• WAVEPOD with CO2 control module, or
• the external instrument modules CO2MIX20 or CO2MIX20-R.

1 Introduction
1.3 WAVE system configurations
Table 1-1. Examples of WAVE Bioreactor system configurations. Note that

WAVE Bioreactor 20/50 system is available also as Dual system configuration (WAVE
Bioreactor 20/50-D) which enables operation of two Cellbags in parallel.
System configuration Control parameters
CO2 concentration
Perfusion culture
O2 concentration
(measurement)
Temperature
Dissolved O2
pH control
External
Rocking
Bioreactor instrument
base unit module
WAVE Bioreactor 2/10 system
BASE2/10EH + +
BASE2/10EH DOOPT20 + + +
BASE2/10EH pH20 and PUMP20 + + +
BASE2/10EH CO2MIX20 + + +
BASE2/10EH O2MIX20 + + +
BASE2/10EH PERFCONT2E + + +
WAVE Bioreactor 20/50 system
BASE20/50EHT + +
BASE20/50EHT-CO2 + + +
BASE20/50EHT-O2 + + +
BASE20/50EHT-L + + +
BASE20/50EHT POD-R/L PH CO2 + + + +
BASE20/50EHT POD-R/L DOOPT O2 + + + +
BASE20/50EHT POD-R/L PH DOOPT CO2 O2 + + + + + +
BASE20/50EHT-L POD-R/L PH DOOPT CO2 O2 + + + + + + +
BASE20/50EHT DOOPT20 + + +
BASE20/50EHT pH20 and PUMP20 + + +
BASE20/50EHT CO2MIX20 + + +
BASE20/50EHT O2MIX20 + + +

Introduction 1
WAVE Bioreactor 2/10 system 1.4
1.4 WAVE Bioreactor 2/10 system

Introduction
A complete WAVE Bioreactor 2/10 system consists of:
• WAVE Bioreactor 2/10
• Cellbag holder
• Pre-sterilized disposable Cellbag (ordered separately)
• Perfusion controller (optional)
• External instrument modules (optional)
Illustration of
WAVE Bioreactor 2/10
The illustration below shows the main parts of the WAVE Bioreactor 2/10.
4
3
2
6
1
Part Description
1 Bioreactor base unit
2 Front panel with controls
3 LCD display
4 Rocking unit
5 Cellbag holder
6 Perfusion controller (optional)

1 Introduction

Introduction
A complete WAVE Bioreactor 20/50 system consists of:
• WAVE Bioreactor 20/50
• Cellbag holder
• Pre-sterilized disposable Cellbag (ordered separately)
• WAVEPOD (optional)
• External instrument modules (optional)
Illustration of
The illustration below shows the main parts of the WAVE Bioreactor 20/50.
Part Description
1 Bioreactor base unit
2 Protective bellows, rocking mechanism
3 Rocking unit
4 Cellbag holder (one of several options shown)
5 Detachable Touch screen

Introduction 1
Cellbag 1.6
1.6 Cellbag
Introduction
The Cellbag is a disposable bioreactor chamber. It is presterilized by gamma radiation
and is intended for single use. The Cellbag is partially filled with culture media,
inoculated, and placed on the bioreactor base unit.
Illustration
The illustration below shows the location of the main parts of the Cellbag.
" #
&
Part Description
1 Cellbag rod
2 Inoculation/harvest lines
3 Outlet air filter
4 Inlet air filter
5 Oxywell2™
6 Needleless sampling port
7 Spare Luer port/optional pH probe

1 Introduction
1.7 WAVEPOD
1.7 WAVEPOD
Introduction
The WAVEPOD instrument module integrates the instrumentation associated with
WAVE Bioreactor 20/50. This includes pH, dissolved oxygen, and CO2/O2 gas mixing
controls. Up to four of instrument modules can be installed into WAVEPOD at the same
time to provide desired functionality. The WAVE Bioreactor can be operated from the
WAVEPOD Touch screen.
The WAVEPOD can be placed on either side of a WAVE Bioreactor and is available in
both left and right facing versions.
Available WAVEPOD control

modules
The following modules can be integrated into WAVEPOD.
Module Description
pH controller Enables on-line control of pH in the Cellbag
DO (Dissolved Oxygen) Enables on-line control of dissolved oxygen in the

controller Cellbag
Airflow controller Controls the flow of air through the Cellbag. The Airflow
controller is required if the CO2 or O2 controllers are
used
CO2 controller Measures and controls the CO2 concentration in the

Cellbag
O2 controller Measures and controls the O2 concentration in the

Cellbag

Introduction 1
WAVEPOD 1.7
Illustration
The illustration below shows the location of the main parts of WAVEPOD.
3
5
2
Part Description
1 Bay 1 - pH module
2 Bay 2 - DO module
3 Bay 3 - airpump module
4 Bay 4 - CO2 and O2 modules
5 Touch screen

1 Introduction
1.8 External instrument modules

Overview
Several external instrument modules for the WAVE Bioreactor 2/10 and 20/50 systems
are available. The external instrument modules are used as standalone modules
connected to the bioreactor.
The available external instrument modules are listed in the table below.
Name Function
DOOPT20 Dissolved oxygen monitor
CO2MIX20 CO2/air controller
CO2MIX20-R CO2/air controller (rotameter model)
O2MIX20 O2/air controller
O2MIX20-R O2/air controller (rotameter model)
PUMP20 Peristaltic feed/harvest or acid/base pump
pH20 Acid/base controller

Introduction 1
External instrument modules 1.8
DOOPT20 dissolved oxygen

monitor
DOOPT20 is used to monitor dissolved oxygen (DO) levels in the Cellbags. In the WAVE
Bioreactor application, the DOOPT probe is used in the Oxywell2 configuration.
Oxywell2 is a silicone rubber sleeve that is provided inside the Cellbag. The DOOPT
probe is inserted into the sleeve and oxygen is measured non-invasively as it diffuses
through the silicone rubber.
The illustration below shows the location of the main parts of DOOPT20.
3
4
5
Part Description Part Description
1 Process controller 4 TEMP COMP control
2 LCD display 5 PUSH FOR MENU button
3 Probe connector

1 Introduction
CO2MIX20 CO2/air controller

CO2MIX20 is used to inflate and ventilate the Cellbags with air containing a variable
carbon dioxide (CO2) concentration. CO2 from a pipeline or cylinder is connected to
CO2MIX20. Air is drawn from the AIR IN port and mixed with the CO2 (connected to CO2
IN). A CO2 sensor measures the CO2 concentration and controls it to any user-specified
value from 0% to 15%. The CO2 conditioned air is pumped from the MIX OUT port into
the headspace of the Cellbag to maintain bag pressure, and to provide oxygen and
carbon dioxide for ventilation and pH control.
The illustration below shows the location of the main parts of CO2MIX20.
1 2 3
4
8 5
7 6
1 Airflow controller 6 CO2 PRESSURE OK LED
2 HIGH PRESSURE LED 7 CO2 IN port
3 PUMP ON switch 8 AIR IN port
4 Process controller 9 MIX OUT port
5 CO2 ON switch

Introduction 1
CO2MIX20-R CO2/air controller

CO2MIX20-R (rotameter model) is used to inflate and ventilate the Cellbags with air
containing a variable carbon dioxide (CO2) concentration. CO2 from a pipeline or
cylinder is connected to the CO2MIX20-R, which has a precision regulator to adjust the
gas pressure. Air is drawn from the AIR IN and mixed with the CO2 (connected to
CO2 IN). A CO2 sensor measures the CO2 concentration and controls it to any user-
specified value from 0% to 15%. The CO2 conditioned air is pumped from the MIX OUT
port into the headspace of the Cellbag to maintain bag pressure, and to provide oxygen
and carbon dioxide for ventilation and pH control.
The illustration below shows the location of the main parts of CO2MIX20-R.
1 2
10 9 8 7 6 5
1 Process controller 6 Pressure regulator
2 CO2 gauge 7 AIR IN port
3 Airflow rotameter 8 PUMP ON switch
4 Flow rate adjust 9 CO2 IN port
5 MIX OUT port 10 CO2 ON switch

1 Introduction
O2MIX20 O2/air controller

O2MIX20 is used to inflate and ventilate the Cellbags with air containing a variable O2
concentration. O2 from a pipeline or cylinder is connected to O2MIX20. Air is drawn from
the AIR IN port and mixed with the O2 (connected to O2 IN). An O2 sensor measures the
O2 concentration and controls it to any user-specified value from 21% to 50%. The O2
conditioned air is pumped from the MIX OUT port into the headspace of the Cellbag to
maintain bag pressure, and to provide oxygen.
The illustration below shows the location of the main parts of O2MIX20.
1 2
3
4
5
6
9 8
1 Process controller 6 AIR IN port
2 Airflow controller 7 O2 PRESSURE OK LED
3 PUMP ON switch 8 O2 IN port
4 HIGH PRESSURE LED 9 O2 ON switch
5 MIX OUT port

Introduction 1
O2MIX20-R O2/air controller

O2MIX20-R (rotameter model) is used to inflate and ventilate Cellbags with air
containing a variable O2 concentration. O2 from a pipeline or cylinder is connected to
O2MIX20-R which has a precision regulator to adjust the gas pressure. Air is drawn from
the AIR IN port and mixed with the O2 (connected to O2 IN). An O2 sensor measures the
O2 concentration and controls it to any user-specified value from 21% to 50%. The O2
conditioned air is pumped from the MIX OUT port into the headspace of the Cellbag to
maintain bag pressure, and to provide oxygen.
The illustration below shows the location of the main parts of O2MIX20-R.
1 2
9 8 7 6 5
1 Process controller 6 Pressure regulator
2 O2 gauge 7 AIR IN port
3 Airflow rotameter 8 PUMP ON switch
4 Flow rate adjust 9 O2 IN port
5 MIX OUT port 10 O2 ON switch

1 Introduction
PUMP20 peristaltic feed/

harvest pump
PUMP20 is a peristaltic pump. It is mainly intended for use as a feed/harvest pump for
use with the WAVE Bioreactors. It can also be used for acid/base additions. PUMP20 can
be placed on either side of a WAVE Bioreactor and is available in two versions,
PUMP20-L facing left and PUMP20-R facing right.
The illustration below shows the location of the main parts of PUMP20.
1
3
1 PUMP ON switch 3 Digital speed control buttons
2 RUN switch

Introduction 1
Control software 1.9
pH20 acid/base controller

pH20 acid/base controller enables on-line control of pH in a Cellbag. A setpoint
controller is provided to power base and acid pumps on and off to maintain pH. pH20
has DB9 connectors to remotely switch PUMP20 units or user-supplied pumps.
The pH20 requires special pH probes made by GE Healthcare that can be inserted into
a luer fitting provided on the Cellbag. These pH probes are provided sterile and must be
inserted in an aseptic manner.
The illustration below shows the location of the main parts of pH20 acid/base controller.
4
3
5
2
7 6
1 pH sensor connector 5 Up/down arrow buttons
2 SLOPE knob 6 BASE ENBL switch
3 SPAN knob 7 ACID ENBL switch
4 Process controller
1.9 Control software

The WAVE instruments are equipped with embedded software for control and
supervision. For information on how to connect WAVE instruments to external software,
refer to WAVE Bioreactor System BASE 2/10 EH Operator Manual or WAVE Bioreactor
System 20/50EHT Operator Manual.

1 Introduction
1.9 Control software

Safety instructions 2
Safety precautions 2.1
2 Safety instructions
This chapter describes safety compliance, safety labels, general safety precautions,
emergency procedures, power failure and recycling of WAVE instruments.
2.1 Safety precautions

Introduction
Before installing, operating of maintaining the system, you must be aware of the
hazards described in the user documentation. Follow the instructions provided to avoid
personal injury or damage to the equipment.
The safety precautions in the section are grouped into the following categories:
• General precautions
• Personal protections
• Installing and moving the instrument
• System operation
• Maintenance
General precautions
WARNING
Do not operate the WAVE systems in any other way than described in
the WAVE user documentation.
WARNING
Operation and user maintenance of the WAVE system should be
performed by properly trained personnel only.
WARNING
Do not use any accessories not supplied or recommended by GE
Healthcare.

WARNING
Only WAVE approved Cellbags shall be used on the equipment.
Using flammable liquids
WARNING
The WAVE Bioreactors are not designed to handle flammable fluids. The
WAVE Bioreactor are not approved for work in a potentially explosive
atmosphere.
Personal protection
WARNING
To avoid hazardous situations when working with the WAVE systems,
take the following measures for personal protection.
WARNING
Always use appropriate personal protective equipment during
operation and maintenance of WAVE systems.
WARNING
Hazardous substances. When using hazardous chemical and
biological agents, take all suitable protective measures, such as
wearing protective glasses and gloves resistant to the substances used.
Follow local and/or national regulations for safe operation and
maintenance of the system.
WARNING
Spread of biological agents. The operator has to take all necessary
actions to avoid spreading hazardous biological agents in the vicinity of
the instrument. The facility should comply with the national code of
practice for biosafety.

Safety precautions 2.1
Installing and moving the

instrument
WARNING
Emergency stop. Position the WAVE system so that the power switch is
easily accessible for power shut off. The power switch is located at the
rear side of the instrument.
WARNING
Protective ground. The WAVE instruments must always be connected
to a grounded power outlet.
WARNING
Heavy object. Filled Cellbags have a considerable weight and heavy
lifts must be done with care. Use 1 person per multiple of 15 kg weight,
for example 3 persons for 30 to 45 kg. All lifting and moving must be
performed in accordance with local regulations.
CAUTION
Ensure that all tubing, hoses and cables are placed so that the risk or
tripping accidents is minimized.
CAUTION
Make sure that there is enough free space around the instrument for
the rocking motion.
CAUTION
The safety switches of the base unit must be functionally tested after
the installation of the equipment or after the instrument has been
transported and every 6 months from then on. See Chapter 6 for proper
method of testing.
System operation
WARNING
Biohazard. Make sure that the Cellbag is sealed before and during the
fermenting process.

WARNING
Electrical shock hazard after spillage. If there is a risk that large
volumes of spilled liquid may penetrate the casing of the WAVE
instrument, immediately switch off the instrument, disconnect the
power cord, and contact an authorized service engineer.
CAUTION
Pinch hazard. The rocking unit is a safety hazard. Remain clear of all
moving parts during operation. Do not work on the bag or rocking unit
unless the rocking motion is stopped.
CAUTION
Overheating is possible if the unit is operated without a liquid-
containing bag. The Cellbag holder temperature may exceed 60°C. Do
not touch.
CAUTION
Leakage risk of biological substances. Before every use, check all
hoses for signs of cracking or tears. None of the air hoses should
contain liquids of any kind.
CAUTION
Remove any spillage on the floor immediately to minimize the risk for
slipping accidents.
CAUTION
Pinch hazard when using PERFCONT. Do not run the feed and harvest
pump with the door open
Maintenance
WARNING
Electrical shock hazard. All repairs should be done by service
personnel authorized by GE Healthcare. Do not open any covers or
replace parts unless specifically stated in the user documentation.
WARNING
Disconnect power. Always disconnect power from the instrument
before performing any maintenance task.

Labels 2.2
WARNING
Always clean the equipment a well ventilated area. Never douse or
immerse any part of the unit with any liquid. When cleaning is required,
use only water and alcohol.
WARNING
Only spare parts that are approved or supplied by GE Healthcare may
be used for maintaining or servicing the system.
2.2 Labels
This section describes safety labels and labels concerning hazardous substances that
are attached to the WAVE instruments.
Labels on the instrument

The illustration below shows an example of the identification label that is attached to
the WAVE instruments.
28941342 BASE20/50EHT 220-240V

Code no: 28942137 Voltage:220-240
Serial no: 1234567 Frequency: 50/60 Hz
Mfg Year: 2009 Max Power: 630 VA
Fuse: 2x T 6.3AL 250 V
Made in Sweden GE Healthcare Bio-Sciences AB

751 84 Uppsala Sweden
Symbols used in safety labels
Label Description
Warning! Read the user documentation before using the system. Do

not open any covers or replace parts unless specifically stated in the
user documentation.
The system complies with the requirements for electromagnetic

compliance (EMC) in Australia and New Zealand.
The system complies with applicable European directives.

2.3 Emergency procedures
Labels concerning hazardous

substances
Label Description
This symbol indicates that the waste of electrical and electronic

equipment must not be disposed as unsorted municipal waste and
must be collected separately. Please contact an authorized
representative of the manufacturer for information concerning the
decommissioning of equipment.
This symbol indicates that the product contains hazardous materials

in excess of the limits established by the Chinese standard SJ/
T11363-2006 Requirements for Concentration Limits for Certain
Hazardous Substances in Electronics.
2.3 Emergency procedures

This section describes how to do an emergency shutdown of a WAVE instrument. The
section also describes the result in the event of power failure
Emergency procedures
In an emergency situation, do as follows to stop the run:
Step Action
1 Switch off the power to the instrument by pressing the power switch to
the O position.
2 If required, disconnect the power cord from the power outlet.
Power failure
In the event of power failure, the run is immediately interrupted. If the AUTOSTART
option is set to ON, the operation is automatically resumed on power up. For further
information, please refer to the user documentation.
Safety switches
Safety switches are located on top of the bioreactor base units. If any of the safety
switches is hit, the rocking motion stops and rocking unit moves to a level position. If a
safety switch has been hit, power must be switched off and then on again to reset the
safety switch.

Recycling procedures 2.4
2.4 Recycling procedures

The equipment shall be decontaminated before decommissioning and all local
regulations shall be followed with regard to scrapping of the equipment.
Disposal, general instructions

When taking WAVE systems out of service, the different materials must be separated
and recycled according to national and local environmental regulations.
Recycling of hazardous
substances
WAVE systems contain hazardous substances. Detailed information is available from
your local GE Healthcare representative.
Disposal of electrical
components
Waste of electrical and electronic equipment must not be disposed as unsorted
municipal waste and must be collected separately. Please contact an authorized
representative of the manufacturer for information concerning the decommissioning of
your equipment.

2.4 Recycling procedures

Installation 3
Site requirements 3.1
3 Installation
This chapter provides information regarding installation of WAVE Bioreactor 2/10,
WAVE Bioreactor 20/50, WAVEPOD, and external instrument modules.
For further information on installation, refer to the user documentation listed in
Section 8.2.
3.1 Site requirements

Parameter Requirement
Electrical power 110 to 120 V~/ 220 to 240 V~,

50/60 Hz
Placement Stable laboratory bench
Operating temperature, 0ªC to 50ªC

WAVE Bioreactor 2/10 and WAVE Bioreactor 20/50
Operating temperature, WAVEPOD 5ªC to 40°C
Humidity, <95%, non-condensing

WAVE Bioreactor 2/10 and WAVE Bioreactor 20/50
Humidity, WAVEPOD 10% to 90%, non-condensing
3.2 Unpacking
Unpack the equipment and place it on a stable surface. If applicable, adjust the leveling
feet.
Check the equipment for any apparent damage before starting installation. Document
any damage carefully and contact your GE Healthcare representative.

3 Installation
3.3 Installation of WAVE Bioreactor 2/10

WAVE Bioreactor 2/10 rear
panel
1 2 3 4 5 6 7
9 8
1 Power connector 6 TEMP connector
2 Voltage selector 7 MODBUS connectors
3 Power switch 8 SCALE connector
4 HEATER connector 9 FILTER HEATER connector
5 ALARM connector

Installation 3
Installation of WAVE Bioreactor 2/10 3.3
Installing PERFCONT2E
perfusion module (optional)
An optional perfusion module is available for the WAVE Bioreactor 2/10. This modules
provides weight-based control of feed and harvest.
Follow the instruction below to install the perfusion module.
Step Action
1 Place the perfusion module on a stable surface. Adjust the levelling feet.
2 Place the WAVE Bioreactor 2/10 on top of the perfusion module. Make
sure all four feet are properly placed on the stainless-steel platform.
3 Connect the supplied PERFcable (DB9-F) to the WAVE Bioreactor 2/10 rear
panel port marked SCALE and to the rear of the perfusion module. Secure
the cable at both ends using the thumbscrews.
4 Connect tubing from the feed and harvest containers to the Cellbag.
Make sure the flow direction corresponds to the arrows marked on the
peristaltic pumps and that the tubing is properly pinched in the pump
rollers.
5 Open any clamps on the feed and harvest lines. Swing the clear covers to
the right on the pump override buttons, and press the buttons to test and
prime the pumps.
Setting up the bioreactor

Follow the instruction below to set up the bioreactor.
Step Action
1 Place the instrument on a stable surface.
2 Place the Cellbag holder on top of the stainless-steel rocker with the
heater cable towards the rear of the rocker.
3 Plug the blue heater cable into the rear panel connector labeled HEATER.
Twist the lock nut to secure.
4 Plug the metal end of the yellow temperature sensor cable into the
connector on the rear panel labeled TEMP. Twist the metal collar to lock
the plug. Plug the other end of the cable to the temperature sensor
attached to the Cellbag holder.
5 Check that the voltage selector is set to correct voltage. If not, pry out the
jumper using a screwdriver, adjust the arrow so that it is pointing to the
desired line voltage, and reinsert the jumper.

3 Installation
Step Action
6 Connect the supplied electrical cord to the power inlet and to a power
outlet.
7 Press the power switch on the rear panel to the I position to turn on the
instrument.
Result: The LCD screen lights up and the instrument starts. The instrument
tilts to its home position and when initialization is completed the main
screen is displayed.
Setting up the aeration system

of the bioreactor
The Cellbags require aeration to keep them inflated and to provide ventilation. The
ventilation air can be room air, incubator air, or a special gas mixture depending on the
cell line and buffer system in use.
Follow the instruction below to set up the aeration system of the bioreactor.
Step Action
1 Connect the supplied air tube between the AIR OUT port located on the
left side of the bioreactor and the inlet filter of the Cellbag.
2 If ambient air is to be used for aeration, leave the AIR IN port

unconnected. Otherwise, connect the desired CO2/air mixture to the AIR
IN port. CO2MIX20 controllers can be used to produce any required CO2
gas mixture.
NOTICE
Pressure at the AIR IN port must not exceed 1 psig. Exceeding this limit
may cause rupture of the Cellbag. Provide a suitable safety pressure
relief device.

Installation 3

WAVE Bioreactor 20/50 rear
panel
The illustration below shows the rear panel of WAVE Bioreactor 20/50.
10
1 2 3 4 5 6 7 8 9
1 DATAPORT 1 and 2 6 FILTER HEATER connector
2 FEED PUMP connector 7 HEATER connector
3 HARVEST PUMP connector 8 Power connector
4 ALARM ANALOG OUT 9 Power switch

connector
5 LOAD CELL connector 10 Fuse holder

3 Installation
WAVE Bioreactor 20/50 side

panel
The illustration below shows the side panel of WAVE Bioreactor 20/50.
1 Temp connector 3 Air in connector
2 Air out connector
Installation of KIT20EHT(D) or
KIT50EHT(D)
KIT20EHT(D) or KIT50EHT(D) are used with the WAVE Bioreactor 20/50 rocking unit. Each
kit includes the hardware required to properly mount a Cellbag holder and a Heater pad
to control the operating temperature.
Follow the instruction below to install KIT20EHT(D) or KIT50EHT(D).
Step Action
1 Place the Top plate onto the bioreactor base unit and align the four
mounting holes. Make sure that the opening for cables is on the left when
facing the front of the bioreactor.
2 Fasten the Top plate by screwing the four mounting screws through the
mounting holes using an Allen key.

Installation 3
Setting up the temperature

control system
Follow the instruction below to set up the temperature control system.
Step Action
1 Place the Heater pad on the Top plate. Make sure that the Heater pad is
oriented with the white side up and with the power connector fitting into
the slot on the left side of the Top plate.
2 Plug the blue heater cable into the rear panel connector labeled HEATER.
3 Place the stainless steel holder on top of the Heater pad and snap it in.
4 Place the SRTDX temperature sensor in the SRTDX retainer
5 Connect the yellow temperature sensor cable to the Temp connector on

the left side of the stainless steel holder. Connect the other end of the
cable into the temperature sensor connector on the left side panel. Screw
the locking nuts to firmly secure the plugs.
6 Check that the temperature display on the main screen reads the
ambient temperature.
7 For dual-sided system: Connect another yellow temperature sensor cable

to the temperature sensor connector on the left side of the Cellbag
holder. Connect the other end of the cable into the SRTDX temperature
sensor connector on the right side panel. Screw the locking nuts to firmly
secure the plugs.
Connecting power
Follow the instruction below to connect power to the bioreactor.
Step Action
1 Connect the supplied power cord to the power inlet and to a grounded
power outlet.
2 Press the power switch on the rear panel to the I position to turn on the
instrument.
Result: The Touch screen lights up and the instrument starts. The
instrument tilts to its home position and when initialization is completed
the main screen is displayed.

3 Installation
Setting up the aeration system

of the bioreactor
The Cellbags require aeration to keep them inflated and to provide ventilation. The
ventilation air can be room air, incubator air, or a special gas mixture depending on the
cell line and buffer system in use. The WAVE Bioreactor 20/50 has an internal air pump
(two in the dual version) that is used to aerate the Cellbags.
Follow the instruction below to set up the aeration system of the bioreactor.
Step Action
1 Connect the supplied air tube between the Air out port located on the
side panel of the bioreactor and the inlet filter of the Cellbag.
2 If ambient air is to be used for aeration, leave the Air in port

unconnected. Otherwise, connect the desired air mixture to the Air in
port. CO2MIX20 controllers can be used to produce any required CO2 gas
mixture.
Setting the real-time clock

All alarm data is labeled with the current date and time. It is therefore important to set
the clock to the local time.
Follow the instruction below to set the real time clock.
Step Action
1 On the main screen, press MENU.
2 On the MENU screen, press SETUP.
3 Enter the password “2050”.
4 On the SETUP screen, press SET CLK.
5 Change the date and time as needed. Press ↵ to accept each entry.
6 Press SET to accept the new date and time or BACK to abort.
Result: The main screen is displayed.
Setting up LCELL20/50EHT
optional weight sensor
The optional weight sensor is mounted under the Cellbag holder and provides the
weight of the Cellbag holder and the Cellbag.
To connect the LCELL20/50EHT to the WAVE Bioreactor 20/50, connect the yellow 4-pin
cable from the LCELL20/50EHT to the connector labeled LOAD CELL located on the rear
panel of the bioreactor.

Installation 3
Installation of WAVEPOD 3.5
Setting up the Cellbag bar code

detector
Each Cellbag holder is equipped with a bar code bag detector. The bar code bag
detector senses the presence of a bar code that is affixed to every Cellbag. The bar code
detector verifies that the Cellbag is a WAVE licensed product and reads the serial
number.
To set up the bar code detector, connect the yellow cable from the bar code detector
located under the Cellbag holder to the connector located on top of the WAVE
Bioreactor 20/50.
3.5 Installation of WAVEPOD

WAVEPOD should be physically installed next to the WAVE Bioreactor or as close as
possible.
Equipment required
The items listed below are recommended to facilitate the installation of WAVEPOD.
Some items are facility sources and other items can be purchased from GE Healthcare.
Facilities
• Oxygen (O2) supply source - pressure regulated to 0.7 to 1 bar (10 to 15 psig)
• Carbon dioxide (CO2) supply source - pressure regulated to 0.7 to 1 bar (10 to 15
psig)
Equipment and materials

• Acid pump - PUMP20 or customer supplied pump
• Base pump - PUMP20 or customer supplied pump
• pH probe
• DOOPT probe
• CIDEX disinfectant solution for pH probes

3 Installation
Connecting power
Follow the instruction below to connect power to WAVEPOD.
Step Action
1 Connect WAVEPOD to a grounded power outlet. The power connector is

located on the rear of the instrument.
2 Turn on the power to WAVEPOD by pressing the power switch located on

the rear panel to the I position.
Result: The Ethernet indicator lights up and a message is displayed
stating: BASE not found. This message is displayed until a WAVE
Bioreactor is connected to WAVEPOD. After a few seconds the overview
screen is displayed.
Connecting a bioreactor base

unit
Follow the instruction below to connect WAVEPOD to a WAVE bioreactor base unit.
Step Action
1 Connect the WAVEPOD-base unit interconnection cable to the port

labeled BASE20/50EHT located on the rear panel of WAVEPOD.
2 Connect the other end of the interconnection cable to the connector

labeled DATAPORT 1 located on the rear panel of the WAVE Bioreactor
base unit.

Installation 3
Installation of WAVEPOD 3.5
Step Action
3 On the WAVEPOD Touch screen, navigate to the SETUP screen. Starting

from the overview screen, press MENU to display the SYSTEM INFO
screen, and then press SETUP to display the SETUP
screen.
4 Enter the password (default password is “2050”).
5 On the SETUP screen of the WAVEPOD Touch screen, press OPTIONS.

Result: The OPTIONS screen is displayed.
6 On the OPTIONS screen, select the type of base unit.

3 Installation
Step Action
7 Switch off WAVEPOD and then switch it on again to initiate

communication with the connected bioreactor base unit.
Result: After 20 seconds the SYSTEM INFO screen is displayed. Then the
overview screen with the data of the connected base unit is displayed.
Connecting acid/base pumps

Two DB9 connectors for connection of PUMP20 acid and base pumps are located on the
rear panel of WAVEPOD. Connect the pumps as described in the table below. Use the
cables supplied with the pump modules.
Connect the... to the connector marked...
acid pump PUMP1
base pump PUMP2

Installation 3
Setting up external instrument modules 3.6
3.6 Setting up external instrument modules

For further information on how to set up the external instrument modules, refer to the
WAVE Bioreactor Instrumentation Manual.
DOOPT20 dissolved oxygen

monitor
Follow the instruction below to install the dissolved oxygen monitor.
Step Action
1 Connect the monitor to a grounded power outlet. The power connector is

2 Remove the DOOPT probe from its packaging. The probe can be left in its
protective tube until a calibration of the probe is needed.
3 Connect the DOOPT probe to the PROBE connector located on the front
panel of the monitor. Make sure that the ridge on the plug is lined up with
the notch on the connector. Push the plug in and twist to lock it in
position.
CO2MIX20 and CO2MIX20-R

CO2/air controllers
Follow the instruction below to install the CO2/air controllers.
Step Action
1 Connect the instrument to a grounded power outlet. The power

connector is located on the rear of the instrument.
2 Press the power switch to the on (I) position.
3 Set the external CO2 source between 0.7 and 1 bar (10 and 15 psig).
Connect the external CO2 source to the CO2 IN port on the front panel.
4 If installing CO2MIX20-R, adjust the CO2 pressure to 0.4 bar using the
regulator knob and the and the gauge located on the front panel of the
instrument. The regulator knob must be pulled to out to unlock.
5 If ambient air is to be mixed with the CO2 stream, leave the AIR IN port
unconnected. Otherwise, connect the desired air mixture to the AIR IN
port. External gas pressure must be regulated to between 0.1 and 0.2 bar
(1 and 3 psig).
6 Connect the MIX OUT port to the inlet filter on the Cellbag using the
tubing provided. The CO2 /air mixture is pumped out from this port.

3 Installation
3.6 Setting up external instrument modules
O2MIX20 and O2MIX20-R

O2/air controller
Follow the instruction below to install the O2/air controllers.
Step Action
1 Connect the instrument to a grounded power outlet. The power

connector is located on the rear of the instrument.
2 Press the power switch to the on position. Allow five minutes for the
instrument to warm up and the reading to stabilize.
3 Set the external O2 source between 0.7 and 1 bar (10 and 15 psig).
Connect the external O2 source to the O2 IN port on the front panel.
4 If installing O2MIX20-R, adjust the O2 pressure to 0.4 bar using the

regulator knob and the and the gauge located on the front panel of the
instrument. The regulator knob must be pulled to out to unlock.
5 If ambient air is to be mixed with the O2 stream, leave the AIR IN port
unconnected. Otherwise, connect the desired air mixture to the AIR IN
port. External gas pressure must be regulated to between 0.1 and 0.2 bar
(1 and 3 psig).
6 Connect the MIX OUT port to the inlet filter on the Cellbag using the
tubing provided. The O2 /air mixture is pumped out from this port.
PUMP20 peristaltic feed/

harvest pump
When used with a LOADCONT20 controller or a loadcell-equipped
WAVE Bioreactor 20/50, PUMP20 can be automatically switched on and off to deliver a
set feed rate or controlled harvest. When used with a shorting plug, PUMP20 can be
used as a variable speed standalone pump.
Follow any of the instructions below to install the peristaltic feed/harvest pump.
Use with LOADCONT20 or WAVE Bioreactor 20/50
Step Action
1 Connect the supplied DB9M-M cable to the connector on the rear of

PUMP20.
2 Connect the other end of the cable to the FEED PUMP or HARVEST PUMP
connector on the rear panel of the LOADCONT20 or
WAVE Bioreactor 20/50.

Installation 3
Transport 3.7
Standalone use
Step Action
1 Connect the power cord to the power connector located on the rear
panel of PUMP20 and to a grounded power outlet.
2 Connect the local shorting plug to the DB9 connector on the rear panel of
PUMP20. This will permit local operation.

Follow the instruction below to install the acid/base controller.
Step Action
1 Connect pH20 to a grounded power outlet. The power connector is

2 Connect the pH probe to the connector labeled SENSOR located on the

front panel of the pH20 unit.
3 Connect the acid/base PUMP20 units to pH20 using the DB9 connectors
labeled ACID and BASE located on the rear panel of pH20 and the
supplied cables.
3.7 Transport
Follow the instruction below to move WAVE Bioreactor 2/10 or WAVE Bioreactor 20/50.
Step Action
1 Disconnect the instrument from the power source.
2 Remove the Cellbag holder. The instrument should be moved without

liquid on top.
3 Lift the instrument by the lifting handles. The instrument should be moved
by two persons.
4 If installing the instrument in a new location, refer to the safety

instructions and following sections for installation procedures.
3.8 Spare parts and accessories

For correct up to date information on spare parts and accessories, visit
www.gelifesciences.com/wave or contact your local GE Healthcare representative.

3 Installation
3.8 Spare parts and accessories

Control systems 4
WAVE Bioreactor 2/10 control system 4.1
4 Control systems
This chapter describes basic operation of embedded control systems in the WAVE
Bioreactor base unit and external instrument modules. For some functions there are
different control options. For detailed instructions see relevant user documentation
listed in Section 8.2 Literature‚ on page 142.
4.1 WAVE Bioreactor 2/10 control system

WAVE Bioreactor 2/10 system is controlled by front panel controls, see Figure 4-1.
1 LCD display 7 AIR button
2 SETUP MENU ESC button 8 TEMP button
3 SETUP up and down arrow 9 PERF button

buttons
4 SETUP Enter button 10 ROCK, AIR, TEMP and PERF

LED indicators
5 ROCK, AIR and TEMP up and 11 ALM button

down arrow buttons
6 ROCK button 12 PAGE button
Figure 4-1. Front panel controls.

4 Control systems
4.1.1 LCD display main screen

For location of the LCD display, see Figure 4-1.
Item Function
#101 Communications address
rpm Rock speed (rocks per minute)
lpm Air flow (Liters per minute)
degC Temperature (ºC)
PV Process (actual) values

In this example 21 (rpm), 0.20 (lpm) and 32.0 (degC). These values
are not shown when the instrument is off)
SP Setpoint values
In this example 21/6º (rpm/rock angle), 0.20 (lpm) and 32.0 (degC)
ALARM! ALARM!
• A flashing ALARM! indicates a new alarm not yet
acknowledged.
• A steady ALARM! indicates an alarm that has been
acknowledged but not fixed.
• No ALARM! visible indicates that there is no current ALARM!
Note: If another screen is selected that screen is displayed.

Control systems 4
4.1.2 Rocking, aeration and

heating control
For location of the buttons, see Figure 4-1.
Rocking control
Step Action
1 Press ROCK button to start rocking. When rocking is on, the ROCK LED is
lit.
2 Press the ROCK up/down arrow buttons to change the rocking speed
(range 2 to 40 rpm). The selected rocking speed is shown on the setpoint
(SP) row.
3 Press ROCK button to stop rocking if rocking is on.
Note: If rocking is obstructed by a hand or other object, the safety switches stop the
rocking instantaneously. Reset power to resume operation.
Aeration control
.
Step Action
1 Press AIR button to start aeration. When aeration is on, the AIR LED is lit.
2 Press the AIR up/down arrow buttons to change the air rate (range 0.01
to 0.50 lpm). The selected air rate is shown on the setpoint (SP) row.
3 Press AIR button to stop aeration if aeration is on.
Note: The air pump automatically shuts down if the outlet pressure exceeds 3 inch H2O
for more than 10 seconds. This is to prevent over pressurizing the bag in the event
of clogging. The air flow resumes when the pressure returns to normal.
Heating control
.
Step Action
1 Press TEMP button to start heating. When heating is on, the TEMP LED is
lit.

4 Control systems
Step Action
2 Press the TEMP up/down arrow buttons to change the temperature

(range 10°C to 50°C). The selected temperature is shown on the setpoint
(SP) row.
3 Press TEMP button to stop heating if heating is on.
Note: The temperature system automatically shuts down if the unit is not rocking. This
is to prevent hotspots due to stagnant liquid motion. The temperature control
resumes once the unit is rocking again.
4.1.3 Perfusion control

Perfusion requires that an External instrument module for perfusion culturing,
PERFCONT2E, is installed. For location of the buttons, see Figure 4-1.
Step Action
1 Press PERF button to start the perfusion system. When perfusion is on,
the PERF LED is lit.
2 Perfusion parameters can be changed using menus accessible by

clicking the SETUP:MENU ESC button.
3 Press PERF button to stop the perfusion system.
Press PAGE button to view perfusion data and set points.
PAGE screen item Function
PERF wt Perfusion weight.

The current net weight (PV-prefix) and the weight setpoint
that is maintained by the harvest pump (SP-prefix) is
displayed in grams
shot The weight loss (or gain) in grams during each feed or
harvest operation.

Control systems 4
PAGE screen item Function
Σg Cumulative amount feed (prefix F) or harvest (prefix H). The

cumulative amounts can be cleared in the SETUP:CLEAR
MEMORY menu.
PV Process (actual) values

In this example 21 (rpm), 0.20 (lpm) and 32.0 (degC). These
values are not shown when the instrument is off
SP Setpoint values
In this example 21/6º (rpm/rock angle), 0.20 (lpm) and 32.0
(degC)
PERF ON / Indicates that perfusion is on/off.

PERF OFF
Note: Press PAGE button again to return to the Main menu.
4.1.4 Alarms
.
Step Action
1 A flashing ALARM! in the LCD display (see Fig 4-1) indicates a new alarm
not yet acknowledged.
2 Press ALM button to view alarm summary at any time. An example with a
temperature alarm (TMPDEV, temperature deviation) is shown.
3 When the ALM button is pressed the alarm status change to

acknowledged status and the ALARM! in the LCD display stops flashing.
4 To fix the Alarm, see Alarm messages‚ on page 120 for a list of alarm
messages and make appropriate corrective actions.
5 Press ALM button again to return to the previous view in the LCD display.

4 Control systems
4.1.5 Change operation

settings
Using SETUP buttons displayed parameter settings can be changed. For location of the
buttons, see Figure 4-1.
• Press SETUP MENU ESC button to enter the menu setup. There are 12 menus
available which are opened sequentially, see Table 4-1.
• Press SETUP up or down button to change a parameter value.
• Press SETUP Enter (↵) button to save a parameter value and open next menu.
• Press SETUP MENU ESC button while in a menu to return to the display main
screen.
Table 4-1. Operation parameters available using SETUP MENU ESC button.
LCD display Function
SET ANGLE
- Change the rocking angle
(range 2° to 9°)
SET PERFUSION
- Enable/disable perfusion option
SET TARE
- Tare the perfusion load cell
CLEAR MEMORY
- Clear stored cumulative feed/harvest amounts
SET WEIGHT SP
- Set the weight control setpoint

Control systems 4
LCD display Function
SET PERF RATE

- Set the desired daily perfusion rate
SET FEED SHOT

- Set the desired feed shot in mL
CALIB TEMPERATURE
- Calibrate the temperature sensor
STOP POSITION
- Set the sample stop position (default 9º)
UNIT ADDRESS
- Set the Unit Address
(range 101 to 110)
AUTOSTART
-If the AUTOSTART option =1 controls are
automatically restored to the ON state on power
up after shutdown or power lost.
CAL LEVEL
- Set the calibration level. This allows adjusting the
level.
LEVEL@deg/10= 1 means 1 divide by 10 which
will give an 0.1 offset.

4 Control systems

WAVE Bioreactor 20/50 system is controlled by a Touch screen. The Touch screen main
screen functions available varies depending on the current system configuration, see
Figure 4-2 for an example.
• The black boxes are function buttons. Pressing one of these will bring up the
corresponding detail screen.
• Boxes display to values of various parameters such as speed, temperature, and

airflow.
• If alarm conditions exist, then the ALARM button will be displayed at the bottom of
the screen. If the ALARM is new and has not been acknowledged, then the ALARM
button will be flashing.
1 2 3 4
10
11
5 6 7 8 9
1 Rock speed control 7 Alarm indicator (ALARM)

(ROCKER)
2 Actual rocking speed (rpm) 8 Aeration control (AIR)
3 Heating control (HEATER) 9 Actual airflow (lpm)
4 Actual temperature of 10 O2MIX button for control of

Cellbag (ºC) O2 concentration
5 Weight of Cellbag control 11 System setup and other

(WGHT) functions (MENU)
6 Actual weight of Cellbag (Kg)
Figure 4-2. Main screen example showing a Dual option EHTD system. Available functions varies
depending on system configuration

Control systems 4
Each of the function has a screen which opens by pressing respective button. For details
of control system functions, refer to the WAVE Bioreactor 20/50 Operator Manual
4.2.1 General screen functions

Functions available in most or all of different functional screens.
Function Description
TREND Press the TREND button to get a graph for the parameter
over a 10 minute period.
ALARM See Section 4.2.8.
BACK Press the BACK button to return from current screen to the
main screen.
4.2.2 Rocking speed control

.
Step Action
1 In the main screen (Fig 4-2), press ROCKER.
2 The ROCKER screen opens.
Current rocking speed is shown in the rpm box.
3 Set the rocking speed set point by pressing the SP box. Enter desired set
point (2 to 40 rpm) in the data entry keypad that opens and press the ↵
key.
4 Set the rocking angle by pressing the angle box. Enter desired set point (2
to 12 degrees) in the data entry keypad that opens and press the ↵ key.
5 Press ROCK to start rocking and check that the unit rocks properly.
6 Press STOP (appears in the same position as ROCK) to stop rocking.

4 Control systems
4.2.3 Temperature control

Note: The heating switches off if:
- Temperature sensor fails
- Heater is unplugged
- Heater overheats (> 60°C)
- Unit not rocking (this may be overridden in SETUP)
Note: When using a Dual system with a large single bag (20L, 22L or 50L) use Single
setup operation..
Step Action
1 In the main screen (Fig 4-2), press HEATER.
2 The HEATER screen opens (the figure shows a Dual setup screen with two
sets of commands, LEFT and RIGHT, see Section 4.2.9 to change setup).
Current Cellbag temperature is shown in the box labeled °C.
3 Set the temperature set point by pressing the SP box. Enter desired set
point (0°C to 45°C) in the data entry keypad that opens and press the ↵
key.
4 Press HEAT to start heating.

The power output of the heater is represented in the barograph
(Rectangular box to the left of °C and SP boxes in the LEFT part of the
screen and right of °C and SP boxes in the RIGHT part of the screen)
5 Press STOP (appears in the same position as HEAT) to stop heating.
4.2.4 Weight control

Note: Weight control requires BASE20/50EHT-L system version.
Step Action
1 In the main screen (Fig 4-2), press WGHT.

Control systems 4
Step Action
2 The WEIGTH screen opens.
The current net weight in kg is shown in the Kg net box
3 Set the net weight set point by pressing the SP box. Enter desired set
point (0.2 to 25 kg) in the data entry keypad that opens and press the ↵
key.
4 Start the feed pump by pressing the ON button below FEED.

Note: The FEED pump does not turn on if the net weight is higher than the
high alarm limit.
• The button changes to OFF and the indicator Feeding appears.

• The feed pump runs until the net weight is above the setpoint.
• Press OFF to disable the FEED controller.
5 Start the harvest pump by pressing the ON button below HARV.

Note: The harvest pump (HARV) does not turn on if the net weight is less
than the low alarm limit.
• The button changes to OFF and the indicator Harvesting appears.

• The harvest pump runs until the net weight is below the setpoint.
• Press OFF to disable the FEED controller.
6 To set net weight to zero, press TARE. The TARE NOW? screen opens.
• Press TARE NOW to set the net weight to zero. This cannot be
undone.
• Press CANCEL to return to the WEIGTH screen.

4 Control systems
Step Action
7 To use perfusion control press PERFU. The PERFUSION screen opens.
The left side of the screen shows the feed information:

• Current status of feed pump: F_PUMPOFF or F_PUMPON
• Amount feed added (shot)/to be added in this shot (gm).
• Cumulative feed (L) is shown in the Σ box.
The left side of the screen shows the harvest information:
• Current status of harvest pump: H_PUMPOFF or H_PUMPON
• Amount harvest removed (shot) /to be removed (gm).
• Cumulative harvest (L) is shown in the Σ box.
The current net weight in kg is shown in the Kg box.
8 Press the RESET button to zero out both the feed and harvest cumulative
amounts.
9 Press DETAIL button.
10 The PERFUSION PARAMETERS screen opens.
• Set the total feed by day (L/d) pressing the FEED box. Enter desired
set point in the data entry keypad that opens and press the ↵ key.
• Set the feed volume per shot by pressing the Shot ml box. Enter
desired set point in the data entry keypad that opens and press the ↵
key.
• The period in minutes between each shot is calculated by values set
for total feed by day and feed volume per shot.

Control systems 4
Step Action
11 Set the net weight set point by pressing the SP box. Enter desired set
point (0.2 to 25 kg) in the data entry keypad that opens and press the ↵
key.
12 Press ON to start the feed and harvest control.

• The feed pump starts and the harvest pump is activated when the
weight exceeds the setpoint.
• The ON button changes to OFF. Press OFF to disable the perfusion
controller.
4.2.5 Aeration control

.
Step Action
1 In the main screen (Fig 4-2), press AIR.
2 The AIRPUMP screen opens (the figure shows a Dual setup screen)
Current air flow is shown in the lpm box.
3 Set the aeration set point by pressing the desired SP box. Enter desired
set point (0 to 0.5 Lpm) in the data entry keypad that opens and press the
↵ key.
4 Press ON (left or right in Dual setup) to start the air pump.
5 Press STOP (appears in the same position as ON) to stop the air pump.

4 Control systems
4.2.6 CO2 concentration /

aeration control
Note: CO2 concentration control requires BASE20/50EHTD-CO2 instrument
configuration.
Air flow is required for the CO2 controller to work properly. For convenience, airflow
controls are provided on the right side of the CO2 screen. For information of controlling
the airflow, see Section 4.2.7
.
Step Action
1 In the main screen (Fig 4-2), press CO2.
2 The CO2 / AIRPUMP screen opens)
Current CO2 concentration is shown in the % CO2 box.
3 Set the CO2 concentration set point by pressing the desired SP box. Enter
desired set point (0% to 15%) in the data entry keypad that opens and
press the ↵ key.
4 Press ON to start the air pump.
Note: CO2 gas must be connected to the right side of the unit. The pressure must be
regulated to between 10 and 15 psig and capable of delivering at least 0.2 Lpm.
4.2.7 O2 concentration /
aeration control
Note: O2 concentration control requires BASE20/50EHTD-O2 instrument configuration).
Air flow is required for the O2 controller to work properly. For convenience, airflow
controls are provided on the right side of the O2 screen. For information of controlling
the airflow, see Section 4.2.7.

Control systems 4
Step Action
1 In the main screen (Fig 4-2), press O2.
2 The O2 / AIRPUMP screen opens)
Current O2 concentration is shown in the % O2 box.
3 Set the O2 concentration set point by pressing the desired SP box. Enter
desired set point (21% to 50%) in the data entry keypad that opens and
press the ↵ key.
4 Press ON to start the air pump.
Note: O2 gas must be connected to the right side of the unit. The pressure must be
regulated to between 10 and 15 psig and capable of delivering at least 0.2 Lpm.
4.2.8 Alarms
An new alarm is indicated by:
• A flashing ALARM button appears on the bottom of the displayed screen.
• The screen color changes to red. This indicates that the alarm has not been
acknowledged.
• An audible beeper sounds. The beeper can be disabled, see WAVE Bioreactor 20/
50 Operator Manual.
To handle an alarm
All alarms have a specified code and are time-stamped to facilitate troubleshooting
(Alarm messages‚ on page 126).
Step Action
1 In the main screen (Fig 4-2), press the ALARM button.

4 Control systems
Step Action
2 The ALARMS screen opens. This action also acknowledges the alarm and
returns the screen color to green.
3 On the ALARMS screen each alarm is recorded. Each alarm message has:
• The date and time the alarm occurred or when it was cleared.
• Alarm status (Alm = not cleared, OK = cleared).
• The alarm code number, and short description, see Alarm messages‚
on page 126 (underscore between).
4 Press Up or Down buttons to scroll through the alarm history list.
5 To access a help screen for alarms, press desired alarm to highlight it.
and press HELP.
6 To get a list of currently active alarms, press the Active Alarms button.
7 Solve the alarm issue.
8 Once an alarm has been acknowledged, the ALARM button at the bottom
of the main screen stops flashing. It remains visible as long as any alarm
condition exists.
9 To view the alarm history and active alarm list, press the ALARM button
in the main screen (if visible) or accessed press MENU and then ALARM in
the MENU screen.
10 The alarm history is stored in the Touch screen. The alarm history is
maintained even if power is lost. The last 1000 alarms are retained. If the
Touch screen is interchanged between base units, the alarm history is
lost.

Control systems 4
Step Action
11 Autoacknowlegde:
Normally the user must acknowledge any new alarm by pressing the
ALARM button to view the alarm history screen. If the AUTOACK feature
is set to ON on the SETUP:OPTIONS screen then all alarms are
automatically acknowledged. No warnings are generated but the alarm
is still logged.
12 Alarm contacts
If any alarm condition exists, this triggers the UNIT ALARM. This sets the
external alarm contact. The UNIT ALARM goes OFF only if no alarm
conditions are present.

settings
For detailed instructions see the WAVE Bioreactor System 20/50EHT Operator Manual.
Step Action
1 In the main screen (Fig 4-2), press MENU.
2 The MENU screen opens.
The MENU screen shows system information.
3 Press the ALARM to open the ALARMS screen described in Section 4.2.8.
4 Press the SETUP button to edit instrument set up.

4 Control systems
4.3 WAVEPOD control system
Step Action
5 The Access SETUP screen opens.
Click on the Enter Password box and enter password (default password
is 2050) followed by the ↵ key.
6 The SETUP screen opens.
• The list of setup buttons displayed depends on the configuration of

the instrument and only the relevant set up buttons are displayed.
• All selections are retained in non-volatile memory and are be
preserved on power off.
• Press desired button to change setup for corresponding instrument
settings, see WAVE Bioreactor System 20/50EHT Operator Manual for
details.

The WAVEPOD external instrument module combines a number of instruments for use
with WAVE Bioreactor system. It can provide dissolved oxygen measurement and
control; pH measurement and control; aeration control and O2 and/or CO2 gas blending.
A WAVEPOD unit is controlled by a Touch screen. In the Touch screen there are several
different control screens used in the control of WAVEPOD. The Main (Overview) screen is
shown in Figure 4-3. Note that only functions available in the current system setup is
displayed.

Control systems 4
WAVEPOD control system 4.3
2
1
9
3 13
4 10 14
5
11 15
6
12 16
7 17
8 19
18
1 Connected Bioreactor base 11 Measured dissolved oxygen

unit (%sat)
2 Time and date 12 DO OPT button for control of

dissolved oxygen
3 Actual rocking speed (rpm) 13 Measured O2 (%O2)
4 Actual aeration value (lpm) 14 O2MIX button for control of

O2 concentration
5 Actual temperature value 15 Measured CO2 (%CO2)

(°C)
6 Perfusion option only: 16 CO2MIX button for control

Actual net weight value (Kg) of CO2 concentration
7 Connected Bioreactor base 17 Aeration indicator

unit (2050EHT) button (green on/red off)
8 MENU button for operation 18 ALARMS button, if present it

settings indicates one or more
alarms
9 Measured pH
10 pH OPT button for pH 19 HELP button for information

control of various parameters.
Figure 4-3. WAVEPOD main screen. Green/red bars indicate that actual values are within/outside
setpoint value limits.

4 Control systems
4.3.1 General buttons

Functions available in most or all of different functional screens.
Function Description
TREND Press the TREND button to get a graph for the parameter
over a 10 minute period. WAVEPOD does not maintain long-
term trends.
A typical TREND screen is shown below.
ALARMS See Section 4.3.8.
BACK Press the BACK button to return to the main screen from
current screen.

Control systems 4
4.3.2 pH control
..
Step Action
1 Press the pH OPT button in the main screen (Fig 4-3).
2 The pH CONTROL screen opens.
The pH CONTROL screen provides:

• Actual pH (pH) display box
• Setpoint pH (SP) display box
• pH control button (CONTROL ON/OFF)
• ACID BASE/CO2 BASE control option selection
• TREND, ALARMS and BACK buttons.
3 Click the SP display box to change pH setpoint value.

• A data entry keypad opens.
• Enter a new setpoint (pH range 5 to 9)
• Press the ENT key to accept the new setpoint, CLR to clear the
display and ESC to abort data entry.
4 Press CONTROL ON/OFF button to start pH control.

• The button changes to green and displays ON.
• Pressing it again disables pH control and the button changes to red
and displays OFF.

4 Control systems
Step Action
5 Press ACID BASE/CO2 BASE control option selection to select mode for
pH control.
• ACID BASE option:
pH control by Acid/Base pumps.
• CO2 BASE option:
pH control by CO2 concentration and a Base pump.
4.3.3 Dissolved oxygen control

.
Step Action
1 Press the DO button in the main screen (Fig 4-3).
2 The DO OPTICAL screen opens.
The DO OPTICAL screen provides:

• DO Probe bar graph (left of actual DO concentration field (DO %sat))
• Actual DO concentration display box (DO %sat).
• Setpoint DO (SP) display box showing the setpoint value of dissolved
oxygen.
• T/COMP display box (ºC)
• DETAIL button
• CALIB button
• SCAN display box (s)
• CONTROL ON/OFF button
• RPM/O2% control option selection

Control systems 4
Step Action
3 DO Probe bar graph indicates the condition of the DO probe. When the
bar graph is green, the probe is functional. When the green bar reaches
the bottom, the graph turns red indicating the probe should be replaced.
4 Press the SP display box.

• Enter a new setpoint (range 0 to 100 +/- 0.5%sat)
5 Press T/COMP display box.

• Enter a new setpoint temperature of the DO probe. This is used for
temperature compensation.
6 Press DETAIL to open a pop-up window displaying various probe

parameters, such amplitude and phase of the signal. This display is
useful for troubleshooting.
7 Press CALIB to calibrate the DO probe.

• A probe adjustment screen opens.
• Follow instructions on the screen.
• Press CLOSE when calibration is done.
Note: See also Section 6.4.
8 Press SCAN display box to set the time between successive DO

readings.
• Enter a new setpoint time of the DO probe (range 1 to 99 seconds).
9 Press CONTROL ON/OFF button to start dissolved oxygen (DO) control.

• Pressing it again disables DO control and the button changes to red
and displays OFF.

4 Control systems
Step Action
10 RPM/O2% control option selection

• Rocking rate (RPM) option:
Dissolved oxygen control by change of the rocking rate.
• O2 concentration (O2%) option:
Dissolved oxygen control by change of O2 concentration.
4.3.4 Aeration control

See Section 4.3.5 or Section 4.3.6.
4.3.5 O2 concentration control

.
Step Action
1 Press the O2MIX button in the main screen (Fig 4-3).
2 The O2MIX CONTROL screen opens. Air flow is required for the O2
controller to work properly. Airflow controls are provided on the right side
of the O2MIX CONTROL screen (REMOTE BASE)
The O2MIX CONTROL screen provides:

• Actual O2 concentration display box (O2 %).
• Setpoint O2 (SP) display box showing the setpoint value of O2
concentration.
• O2 supply pressure indicator (Green circle indicates O2
concentration within setpoint values).
• Actual airflow (REMOTE BASE MIX OUT) display box (lpm)
• Setpoint airflow (REMOTE BASE MIX OUT) display box (SP)

Control systems 4
Step Action
3 Press the setpoint O2 SP display box.

• Enter a new setpoint (range 0 to 50% O2, Control range is 21 to 40%
+/- 0.6%, 41 to 50% +/- 1.0%).
4 Press CONTROL ON/OFF button to start O2 control.

and displays OFF.
5 Optional: The O2 setpoint can be set remotely by another controller (RSP,

Remote Set Point). When the option is active, a flashing message, RSP
from DO, is displayed.
6 Press the Aeration (MIX OUT) setpoint O2 concentration SP display box.

• Enter a new setpoint (range 0 to 0.5 Lpm).
7 Press AIR ON/OFF to start aeration control.

• Pressing it again disables aeration control and the button changes to
red and displays OFF.

4 Control systems
4.3.6 CO2 concentration

control
.
Step Action
1 Press the CO2MIX button in the main screen (Fig 4-3).
2 The CO2MIX CONTROL screen opens. Air flow is required for the CO2
controller to work properly. Airflow controls are provided on the right side
of the CO2MIX CONTROL screen (REMOTE BASE)
The CO2MIX CONTROL screen provides:

• Actual CO2 concentration display box (CO2 %).
• Setpoint CO2 (SP) display box showing the setpoint value of CO2
concentration.
• CO2 supply pressure indicator (Green circle indicates CO2
concentration within setpoint values).
• Actual airflow (REMOTE BASE MIX OUT) display box (lpm)
• Setpoint airflow (REMOTE BASE MIX OUT) display box (SP)
3 Press the setpoint CO2 SP display box.

• Enter a new setpoint (range 0 to 15% CO2).
4 Press CONTROL ON/OFF button to start CO2 control.

and displays OFF.

Control systems 4
Step Action
5 Optional:
The CO2 setpoint can be set remotely by another controller (RSP, Remote
Set Point). When the option is active, a flashing message, RSP from pH, is
displayed.
6 Press the Aeration (MIX OUT) setpoint CO2 concentration SP display box.
• Enter a new setpoint (range 0 to 1.00 Lpm).
7 Press AIR ON/OFF to start aeration control.

• Pressing it again disables aeration control and the button changes to
red and displays OFF.
4.3.7 Bioreactor base unit

control
A WAVE Bioreactor 20/50 base unit can be remotely controlled from WAVEPOD. If the
Bioreactor base unit is connected and operating, overview data are displayed on the
left side of the main screen (Fig 4-3).
Press the Bioreactor base unit (2050EHT) button. See the WAVEPOD Operator Manual for
instructions.
4.3.8 Alarms
All alarms are time-stamped and have a short description to facilitate troubleshooting,
see (Alarm messages‚ on page 131) for a list of alarms.
The alarm beeper sounds every time a new alarm occurs. The beeper can be disabled
(ALM BEEPER OFF) from the SETUP OPTIONS screen.
Step Action
1 A red flashing ALARMS button appears on the bottom of the displayed

screen to indicate a new Alarm. In addition an audible beeper sounds (to
disable the beeper, see Section 4.3.9.
2 Press the ALARMS button to access the alarm history screen. This action
also acknowledges the alarm and makes the ALARMS button turn yellow
and stop flashing. The ALARMS button remains visible until all alarms
have cleared.

4 Control systems
Step Action
3 The ALARM LOG screen opens.
Each alarm entry has the date and time, alarm number, description and
status (red=current unacknowledged alarms, blue=reviewed alarms,
green=cleared alarms).
The alarm history is retained on power failure or shutdown. The last 1000
alarms are retained.
• Use the up and down scroll arrows to page through the alarm log.
• The alarm history can be permanently cleared from the SETUP menu
(MENU:SETUP: CLEAR ALARM LOG).
• To review an alarm, press the alarm message. This brings up a pop-
up window with more information about the alarm. The alarm
message turns blue indicating that alarm has been viewed.
4 The ALARM LOG can be reviewed at any time by pressing the ALARMS
button from the main screen (Fig 4-3) or by pressing the REVIEW ALARMS
button in the SYSTEM INFO screen.

Control systems 4

settings
Step Action
1 Press the MENU button in the main screen (Fig 4-3).
2 The SYSTEM INFO screen opens.
The SYSTEM INFO screen provides:

• WAVEPOD serial number (POD serial #)
• PLC version number (PLC ver)
• Touch screen program version (TPanel ver)
• Compile date (Compiled)
• ALARM STATUS, for Bay 1 and 3 (B1&3), Bay 2 and 4 (B2&4) and
Bioreactor base unit (BASE).
Status:
• Green = No current Alarm (ALM)/acknowledged alarm (ACK)/
communication (COM)
• Red = Current Alarm (ALM)/unacknowledged alarm (ACK)/

no communication (COM)
• REVIEW ALARMS button provides the current alarm status, see

Section 4.3.8.
• SETUP button, see step 3.

4 Control systems
Step Action
3 Press SETUP in the SYSTEM INFO screen.

A password entry screen OPENS. Password prevents unauthorized
personnel from accessing SETUP parameters. The default password is
2050.
• Press the XXXX button and enter the password and press ENT.
• Press the PROCEED button which is displayed.
4 The SETUP screen opens. Options available in the SETUP screen depends
on the specific system configuration. For detailed description of setting
parameters, see the WAVEPOD Operator Manual.
5 Click OPTIONS button to set Bioreactor base unit type, autostart after
power failure (on/off), autoacknowledge of Alarms (on/off) and Alarm
beeper (on/off).
6 Click SET CLK button to set instrument time.
7 Click CLEAR ALARM LOG button to clear the stored alarm log. The alarm
log cannot be retrieved.

Control systems 4
Step Action
8 Click DO OPT button.
In the DO CONTROL SETUP screen it is possible to set:

• Deviation alarm (DEV alarm), ON/OFF
• Deviation limits (DEV limit), range 0% to 10%
• Dead band range for DO (DBAND), range 0% to 10%
• Cycle time (CTIME), that is the time to allow the acid/base or CO2
change to result in a stable pH reading, range 10 to 999 seconds.
• Setpoint values for rocking speed (rpm) and O2 (CONTROL SCHEME).
9 Click pH button.
In the pH CONTROL SETUP screen it is possible to set:

• pH deviation alarm (DEV alarm), ON/OFF
• Deviation limits (DEV limit) range 0.0 to 1.0 pH unit
• Dead band range for pH (DBAND) range 0.0 to 1.0 pH unit
• Cycle time (CTIME), that is the time to allow the acid/base or CO2
change to result in a stable pH reading, range 10 to 300 seconds.
• Setpoint values for acid, base and CO2 (CONTROL SCHEME).

4 Control systems
Step Action
10 Click O2 MIX button.
In the O2MIX CONTROL SETUP screen it is possible to:

• Set O2 concentration deviation alarm (DEV alarm), ON/OFF.
• Set O2 deviation alarm deadband (deviation limits, DEV limit) range
0.0% to 5.0%.
• View actual O2 concentration (O2 PV).
• Set O2 OFFSET, small adjustments of O2 concentration settings can
be made by changing the offset.
• Set airflow deviation alarm (AIRFLOW DEV alarm), ON/OFF. Deviation
limit is fixed at 0.02 Lpm.
• Set SCALING (ml/min): Small airflow adjustments can be made by
changing the scaling.

Control systems 4
Step Action
11 Click CO2MIX button.
In the CO2MIX CONTROL SETUP screen it is possible to:

• Set CO2 concentration deviation alarm (DEV alarm), ON/OFF.
• Set CO2 concentration deviation alarm deadband (DEV limit), +/-
CO2%, range 0.0% to 5.0%.
• View actual CO2 concentration (CO2 PV).
• Set CO2 OFFSET, small adjustments of CO2 concentration settings
can be made by changing the offset.
• Air flow deviation alarm (AIRFLOW DEV alarm), ON/OFF. Deviation
limit is fixed at 0.02 Lpm.
• Set SCALING (ml/min): Small airflow adjustments can be made by
changing the scaling.

4 Control systems
4.3.10 Help
Step Action
1 Press the HELP button in the main screen (Fig 4-3).
2 WAVEPOD HELP screen opens.
3 For getting information of different topics click the corresponding button,

see the WAVEPOD Operator Manual for details.
4 Click BACK button to return to the main screen.

Control systems 4
External instrument module control systems 4.4
4.4 External instrument module control systems
4.4.1 Common Process

controllers
Several external instrument modules for the WAVE Bioreactor 2/10 and WAVE
Bioreactor 20/50 are available. Process controllers of two sizes are integrated in
external instrument modules in which actual and setpoint values can be viewed and
setpoint values can be changed, see Figure 4-4 and Table 4-2.
Controller item Function
• 1, 2, 3, 4 (Large controller) Output indicators light when corresponding

output is activated.
• 1, 2 (Small controller)
% Auto/manual indicator lights when corresponding

output is activated
Upper display: red digits Shows process value or parameter value
Lower display: green digits Shows setpoint or parameter name.
Press to display through menu items or

TAdvance button parameters
Up arrow button Press to increase setpoint or parameter
Down arrow button Press to decrease setpoint or parameter
∞ Home button Press to return to normal operation
Figure 4-4. Large Process controller (left) and small Process controller (right).

4 Control systems
Table 4-2. Large and small Process controller instrument modules.
Large Process controller module Small Process controller module
O2MIX20 O2/air controller DOOPT20 dissolved oxygen monitor
O2MIX20-R O2/air controller O2MIX20 O2/air controller
CO2MIX20 CO2/air controller CO2MIX20 CO2/air controller
CO2MIX20-R CO2/air controller
4.4.2 Control of DOOPT20

dissolved oxygen monitor
See Section 1.8 for location of buttons and Section 4.4.1 for description of Process
controller module.
NOTICE
Sensitive DO probe. Do not touch the tip of the probe with your finger or
any object. Do not attempt to clean or dry the tip with a cloth. Simply
shake dry
Step Action
1 Connect the probe to the connector on the front panel. Make sure that
the ridge on the plug is lined up with the notch on the connector. Push the
plug in and twist to lock it in position, do not force the plug.
2 Turn the power switch on (I). The power switch is located on the back of
the instrument module.
3 The displays should light up and a version number is displayed on the

LCD display.
4 Within a few seconds, the LCD display shows T= temperature

compensation value and A= amplitude.
Note: When the A -value is less than 5000, the DO Probe has
approximately 3 to 4 weeks of use left. Replace the probe
immediately if A -value is under 2000.
5 After a period of 15 seconds, the display indicates the temperature in °C

and P = phase angle of the DO probe. The phase angle and amplitude
displays alternate every 15 seconds.

Control systems 4
Step Action
6 The Process control module show the dissolved oxygen value (red).
Note: Please allow the unit to scan the probe, be patient.
7 Calibrate the DO probe before use, see Section 6.4 for instructions
8 Insert the DO probe in Cellbag.

Notes: The DO probe can to be removed and reinserted repeatedly without
any danger of contamination.
• Locate the Oxywell2 fitting on the Cellbag.

• Remove the luer cap from the fitting.
• Fill the Oxywells with distilled water using the fill kit provided with the
DO probe. instructions are included with the probe. This is critical to
provide a response time (t90) of about 3 to 5 minutes.
• Insert the DO probe carefully into the Oxywell2.
• Secure the DO probe by tightening the luer connector.
9 For optimum accuracy readjust the probe after insertion into the bag
(possible if the Cellbag has not been inoculated). With aeration and
rocking this should correspond to the 100% saturation point.
10 Let the Cellbag equilibrate so that it is at the correct temperature and at

100% air saturation.
11 Set the manual temperature compensation to the temperature of the

Cellbag and re calibrate the 100% point.
4.4.3 Control of CO2MIX20 CO2/

air controller
controller module.
Use as aeration pump only
Step Action
2 Press the PUMP ON switch to start the air pump.

4 Control systems
Step Action
3 Set the air flow setpoint by the up/down arrow buttons on the airflow
controller. The setpoint is shown in green. The actual airflow is shown in
the red. Control range is 0 to 0.5 liters/minute.
4 Air is drawn from the AIR IN inlet connector located on the front of the
instrument. Any special gas mixture can be connected here in place of
room air. External gas pressure must be regulated to between 0.1 and 0.2
bar (1 to 3 psig).
Use as CO2/air mix controller
Step Action
4 Press the CO2 ON switch to switch on the CO2 controller. The switch lights
up indicating that CO2 control is active.
5 Select the %CO2 desired by adjusting the setpoint on the Process

controller. The setpoint is displayed in green. The actual CO2 reading is
displayed in red.
The setpoint can be changed by the up/down arrow buttons.
High pressure alarm and

shutoff
If the pressure at the AIR OUT port exceeds 4 inches H2O due to blockage or obstruction
of the air flow to the Cellbag, the red HIGH PRESSURE LED starts blinking. If the
overpressure condition continues for longer than 1 to 2 minutes, the integral air flow
valve shuts down preventing the pressure from rising higher. When the overpressure
condition clears, the airpump resumes operation.
4.4.4 Control of CO2MIX20-R

CO2/air controller
controller module.

Control systems 4
Step Action
3 Air flow can be regulated by the valve on the rotameter.
bar (1 to 3 psig).
Use as CO2/air mix controller
Step Action
3 Press the CO2 button to switch on the CO2 controller. The button lights up
indicating that CO2 control is active.
4 Select the %CO2 desired by adjusting the setpoint on the Process

controller. The setpoint is displayed in green. The actual CO2 reading is
displayed in red.
4.4.5 Control of O2MIX20 O2/air

controller
controller module.
Step Action

4 Control systems
Step Action
bar (1 to 3 psig).
Use as O2/air mix controller
Step Action
4 Press the O2 ON switch to switch on the O2 controller. The switch lights

up indicating that O2 control is active.
5 Select the %O2 desired by adjusting the setpoint on the Process

controller. The setpoint is displayed in green. The actual O2 reading is
displayed in red.
High pressure alarm and

shutoff
If the pressure at the AIR OUT port exceeds 3 inches H2O due to blockage or obstruction
of the air flow to the Cellbag, the red HIGH PRESSURE LED starts blinking. If the
overpressure condition continues for longer than 1 to 2 minutes, the integral air flow
valve shuts down preventing the pressure from rising higher. When the overpressure
condition clears, the airpump resumes operation.

Control systems 4
4.4.6 Control of O2MIX20-R O2/

air controller
controller module.
Step Action
3 Air flow can be regulated by the valve on the rotameter.

bar (1 to 3 psig).
Use as O2/air mix controller
Step Action
3 Press the O2 button to switch on the O2 controller. The button lights up

indicating that O2 control is active.
4 Select the %O2 desired by adjusting the setpoint on the Process

controller. The setpoint is displayed in green. The actual O2 reading is
displayed in red.

4 Control systems
4.4.7 Control of PUMP20

peristaltic feed/harvest pump
See Section 1.8 for location of buttons.
Step Action
1 Feed the tubing so that the inlet side is located at the lower clip and the
outlet in the upper clip. Rotate the pumphead and ensure the tubing is
securely held with no slack. The roller tension should be adjusted. Adjust
the clips so that the tubing is not pulled into the pump when the head
rotates.
Note: Pump always runs in a clockwise direction.
2 Press the PUMP ON switch to turn power on. The PUMP ON button is lit.
• If connected to a LOADCONT20 or BASE20/50EHT, the pump does not
run unless this device sends a REMOTE ON signal through the DB9
connecting cable. The pump runs at the speed set on the panel digital
indicator.
• If the shorting plug is installed, then the pump runs at the speed set
on the panel digital indicator.
3 Press the RUN button to run the pump at full speed as long as the button
is held down. PUMP ON switch indicator light must be on (green). This
function is useful for priming tubing.
4 The RUN button lights up whenever the pump is running.
5 The pump speed can be set 10% to 100% by pushing the +/- buttons on
the digital speed control.
4.4.8 Control of pH20 acid/

base controller
controller module.
General recommendations
For stable pH control it is necessary to cycle the addition pumps so that the acid or
base has sufficient time to mix properly. The pH20 controller has a cycle time of a
preset duration (default= 30 seconds). Part of this cycle is the on time when the acid
or base pump is energized. For the remainder of the cycle the pumps are off and
the controller waits until the cycle time is complete.

Control systems 4
• If, at the end of the cycle, the pH is still not at the setpoint, the controller initiates
another add/wait cycle.
• If the pH is at the setpoint, the controller does nothing. A small deadband around
the setpoint (default = 0.1 pH) is used to prevent excessive pump chatter.
The cycle time can be adjusted if the system requires more mixing time. The
amount of acid or base added per cycle is proportional to the difference between
the setpoint and the actual pH. The Gain constant adjusts the sensitivity. This Gain
can be decreased if insufficient acid or base is being added per cycle. It can also be
increased if very concentrated acid or base is being used and significant overshoot
is observed.
Connecting the base pump
Step Action
the instrument module. Make sure that the BASE ENBL button is off.
2 A peristaltic base pump such as the WAVE PUMP20 can be controlled by

the pH20 controller. If using PUMP20 units, plug the PUMP20 for the base
pump to the Controller using the supplied cables.
3 Make sure that the pump direction is correct and turn all local switches
on. Route the base tubing through the pump and connect to the Cellbag
container. Open all valves.
4 Set the desired pH setpoint on the controller and press the BASE ENBL
button to activate base control.
5 The base pump turns on whenever the pH drops 0.1 unit below the pH
setpoint.
Connecting the acid pump
Step Action
the instrument module. Make sure that the ACID ENBL button is off.
2 A peristaltic base pump such as the WAVE PUMP20 can be controlled by

the pH20 controller. If using PUMP20 units, plug the PUMP20 for the base
pump to the Controller using the supplied cables.
3 Make sure that the pump direction is correct and turn all local switches
on. Route the base tubing through the pump and connect to the Cellbag
container. Open all valves.

4 Control systems
Step Action
4 Set the desired pH setpoint on the controller and press the ACID ENBL
button to activate base control.
5 The acid pump turns on whenever the pH exceeds 0.1 unit below the pH
setpoint.
Setting the pH setpoint

The pH setpoint determines the pH that should be maintained in the Bioreactor. To
change the setpoint:
Step Action
1 The green digits display shows the current setpoint.
2 Press the up/down arrow buttons to change the setpoint. The controller
responds once the button is released.
Changing the controller gain

The controller gain changes the percentage of on time during each cycle.
Increasing the Gain increases the amount of time the pump stays on at the same
error (setpoint –actual pH). Acid and base gain can be set independently. pB1 is the
base gain and pB2 is the acid gain (default=5.0).
To change the gains:
Step Action
1 Press the Process controller Advance button Tto access the menu.
Press theT button repeatedly until the desired parameter (pB1 or pB2)
is displayed on the green LED. Increase the pB values to reduce the
amount of acid/base added per shot.
• Decrease the pB values to increase the amount of acid/base per
shot.
2 Press the up/down arrow buttons to change the displayed gain. Press the
α button to return to normal operation.
Changing the cycle time

The cycle time gives the acid or base liquid time to mix. If the cycle time is too short
then additional acid or base may be added before the reading has stabilized. If the
cycle time is too long then the response will be sluggish. Acid and base cycle times

Control systems 4
can be set independently. Ct1 is the base cycle time and Ct2 is the acid cycle time.
The cycle time is in seconds (default=30.0 seconds).
To change the cycle time:
Step Action
1 Press the Process controller Advance button Tto access the menu.
Press the T button repeatedly until the desired parameter (Ct1 or Ct2)
is displayed on the green LED.
2 Press the up/down arrow buttons to change the displayed cycle time.
Press the ∞ button to return to normal operation.
Changing the deadband

A deadband is required around the setpoint to prevent the pumps from cycling too
frequently. The acid and base dead bands can be set independently. dB1 is the
base cycle time and dB2 is the acid cycle time. The deadband is in pH units
(default=0.1). To change the deadbands:
Step Action
1 Press the Process controller Advance button T to access the menu.

Press the T button repeatedly until the desired parameter (dB1 or dB2)
is displayed on the green LED.
2 Press the up/down arrow buttons to change the displayed deadband.

Press the ∞ button to return to normal operation.
Using user supplied pumps

Any pump capable of being run by a remote output can be interfaced to the pH20
acid/base controller.
Pins 1 and 9 on the DB9 connector are the emitter and collector of an open
collector output respectably. Whenever the controller needs a pump to run, the
output is turned on. The output load current must not exceed 100mA.
User supplied pumps can be also connected to the pH20 acid/base external
instrument module by using the optional Power relay interface.

4 Control systems

Operation 5
Operation overview 5.1
5 Operation
This chapter describes basic operation of WAVE Bioreactor 2/10 and WAVE Bioreactor
20/50 systems. For detailed instructions see relevant user documentation listed in
Section 8.2 Literature‚ on page 142.
5.1 Operation overview

WAVE system control
Control of operation, monitoring and key parameters during cell culturing is made by
embedded control systems in the Bioreactor base unit and/or external instrument
modules. For some functions there are different possibilities.
In Chapter 4 Control systems‚ on page 49 the basic functions of the control systems are
described.
Workflow
1 Preparations before start
2 Assemble system components
3 Calibrate the pH and DO probes
4 Fill Cellbag with culture medium
5 Inoculate the Cellbag

6 Set culturing conditions
7 Start culturing
8 Monitor and control culture conditions
9 Optional: sampling the culture
10 Optional: exchange culture medium
11 Optional: scaling up culture
12 Optional: perfusion culture
13 Stop culture and harvest
14 Procedures after a run

5 Operation
5.2 Preparations before starting a culture
5.2.1 Starting the system
Step Action
1 Make sure that any desired external instrument units are correctly
installed.
2 Turn the Bioreactor base unit power switch on (I). It is located at the rear
of the instrument.
3 Optional:
Turn desired installed external instrument units power switches on (I).
4 The Bioreactor base unit initializes.

The Top plate of the Bioreactor base unit starts tilting slowly towards the
rear. It contacts the rear safety switch (white touchbar) and then move
towards the front until it hits the front safety switch. The Top plate then
moves to the sampling position (front down) and stops.
Note: If the safety switch located on top of the base is hit, the rocking
motion stops and the rocking unit moves to a level position.
Note: If the safety switches have been hit, power must be turned off (O)
and on (I) to reset the safety switch.
5 After initialization, the main screen is displayed on the LCD screen (WAVE
Bioreactor 2/10) or Touch screen (WAVE Bioreactor 20/50).
5.2.2 Assembly of system
Step Action
1 WAVE Bioreactor 20/50, assemble Heater pad if not in place:

• Place the Heater pad(s) on the Bioreactor base unit Top plate with the
white side up.
• Plug in the Heater cable into the marked connector on the rear of the
instrument.

Operation 5
Preparations before starting a culture 5.2
Step Action
2 Assemble Cellbag holder and temperature sensor if not in place.

WAVE Bioreactor 2/10:
• Place the desired Cellbag holder on the Bioreactor base unit Top
plate. It should fit snugly with the heater cable to the back.
• Connect the temperature sensor in the Cellbag holder to the
connector on the back of the instrument.
WAVE Bioreactor 20/50:
• Place the desired Cellbag holder on the Heater pad.
• Slide the Cellbag holder towards the back and ensure the tabs are
engaged.
• Push the front down so that the catches engage.
• Connect the temperature sensor in the Cellbag holder to the HEATER
connector on the side of the instrument.
Note: To remove a Cellbag holder, push the front catches and slide
towards the front and up

5 Operation
Step Action
3 Assemble Cellbag(s)
For a description of Cellbags with inlets and valves, see Section 1.6.
• Select Cellbag size depending on experiment demands and
instrument setting. For Dual operation two Cellbags are used. See
Table 5-1.
• Remove the selected disposable Cellbag(s) from the protective plastic
over wrap bags.
For WAVE Bioreactor 20/50:
• Open the guides on the
Cellbag holder by lifting the
cam levers located on either
side of the Cellbag holder.
• Place the Cellbag into the
Cellbag holder and push each
of the two plastic rods into the
corresponding open guide.
Then press the cam levers
down to lock the Cellbag into
the Cellbag holder.
• Place the Cellbag on the
Cellbag holder with the
sampling ports towards the
front.
For WAVE Bioreactor 2/10:
• Lay Cellbag on the Cellbag holder and push the rod on each end into
the gray plastic clips located on either side.
The rods should snap into the clips and secure the Cellbag firmly in
the Cellbag holder.
Note: WAVE Bioreactor 20/50 is equipped with barcode reader. A warning message is
generated if no bag is detected or if the Cellbag barcode is not valid.
Using any Cellbag that does not have a valid barcode is not permitted on WAVE
equipment and will void any warranty or service claims.

Operation 5
Table 5-1. System configuration and available Cellbag sizes.
WAVE system Cellbag size
2L 10L 20L 22L 50L
WAVE Bioreactor 2/10 + +

- Cellbag holder KIT2EH
WAVE Bioreactor 20/50 + + +

- single operation and Cellbag holder KIT20EHT

- dual operation and Cellbag holder KIT20EHT

- single operation and Cellbag holder KIT50EHT
WAVE Bioreactor 20/50 +

- dual operation and Cellbag holder KIT50EHT
5.2.3 Connect aeration

For some system configurations and external instrument modules it is possible to set
CO2 and/or O2 concentration in combination with aeration.
Step Action
1 Connect the aeration air line from the Air out port located on the left side
of the instrument (dual mode systems has an additional Air out port
located on the right side of the instrument) to the inlet filter on the
Cellbag (Section 1.6).
2 Air in port (located under Air out port) options:

• Unconnected to aerate using room air.
• Connect to a gas source (CO2MIX20, O2MIX20 or other suitable gas
source < 0.2bar).
Gas is drawn in from the Air in port by an internal pump and pumped out
the Air out port to the Cellbag.
3 Make sure both inlet and outlet filter tubing clamps are open.

5 Operation
5.2.4 Connect Exhaust filter

heater(s)
The heater maintains the filter heater between 50°C to 60°C preventing condensation
in the filter and potential flow restriction.
Note: For Dual setup with two Cellbags on the same rocker, two exhaust filter heaters
are required.
Step Action
1 Insert the exhaust filter into the FLTHTR2 silicone heater.
2 Make sure the filter is snug.
3 Plug the filter heater cable into the heater connector(s) located on the
rear panel.
4 Check that the power ON light on each filter heater is lit.
5.2.5 Set operation control and

monitoring conditions
Operating conditions
recommendations
Each cell line and medium requires some optimization of operating conditions. The
rocking rate should be set at the minimum rate that provides mixing and oxygen
transfer without excessive foaming, see Table 5-2 for typical rocking and aerating rates.
Table 5-2. Typical operating conditions (WAVE Bioreactor 20/50 system).
Culture Cellbag Rocking rate (rpm) Aeration rate (lpm)

volume size
(liter) System 2/10 System 20/50 System 2/10 System 20/50
0.2 2L 10 to 15 10 0.1 to 0.2 0.1
1 2L 15 to 25 15 0.1 to 0.2 0.1
0.5 to 1 10L 10 to 15 10 0.1 to 0.2 0.1
5 10L 15 to 25 15 0.2 to 0.3 0.1 to 0.2
10 20L Not applicable 25 Not applicable 0.1 to 0.2
25 50L Not applicable 20 to 25 Not applicable 0.2 to 0.3
Operation 5
Rocking rate recommendations

• Increase the rocking rate if no wave is visible.
• Reduce the rocking rate if there is excessive foaming.
• Any rocking rate higher than 6 rocks per minute is sufficient for particle
suspension and bulk mixing.
• In general a rocking rate of 20 to 25 rocks per minute satisfies oxygen demands

6
for a cell density up to 5 × 10 cells/mL. However, cell population and metabolism
determines the necessary rocking rate for oxygen transfer.
• Measuring dissolved oxygen is the best way to determine the required rocking
rate. This may be done by off-line or on-line measurement. See Section 5.3.4.
Aeration rate recommendations

• Aeration rate (through the inlet air filter) has little effect on oxygen transfer.
Aeration rates should be kept to a minimum to reduce evaporation.
• Aeration rate may be adjusted to change the pCO2 or pH.
Recommendations to reduce foaming

• Foaming can be reduced by lowering the rocking angle.
• Typical rock angle is 6 to 8 degrees.
• For very foamy medium, the rock angle should be reduced to 4 to 5 degrees.
• For cells with high oxygen demand, such as insect cells, the rock angle can be
increased to 10 degrees.
• If extensive amount of foam is generated reduce the rocking speed. Note however
that is critical that the rocking speed be sufficient to generate a visible surface
wave.
• It is usual practice to add 0.01 mg/liter of pluronic F-68 to the culture medium to
minimize protein degradation associated with the effects of foaming. Most
commercial cell culture media already contains pluronic.
Excessive foaming also occurs if the Cellbag is not rigidly inflated. Check that you have
sufficient airflow and that the pressure relief valve is functioning
Set operating conditions

Set operating conditions for rocking and aeration and optionally for dissolved oxygen
measurement, control of pH, CO2 and O2 using the control systems for the system and
external instrument modules used. Main functions of control systems are described in
Chapter 4 Control systems‚ on page 49. For details, see relevant user documentation
listed in Section 8.2 Literature‚ on page 142.
5 Operation
5.3 Performing a run
5.3.1 Procedures before start

For instructions to monitor and set operating conditions for the current system
configuration, see relevant parts of Chapter 4 Control systems‚ on page 49.
Step Action
1 Calibrate pH and DO probes before each run, see Section 6.4 Calibration‚
on page 114.
2 Start rocking and aeration.
3 Verify that the Cellbag is firmly inflated and secured to the Cellbag holder.
4 Verify that the Cellbag holder is firmly latched on the Bioreactor base unit
Top plate. The Cellbag should be taut, but not creased.
5 Verify that air is released through the outlet pressure relief vent by
pressing down gently on the chamber and observing a release of air
through the outlet pressure relief valve.
5.3.2 Fill Cellbag with culture

medium
WARNING
for example 3 persons for 30 to 45 kg.
All lifting and moving must be performed in accordance with local
regulations.
Note: Continue airflow to the headspace to keep the Cellbag rigidly inflated. This is
important otherwise excessive foam is generated.
Step Action
1 Ensure that the Cellbag is inflated prior to filling with medium in order to
reduce foaming.
2 If rocking, stop the rocking.
Operation 5
Performing a run 5.3
Step Action
3 Connect, in an aseptic manner, tubing from the medium container to the

Cellbag using either the inlet tubing with a tube fusing device, or by
connecting a male luer fitting to one of the luer ports.
Note: Use of the luer fitting may require moving the Cellbag holder
containing the Cellbag into a biosafety cabinet.
4 Pump the desired volume of medium into the Cellbag.
5 Restart the rocking.
6 Adjust the rocking speed if required by changing the set point value.
• For rocking speed recommendations, see Operating conditions
recommendations‚ on page 100.
• Increase the speed if no wave is visible.
Reduce the speed if there is excessive foaming.
5.3.3 Inoculate the Cellbag

culture
Note: Continue airflow to the headspace to keep the Cellbag rigidly inflated.
Step Action
1 Stop the rocking motion.
2 Connect, in a sterile manner, tubing from the inoculum container to the

inlet tubing using either a tube fusing device or a male luer connector.
3 Pump the desired volume of inoculum into the Cellbag.

Note: Small inoculum volumes may be added by syringe through the
sampling port.
4 Restart the rocking motion.
5.3.4 Start culturing

Note: Ensure that the Cellbag is rigidly inflated at all times when rocking. An
underinflated bag may crack and leak. The Cellbag upper surface should be
drum-tight.
5 Operation
Note: Issues with the systems or set culturing conditions are acknowledged by an
alarm. An new alarm is indicated by a flashing ALARM button in the main screen
of the Touch screen. The Touch screen color also changes to red and an audible
beeper sounds. See Chapter 4 Control systems‚ on page 49 for handling Alarms.
Step Action
1 Make sure that desired operating conditions are set, see Operating
conditions recommendations‚ on page 100.
• It is possible to adjust rocking rate, rocking angle, aeration, and
temperature,
• For some system configurations it is also possible to monitor and/or
control dissolved oxygen, pH, CO2 concentration, O2 concentration
and conditions for performing perfusion culturing.
2 Start rocking, aeration and other desired monitor and/or control

functions.
5.3.5 Culture options
Sampling the Cellbag

Samples may be taken at least 50 times using the same sampling connector without
any danger of compromising sterility. The sample is also completely contained as no
aerosols are vented.
Step Action
1 Adjust the rocking rate if required.

At low rocking rates (<15 rpm) it is sometimes difficult to get a
representative samples due to settling. In this situation, the
recommended technique is to raise the rocking rate to 15 rpm 5 to 10
minutes prior to sampling. The rocking speed can be lowered after
sampling.
2 Remove the dust cap from the sampling

connector (see Section 1.6 for a Cellbag
description).
Operation 5
Step Action
3 Wipe the top of the sampling connector with

70% alcohol (or equivalent).
4 Using aseptic technique, attach a sterile

disposable syringe onto the connector.
5 Release the tubing clamp and withdraw a sample into the syringe. You
may need to push down on the Cellbag to force the liquid up the sample
tube.
6 Remove the syringe and wipe the top of the sampling connector again
with 70% ethanol and replace the dust cap.
7 Pinch the sampling connector tubing a few times to ensure that any
liquid in the tubing drains back into the Cellbag.
8 Close the tubing clamp.
Note: A standard syringe or luer connector may be used without a needle.
Note: The sampling device has a fairly small orifice. If you are working with large
microcarriers or large cell aggregates do not use the sampling connector.
5 Operation
Dissolved oxygen measurement

and control
WARNING
regulations.
The WAVE system is designed to provide an excess of oxygen for most culture systems.
To ensure desired levels of dissolved oxygen it is important to measure the levels of
dissolved oxygen and adjust the dissolved oxygen concentration if required.
Step Action
1 To measure the dissolved oxygen level:

• Off-line dissolved oxygen measurement can be made by taking
samples by syringe (see Sampling the Cellbag‚ on page 104) and
quickly determining the pO2 concentration using a blood-gas
analyzer.
• On-line dissolved oxygen measurement can be done using the micro-
thin flexible optical DO probe. The DO probe fits into the Oxywell2
sheath built into each Cellbag and can be inserted and removed
repeatedly for calibration without compromising sterility.
2 To adjust dissolved oxygen concentration:

• Using an integrated instrument module or an external instrument
modules which can provide any desired O2 concentration.
• To increase dissolved oxygen concentration, increase the rocking
speed.
Note: Dissolved CO2 concentrations are influenced in the inverse manner as dissolved
oxygen.
pH control
pH control can be achieved by:
• use of pH control module (WAVEPOD or standalone unit), which enables on-line
control of pH in the Cellbag.
• indirect control by control of the CO2 concentration (CO2 Control module,

integrated, WAVEPOD or standalone unit).
• use of appropriate buffers.
Operation 5
Scaling up culturing
WARNING
regulations.
Cellbags have a large range in operating volume. This allows an inoculum scaleup and
eliminates tedious sequential transfers. Start at low volume and add fresh medium to
the Cellbag as the cells grow. Up to a 1:10 expansion is possible in a single chamber.
A typical inoculum sequence is:
Step Action
1 Start with 100 mL medium in 2L Cellbag. Add inoculum.

6
2 When cells reach 2 × 10 cells/mL, add 300 mL medium to the Cellbag.
6
3 When cells again reach 2 × 10 cells/mL, add more medium to bring the
volume to 1 liter.
6
4 When these cells reach 2 × 10 cells/mL, transfer using a tube-fuser to a
20L Cellbag containing 2 liters of medium.
6
5 When the cells in the 20L Cellbag again reach 2 × 10 cells/mL, bring the
volume to 10 liters and continue cultivation at full volume.
In this sequence, only one transfer was made in going from 100 mL to 10 liters of
culture. This saves time and reduces the risk of contamination. This scheme can also be
used to scale-up inoculum for larger conventional bioreactors.
Culture medium exchange

WARNING
regulations.
5 Operation
Note: To avoid possible oxygen depletion, the culture medium exchange operation
should be performed in less than one hour.
Step Action
1 Stop the rocking and aeration and clamp off the inlet and outlet filters.
2 Remove the Cellbag holder from the rocking unit and place it in the
vertical position resting against a support. Allow the cells or microcarriers
10 to 15 minutes to settle.
3 Connect tubing to the harvest line on the Cellbag. The other end of this
tubing should be connected to a sterile collection vessel.
4 Using a peristaltic pump, remove the desired amount of supernatant

culture liquid by manually manipulating the flexible Cellbag wall.
5 Disconnect the tubing and reconnect to fresh medium to refill the

Cellbag.
6 Place the Cellbag holder back on the rocking unit.
7 Open the inlet and outlet filter clamps and restart aeration and rocking.
Perfusion culture
During perfusion culture cell-free harvest can be withdrawn and fresh medium can
be added continuously.
Requirements:
• Special Cellbags with internal cell retention filters
• WAVE Bioreactor 2/10 system with PERFCONT2E module or WAVE Bioreactor

20/50 system with BASE20/50EHT-L configuration.
It is recommended to contact a GE Healthcare applications specialist for advice

setting up perfusion culture.
For instructions to set perfusion culture operating conditions for the current
system configuration, see relevant parts of Chapter 4 Control systems‚ on page 49,
for control system instructions.
Operation 5
Procedures after a run 5.4
5.3.6 Stop culture and harvest

WARNING
regulations.
Note: The system has minimal turnaround time. Simply remove the completed batch
and place a new Cellbag holder and Cellbag on the instrument.
Step Action
1 Stop the rocking, aeration and other control and monitoring systems.
2 Clamp off the inlet and outlet filters of the Cellbag.
3 Disconnect the air inlet tube of the Cellbag.
4 Disconnect any other tubes and sensors still connected to the Cellbag.
5 Disconnect any connected cables to the Cellbag holder.
6 Remove the Cellbag holder containing the Cellbag from the base unit for
processing at any suitable location.
Tip: The Cellbag is a convenient harvest container. It is recommended to keep the

Cellbag on the Cellbag holder for easy storing and secure transport.
5.4 Procedures after a run

After completing a run and the removal of Cellbag and Cellbag holder.
Step Action
1 Turn the power switches of all used units off (O). Power switches are
generally located at the rear of respective unit.
2 If required clean system components (Section 6.1).
5 Operation
5.4 Procedures after a run
Maintenance 6
General 6.1
6 Maintenance
This chapter provides instructions for routine maintenance.
6.1 General
Regular maintenance of the WAVE systems is essential for reliable results.
WARNING
Electrical shock hazard. All repairs should be done by service
personnel authorized by GE Healthcare. Do not open any covers or
replace parts unless specifically stated in the user documentation.
WARNING
before performing any maintenance task.
NOTICE
Cleaning. Keep the instruments dry and clean. The instruments must be
turned off and unplugged before cleaning begins. Clean the exterior of
the instruments with a damp cloth with water and, if required, alcohol.
Do not use abrasive cleaners. Water should not be applied directly to the
instruments. Make sure that the instruments are completely dry before
plugging them in.
6 Maintenance
6.2 Safety switch inspection procedure
6.2 Safety switch inspection procedure

Required materials
3/8" ID × 5/8" OD, C-Flex or similar type tubing, 300 mm (12") long
Instruction
Step Action
1 Turn the power to the instrument on.
2 Set the instrument rocking speed to 10 rpm. Set the angle to 12 degrees
and turn rocker function on.
3 While the instrument is rocking, place one end of the 300 mm long piece
of tubing between the safety switch and the top rocker plate at front of
the instrument.
Result: The instrument should lightly squeeze the tubing, return to level
and then stop rocking. Within 30 seconds the background lighting of the
Touch screen should turn red and flash. The following message should be
displayed: “SAFETY BUMPER SWITCH HIT” “CYCLE POWER TO RESET”.
Note: If the instrument reacts differently than described, stop the
inspection and contact Technical support. Do not use the
equipment as the safety switches may not be working properly.
4 Repeat the procedure testing the rear safety switch.
Maintenance 6
Replacement of fuses 6.3
6.3 Replacement of fuses

General
WARNING
before replacing fuses.
WARNING
If a fuse requires repeated replacement, do not continue to use the
instrument. Contact an authorized service engineer.
Refer to Section 8.1 Specifications‚ on page 139 for information about the fuse types
and rating.
WARNING
For continued protection from fire hazard, replace only with same type
and rating of fuse.
Instruction
Follow the instruction below to replace fuses in WAVE Bioreactor 2/10, WAVE Bioreactor
20/50 and WAVEPOD.
Step Action
1 Disconnect the power cord from the instrument.
2 Pry off the fuse cover from the power entry module.
3 Remove and replace the fuses.
4 Place the fuse cover back in position.
5 Connect the power cord and turn on the instrument.
6 Maintenance
6.4 Calibration
6.4 Calibration
Before starting culturing, the pH probe and the DOOPT probe should be calibrated.
For further information on calibration of instrument modules, refer to WAVE Bioreactor
System 20/50EHT Operator Manual, WAVEPOD Operator Manual and WAVE Bioreactor
Instrumentation Manual.
pH calibration
Follow the instruction below to perform a single point calibration of the pH probe using
either WAVEPOD or pH20.
Step Actions
1 Power up the instrument.
2 Connect the pH probe to the PROBE connector on the WAVEPOD pH

module or to the SENSOR connector on pH20.
3 Fill the Cellbag with media and allow to equilibrate.
4 Measure the pH by sampling the Cellbag and using an external

calibrated pH meter.
5 Adjust the SPAN potentiometer until the reading on the controller

matches the grab sample reading. The SLOPE is adjusted at the factory
and should not need to be adjusted.
Result: The pH probe is now ready for use.
DO calibration
Calibration of the DOOPT probe requires a two-point method. Perform zero oxygen
calibration and 100% air saturation calibration according to the instructions below.
Preparations
Follow the instruction below to prepare for DO calibration.
Step Actions
1 Power up the instrument.
2 Connect the DOOPT probe to the PROBE connector on the WAVEPOD pH

module or to the PROBE connector on DOOPT20.
3 Open a new packet of zero oxygen calibration solution (ZERO OXYSOLN).

Immerse the probe tip in the solution and wait for the reading to stabilize.
4 Perform zero oxygen calibration and 100% air saturation calibration

according to the instructions below.
Maintenance 6
Calibration 6.4
Calibration using WAVEPOD

Follow the instruction below to perform a zero oxygen calibration using WAVEPOD.
Step Actions
1 On the DO OPTICAL control screen, press T/COMP. Enter the ambient

temperature using the pop-up keypad. Press ENT to set the new value.
2 Press the CALIB button. In the pop-up window, press the Set 0% button.
Result: A pop-up window displays SETTING 0%. When the pop-up
window closes the 0% oxygen point is set.
If the screen displays CAL FAIL, re-calibration is needed.
Follow the instruction below to perform a 100% air saturation calibration using
WAVEPOD.
Step Actions
1 Remove the probe from the zero oxygen solution. Rinse the probe in
water and shake dry. Allow the probe to equilibrate in air. Do not touch
the probe tip or attempt to dry it with a cloth.
2 When the reading is stable (within ±2%) in 1 to 2 minutes, press the CALIB
button. In the pop-up window, press the Set 0% button.
Result: A pop-up window displays SETTING 100%. When the pop-up
window closes the 100% oxygen point is set.
If the screen displays CAL FAIL, re-calibration is needed.
Calibration using DOOPT20

Follow the instruction below to perform a zero oxygen calibration using DOOPT20.
Step Actions
1 Press the PUSH FOR MENU button.

Result: The display shows --MENU-- 1.CALIB.

Result: The display shows SET ZERO -> NO.
3 Turn the PUSH FOR MENU button to change NO to YES.

Result: The display shows PUSH2SET P=54.95ª. (54.95ª is an example of
the phase angle. The numerical value shown can be slightly different).
6 Maintenance
6.4 Calibration
Step Actions
5 When the phase angle reading is stable (within ±0.25ª), press the PUSH
FOR MENU button.
Result: The display shows TRY0 0 (no of scans), 000.00% (actual reading).
If the display shows NEED 0% CAL, re-calibration is needed.
Follow the instruction below to perform a 100% air saturation calibration using
DOOPT20.
Step Actions
1 Remove the probe from the zero oxygen solution. Rinse the probe in
water and shake dry. Allow the probe to equilibrate in air. Do not touch
the probe tip or attempt to dry it with a cloth.
2 When the reading is stable (within ±2%) in 1 to 2 minutes, press the PUSH
FOR MENU button.
Result: The display shows --MENU-- 1.CALIB.
3 Press the PUSH FOR MENU button. twice

Result: The display shows SET 100% -> NO.
4 Turn the PUSH FOR MENU button to change NO to YES.

Result: The display shows PUSH2SET P=25.94ª. (25.94ª is an example of
the phase angle. The numerical value shown can be slightly different).
6 When the phase angle reading is stable (within ±0.25ª), press the PUSH
FOR MENU button.
Result: The display shows TRY0 0 (no of scans), 100.00% (actual reading).
If the display shows NEED 100% CAL, re-calibration is needed.
Troubleshooting 7
WAVE Bioreactor 2/10 7.1
7 Troubleshooting
7.1 WAVE Bioreactor 2/10

General problems
Error symptom Possible cause Corrective action
The Cellbag appears to Too high airflow Check that the airflow to the
be overinflated. bioreactor does not exceed
0.5 Lpm.
The Cellbag should be
inflated so that it Faulty relief valve Check that air is passing out of
requires force to the pressure relief valve: attach a
squeeze it. However, it short length of tubing to the
should not be so exhaust vent and immerse the
pressurized that tubing in water to a 1 cm depth.
creases form near the Bubbles should appear
attachment points. indicating flow. If no flow is
observed, remove the pressure
relief valve. The outlet filter may
be plugged and removing the
pressure relief valve may allow
continued operation.
Faulty Cellbag If the Cellbag continues to

overinflate, transfer the contents
to another Cellbag.
7 Troubleshooting
The Cellbag appears to Too low airflow Check that there is sufficient air
be underinflated. flow to the inlet filter.
An underinflated Inlet air supply Check that you have connected
Cellbag will generate wrongly connected the inlet air supply line to the
excessive foam and inlet filter (does not have the
poor mixing. pressure relief valve).
Missing pressure Check that the pressure relief

relief valve valve is present on the exhaust
filter.
Obstructed flow Check that air inlet and out flow

paths paths are unobstructed.
Clogged exhaust If the exhaust filter is clogged

filter then transfer the contents to
another Cellbag.
Excessive foaming in Poorly inflated Check that the Cellbag is rigidly

the Cellbag. Cellbag inflated. A poorly inflated bag
will foam rapidly.
Some amount of foam
is typical. Foam should Too high rocking rate If foam is still excessive after a
not cover more than few hours of operation, reduce
50% of the surface the rocking rate. Make sure that
area. the new rocking rate provides a
sufficient dissolved oxygen
concentration.
Too big rock angle Reduce the rock angle. The

bioreactor base units are factory
set to ±6° from the horizontal.
This angle has been found
optimal for most cell lines.
Troubleshooting 7
WAVE Bioreactor base unit
The rocking unit does Faulty safety Do not operate the instrument in
not initialize properly switches this condition. Contact Technical
support.
The error message The heater is Check that the heater cord is
HTROUT is displayed disconnected from firmly connected.
the WAVE Bioreactor
base unit
The temperature WARNING! Do not touch the

exceeds 60°C Cellbag holder.
Check that the sensor is
correctly installed and that a
Cellbag is present. In the event of
an overheat condition, the
system will reset automatically
once the heater cools down
sufficiently.
The error message The temperature Check that the temperature

RTDFAIL is displayed sensor is faulty or sensor is properly connected to
disconnected. This the rear panel connector.
will cause the heater If the problem persists, replace
to shut off. the temperature sensor.
Heating is not working Faulty rocking unit Check that the rocking unit is
and no error message rocking. The heater is
is displayed automatically turned off when
the rocking unit is not rocking.
Starting the rocking will
automatically switch the heater
on if temperature control is
enabled.
Poor temperature The temperature Make sure that the temperature

control. The sensor is not sensor is correctly installed.
temperature PV properly installed
(measured value) and
the SP (setpoint) is not The Cellbag does not Make sure that the Cellbag is
within 0.5°C. cover the correctly placed so that the
temperature sensor contents of the Cellbag cover the
temperature sensor.
7 Troubleshooting
Alarm messages
Message Description
HIPRES High bag pressure alarm
HTROUT Heater disconnected or over temperature alarm
RTDFAIL Temperature probe failed
SPDDEV Speed deviation from setpoint
AIRDEV Airflow deviation from setpoint
TMPDEV Temperature deviation from setpoint
WTERR Weighing system failure
LOWWT Low weight alarm
HIGHWT High weight alarm
NOFEED If the shot size is not fed after 20 minutes the alarm is triggered
and the perfusion is turned off.
NOHARV If the shot size is not harvested after 20 minutes the alarm is
triggered and the perfusion is turned off.
Troubleshooting 7

General problems
The Cellbag appears to Too high airflow Check that the airflow to the
be overinflated. bioreactor does not exceed 0.5
Lpm.
The Cellbag should be
inflated so that it Faulty relief valve Check that air is passing out of
requires force to the pressure relief valve: attach
squeeze it. However, it a short length of tubing to the
should not be so exhaust vent and immerse the
pressurized that tubing in water to a 1 cm depth.
creases form near the Bubbles should appear
attachment points. indicating flow. If no flow is
observed, remove the pressure
relief valve. The outlet filter may
be plugged and removing the
pressure relief valve may allow
continued operation.
Faulty Cellbag If the Cellbag continues to

overinflate, transfer the contents
to another Cellbag.
The Cellbag appears to Too low airflow Check that there is sufficient air
be underinflated. flow to the inlet filter.
An underinflated Inlet air supply Check that you have connected
Cellbag will generate wrongly connected the inlet air supply line to the
excessive foam and inlet filter (does not have the
poor mixing. pressure relief valve).
Missing pressure Check that the pressure relief

relief valve valve is present on the exhaust
vent filter.
Obstructed flow Check that air inlet and out flow

paths paths are unobstructed.
Clogged exhaust Transfer the contents to another

vent filter Cellbag.
7 Troubleshooting
Excessive foaming in Poorly inflated Check that the Cellbag is rigidly

the Cellbag. Cellbag inflated. A poorly inflated
Cellbag will foam rapidly.
Some amount of foam
is typical. Foam should Too high rocking rate If foam is still excessive after a
not cover more than few hours of operation, reduce
50% of the surface the rocking rate. Make sure that
area. the new rocking rate provides a
sufficient dissolved oxygen
concentration.
Too big rock angle Reduce the rock angle. The

WAVE Bioreactor base units are
factory set to ±6° from the
horizontal. This angle has been
found optimal for most cell lines.
Temperature control
The temperature The temperature Check that the temperature

screen displays a FAIL sensor is not sensor is properly connected.
message connected or faulty
The temperature is Adjust the temperature.

outside the range
0°C to 45°C
No heating The rocking unit is Check that the rocking unit is

not rocking rocking. The heater is
automatically turned off when
the rocking unit is not rocking.
This can be overridden from the
SETUP:HEATER menu.
Troubleshooting 7
Slow heating The heater cable is Check that the heater cord is
not connected firmly connected.
Remember, the heater
is intended to control The heater is not • Turn the heater on.
temperature using turned on
gentle heating. It takes • Check that the HTR ON
over 1 hour to heat 10 indicator is flashing.
liters from room • Check that the heater power
temperature to 37°C. bars are increasing or at
maximum.
• When using 20 L or larger
Cellbags, make sure that
both heaters are running
(LEFT+RGHT setting) on
HEATER control screen.
Wrong setpoint set Check that the setpoint is set

correctly.
The temperature The Cellbag does not Make sure that the temperature
control is not cover the sensor is properly positioned
functioning or the temperature sensor under the Cellbag.
displayed temperature
The temperature Check that the heater is correctly
appears to be
sensor is not positioned and that the Cellbag
incorrect.
properly installed holder is latched securely.
The temperature Calibrate the temperature

sensor needs sensor.
calibration
The HTR FAIL alarm is The temperature WARNING! Do not touch the
activated exceeds 60ªC Cellbag holder.
Check that the sensor is
correctly installed and that a
Cellbag is present. In the event of
an overheat condition, the
system will reset automatically
once the heater cools down
sufficiently.
The heater is Connect the heater to the WAVE

disconnected from Bioreactor base unit.
the bioreactor base
unit
7 Troubleshooting
The temperature Fluctuations in Use the lid to minimize the

fluctuates ambient air impact of ambient air
temperature movement.
Rocking unit
Rocking unit stops The rocking unit is Clear the obstruction and turn
rocking and the mechanically the power off and on to resume
SAFETY SWITCH restricted from the rocking.
TRIPPED alarm is moving
activated
Aeration controller
No airflow, and a high The airpump has Disconnect the Air in and Air out
pressure alarm is shut down due to tubing and locate the blockage.
activated high pressure
The airpump is running Blocked air tubing Disconnect the inlet and outlet
but the airflow shows connections on the side panels. If
zero the airflow is restored then check
if the Air in or Air out tubing is
blocked.
The airflow fluctuates, Restriction in the air Disconnect the inlet and outlet
and a high pressure tubing connections on the side panels. If
alarm is activated. the airflow is restored then check
for restrictions in the Air in or Air
Airflow should stay
out tubing.
constant within ± 0.02
Lpm. Clogged Cellbag Transfer the contents to another
exhaust vent filter Cellbag.
Troubleshooting 7
Weight controller
No weight reading LCELL20/50EHT not Check that the LCELL20/50EHT

installed option is installed and that the
sensor cable to the rear of the
bioreactor base unit is
connected correctly.
Weight option is not Verify that the WEIGHT option is

enabled enabled (SETUP:OPTIONS
screen). Go the TARE screen and
check that the gross weight is
positive.
Erratic weight display Mechanical Verify that the Cellbag holder is

obstructions to the firmly locked in position. Make
rocking unit sure that no tubing is pulling on
the Cellbag during rocking.
Check that the movement of the
rocking unit is not obstructed.
CO2 controller
No CO2 reading No CO2 module Verify that the bioreactor base

installed unit is type 20/50EHT-CO2.
Erratic CO2 control CO2 gas not properly Check that the CO2 gas to the
connected unit is connected and at the
correct pressure.
O2 controller
No O2 reading No O2 module Verify that the bioreactor base

installed unit is type 20/50EHT-O2.
Erratic O2 control O2 gas not properly Check that the O2 gas to the unit
connected is connected and at the correct
pressure.
7 Troubleshooting
Bar code reader
A message is displayed Faulty bar code Make sure that the Cellbag is
stating that no Cellbag reader valid and correctly installed.
is detected, that the If the problem persists, contact
Cellbag is incorrectly GE Healthcare Technical
installed in the holder, support.
or that the Cellbag bar
code is not valid.
Alarm messages
The table below lists the alarm messages that can appear on the
WAVE Bioreactor 20/50 Touch screen.
Alarm # Message Description
E1 INIT FAIL Machine failed to initialize on power up
E2 SPEED DEV Actual rocking speed does not match setpoint
E3 AIR DEV Actual airflow (left) does not match setpoint
E4 TEMP DEV Actual temperature (left) does not match setpoint
E5 WEIGHT Actual weight does not match setpoint

DEV
E6 TEMPFAIL Temperature sensor (left) failed or unplugged
E7 HTR FAIL Heater unplugged or over temperature
E8 ROCKFAIL Rocker not rocking
E9 COMMFAIL Internal communications failure. Cycle power to retry.
E10 MTRFAIL Rocking motor failure
E11 HIPRALM_L High bag (left) pressure.
E12 EMERSTOP Safety bumper activated and stopped rocker
E13 CO2FAIL CO2 sensor fail (2050EHT-CO2 only)
E14 PANTFAIL Not used for bioreactor
E15 RTEMPFAIL Right side temperature sensor failed or unplugged

(2050EHTD)
Troubleshooting 7
E16 RTEMPDEV Actual temperature (right) does not match setpoint

(2050EHTD)
E17 RAIRDEV Actual airflow (right) does not match setpoint

(2050EHTD)
E18 CO2DEV CO2 deviation (2050EHT-CO2 only)
E19 HIPRALM_R High bag (right) pressure (2050EHTD only)
E20 LOADFAIL Weight sensor failed (only when LCELL20/50EHT is

installed)
E21 LOWWTALM Low weight alarm (only when LCELL20/50EHT is

installed)
E22 HIWTALM High weight alarm (only when LCELL20/50EHT is

installed)
E23 HARVALM Harvest alarm (only when LCELL20/50EHT is installed)
E24 FEEDALM Feed alarm (only when LCELL20/50EHT is installed)
E25 LOCO2PR Low CO2 supply pressure (2050EHT-CO2 only)
E26 CODE_FAIL Bar code reader failed
E27 O2FAIL O2 sensor fail (2050EHT-O2 only)
E28 LOO2PR Low O2 supply pressure (2050EHT-O2 only)
E29 O2DEV O2 deviation (2050EHT-O2 only)
E30 LOAD_OVRG Weight exceeds sensor range
7 Troubleshooting
7.3 WAVEPOD
7.3 WAVEPOD
General problems
The table below gives a brief guide of how to solve general problems that might occur
when using WAVEPOD.
Display stays blank No power Check the Ethernet status LED. If

connected it is not lit, check the fuses and
the power cord.
The message: BASE This message is Contact GE Healthcare Technical

NOT FOUND is displayed support.
displayed repeatedly momentarily on
startup and if
communications to
the connected base
are lost. If the
message stays on
then internal
communications to
WAVEPOD control
modules have been
lost.
Dissolved oxygen controller

The table below lists the error codes for the optical dissolved oxygen probe.
Error code Description
0 ADC overflow
1 High ambient light
4 Low amplitude
Troubleshooting 7
WAVEPOD 7.3
The table below gives a brief guide of how to solve problems that might occur when
using the DO (Dissolved Oxygen) controller.
Low amplitude alarm Probe not properly Check that the probe is
connected or faulty connected correctly and make
probe sure that the plug is secure. Cycle
power to the instrument to reset.
On the WAVEPOD Touch screen
overview screen, press DO OPT
to display the DETAIL screen for
the display for the DO controller.
Check the signal amplitude. Less
than 2000 indicate a bad probe.
If the error persists, replace the
probe.
Fluctuating reading Some fluctuation -

(±5%), especially at
high DO levels is
normal
Probe adjustment is Faulty probe The probe should be readjusted

drifting every 2 to 4 days to compensate
for drift. If the drift is greater than
10%, the probe should be
replaced.
CO2 MIX
using the CO2 controller.
The display shows a A minor deviation If the reading is not within the
CO2 concentration in from the expected tolerance, the CO2 offset can be
air which deviates from value is not unusual. adjusted, see WAVEPOD
the expected value The tolerance of the Operator Manual.
(0.0%). CO2 sensor is
±0.38% in the range
0% to 7.5% CO2.
7 Troubleshooting
7.3 WAVEPOD
CO2 reading keeps The CO2 gas supply Check that the green CO2 inlet
drifting down pressure is too low pressure light is lit. The CO2
supply pressure must between
10 and 15 psig.
Faulty CO2 control The CO2 supply is not Verify that CO2 is connected to
properly connected the correct inlet port. Check that
the air pump is on and that the
flow rate is between 0.1 and 0.5
liters/minute. Check that the CO2
switch is on.
Wrong setpoint set Check the setpoint on the

controller.
No airflow reading Obstructed airflow Check that airflow to the Cellbag

is not obstructed.
No air flow Make sure that the pump is

working by disconnecting the
MIX OUT coupling on the
instrument.
The Cellbag does not No airflow Verify that there is airflow to the
stay inflated Cellbag.
Faulty check valve Check that the check valve on

the Cellbag is correctly installed.
Close off the Cellbag outlet using
the pinch clamp and see if the
Cellbag inflates. Then confirm
that the check valve is working
by connecting a short length of
tubing to the check valve outlet
and dip it into water. Tubing
should be immersed to a depth
of 5 to 10 mm. Opening the outlet
pinch valve should cause
bubbles to appear and bubbles
should continue to flow as the
flow and pressure stabilizes.
Troubleshooting 7
WAVEPOD 7.3
O2 MIX
using the O2 controller.
The display shows a O2 A minor deviation If the reading is not within the
concentration in air from the expected tolerance, the O2 offset can be
which deviates from value is not unusual. adjusted, see WAVEPOD
the expected value The tolerance of the Operator Manual.
(21.0%). O2 sensor is ±0.6% in
the range 21% to
40% O2.
O2 reading keeps The O2 gas supply Check that the green O2 inlet
drifting down pressure is too low pressure light is lit. The O2 supply
pressure must between 10 and
15 psig.
Faulty O2 control The O2 supply is not Verify that O2 is connected to the

properly connected correct inlet port. Check that the
O2 supply pressure is set to 0.7
bar minimum. Check that the air
pump is on and that the flow rate
is between 0.1 and 0.5 liters/
minute. Check that the O2 switch
is on.

controller.
No airflow reading Obstructed airflow Check that airflow to the Cellbag

is not obstructed.
No air flow Make sure that pump is working

by disconnecting the MIX OUT
coupling on the instrument.
Alarm messages
The table below lists the alarm messages that can appear on the WAVEPOD Touch
screen.
01 BAY1&3 FAIL Communications with bay 1 and 3 lost
02 BAY2&4 FAIL Communications with bay 2 and 4 lost
7 Troubleshooting
7.3 WAVEPOD
03 BASE COM FAIL Communications with bioreactor base unit lost
10 PH_DEV pH deviates from setpoint
11 PH_FAIL pH sensor failed or disconnected
12 PHCOM_FAIL pH amplifier not responding
20 DO_DEV DO deviates from setpoint
21 DO_FAIL DO sensor failed or disconnected
30 INIT_FAIL PLC initialization failure
31 AIR_DEV Airflow deviates from setpoint
32 OUTHIPRALM MIX OUT pressure too high
40 O2_DEV O2 deviates from setpoint
41 CO2_DEV CO2 deviates from setpoint
42 O2_FAIL O2 sensor failed
43 CO2_FAIL CO2 sensor failed
44 LOWO2PR Oxygen supply pressure low
45 LOWCO2PR CO2 supply pressure low
46 O2MINLMT O2 remote setpoint at minimum limit
47 O2MAXLMT O2 remote setpoint at maximum limit
48 CO2MAXLMT CO2 remote setpoint at maximum limit
49 CO2MINLMT CO2 remote setpoint at minimum limit
50 RPMMINLMT Speed remote setpoint at minimum limit
51 RPMMAXLMT Speed remote setpoint at maximum limit
Troubleshooting 7

DOOPT20
Not getting 0% Probes not properly Check that the DOOPT probe is
connected disconnected and that the
temperature probe is connected
(red LED is not lit).
Not getting 100% Faulty DOOPT probe Check that the temperature
probe is connected (red LED is
not lit). Check that zero is
obtained when the DOOPT probe
is disconnected. If it is still not
possible to get zero with the
SPAN knob, then replace the
DOOPT probe.
Fluctuation reading Some fluctuation The fluctuation can be damped

(±5%), especially at by adjusting the filter time.
high DO levels is
normal
Probe calibration is The probe is not Re-calibrate the DOOPT probe

drifting properly calibrated daily to compensate for drift. If
the drift is greater that 10% per
day, the probe should be
replaced.
Low amplitude alarm The probe is not Check that the probe is
properly connected connected correctly and make
sure that the plug is secure. Cycle
power to the instrument to reset.
The probe amplitude On the WAVEPOD Touch screen

is low overview screen, press DO OPT
to display the DETAIL screen for
the DO controller. Check the
signal amplitude. Less than 2000
indicate a bad probe.
If the error persists and the
amplitude is below 2000, replace
the probe.
7 Troubleshooting
No response from The internal PC To check operation, turn off

display board is defective power to DOOPT20 and
disconnect the probe. Turn on
power to DOOPT20, look directly
into the probe connector. A
pulsing light about once a
second should be flashing. If no
pulsing light appears, the unit
internal PC board requires
replacement. Contact GE
Healthcare Technical support.
NEED 0% CAL or NEED The probe requires Calibrate the probe. If the
100% CAL is displayed calibration problem persists, replace the
probe.
Unable to select If you are unable to Contact GE Healthcare Technical

menus use the PUSH FOR support.
MENU button, the
switch is defective
The display shows a A minor deviation If the reading is not within the
CO2 concentration in from the expected tolerance, the CO2 offset can be
air which deviates from value is not unusual. adjusted, see WAVE Bioreactor
the expected value The tolerance of the Instrumentation Manual.
(0.0%). CO2 sensor is
±0.38% in the range
0% to 7.5% CO2.
CO2 reading keeps The CO2 gas supply Check that the green CO2 inlet
drifting down pressure is too low pressure light is lit. The CO2
supply pressure must between
10 and 15 psig.
Troubleshooting 7
Faulty CO2 control The CO2 supply is not Verify that CO2 is connected to
properly connected the correct inlet port. Check that
the air pump is on and that the
flow rate is between 0.1 and 0.5
liters/minute. Check that the CO2
switch is on.

controller.
No reading on Obstructed airflow Check that airflow to the Cellbag

rotameter is not obstructed. Verify that the
pump is working by
(Applies to -R version
disconnecting the MIX OUT
only)
The Cellbag does not No air flow Verify that there is airflow to the
stay inflated Cellbag.
Faulty check valve Check that the check valve on

the Cellbag is correctly installed.
Close off the Cellbag outlet using
the pinch clamp and see if the
Cellbag inflates. Then confirm
that the check valve is working
by connecting a short length of
tubing to the check valve outlet
and dip it into water. Tubing
should be immersed to a depth
of 5 to 10 mm. Opening the outlet
pinch valve should cause
bubbles to appear and bubbles
should continue to flow as the
flow and pressure stabilizes.
The display shows a O2 A minor deviation If the reading is not within the
concentration in air from the expected tolerance, the O2 offset can be
which deviates from value is not unusual. adjusted, see WAVE Bioreactor
the expected value The tolerance of the Instrumentation Manual.
(21.0%). O2 sensor is ±0.6% in
the range 21% to
40% O2.
7 Troubleshooting
O2 reading keeps The O2 gas supply Check that the green O2 inlet
drifting down pressure is too low pressure light is lit. The O2 supply
pressure must between 10 and
15 psig.
Faulty O2 control The CO2 supply is not Verify that O2 is connected to the
properly connected correct inlet port. Check that the
O2 supply pressure is set to 0.7
bar minimum. Check that the air
pump is on and that the flow rate
is between 0.1 and 0.5 liters/
minute. Check that the O2 switch
is on.

controller.
No reading on Obstructed airflow Check that airflow to the Cellbag

rotameter is not obstructed. Verify that
pump is working by
(Applies to -R version
disconnecting the MIX OUT
only)
PUMP20
The PUMP ON button The pump is not • Make sure that the PUMP20
on the front panel of powered is plugged in.
PUMP20 is not lit
• If using a WAVE Bioreactor
20/50 for power, check that
the bioreactor is turned on.
• For standalone use, make
sure that PUMP20 is properly
connected to an external
power supply.
PUMP20 does not run A shorting plug is not For standalone use, make sure
connected that a shorting plug is connected
to the DB9 connector on the rear
panel of PUMP20
Troubleshooting 7
The pump speed does The speed settings • Check the speed settings on
not change are not correct the front panel of the
PUMP20.
• Press the RUN button to run
the pump all full speed
regardless of the speed
setting.
pH20
The message Err is The pH probe is not Make sure the pH probe is
displayed on the connected properly connected.
screen
The pH probe is Replace the pH electrode if it is
faulty damaged.
The acid pump does The pump is not Make sure that the switches on
not run turned on the pump are on and that the
pump is powered.
The pump is not Make sure that the acid pump is

connected to pH20 connected to the ACID
connector on the front of pH20.
The acid pump is not Make sure that the ACID ENBL
enabled to run switch on pH20 is pressed and lit
automatically up.
The set checkpoint is Check that the setpoint is set

not correct correctly.
7 Troubleshooting
The base pump does The pump is not Make sure that the switches on
not run turned on the pump are on and that the
pump is powered.
The pump is not Make sure that the base pump is

connected to pH20 connected to the BASE
connector on the front of pH20.
The base pump is Make sure that the BASE ENBL

not enabled to run switch on pH20 is pressed and lit
automatically up.
The set checkpoint is Check that the setpoint is set

not correct correctly.
pH overshoots The gain value is too Adjust the gain to a lower value.
high
The cycle time is to If the next addition occurs before

short the pH stabilizes, increase the
cycle time.
Constant pH is not Obstructed tubing Check that the acid pump tubing
maintained is open.
The pump is turned Check that the pump is running.

off
The gain value is too If the pH does not change

low significantly upon each addition,
adjust the gain to higher value.
Reference information 8
Specifications 8.1
8 Reference information
This chapter contains technical data, regulatory and other information.
8.1 Specifications
Parameter Value
Ingression protection IP 20
Supply voltage 100-120 V AC or 220-240 V AC,

50 to 60 Hz
Power 145/210 VA
Maximum current 4A
Fuse specification T 4AL, 250V
Dimensions (h × w × d) 160 × 230 × 330 mm

with Cellbag holder:
200 (300 incl. Cellbag) × 489 × 330
Weight 4.2 kg
Acoustic noise level < 70 dB A
Ambient temperature 0ªC to 50ªC
Storage temperature -40ªC to +80ªC
Relative humidity tolerance < 95%, non-condensing
8.1 Specifications
Parameter Value
Ingression protection IP 20

50 to 60 Hz
Power 630 VA
Dimensions (h × w × d) • 179 × 573 × 46 mm
• With Cellbag holder KIT20EHT:

254 × 711 × 575
• With Cellbag holder KIT50EHT:

254 × 775 × 700
Weight 16 kg
Storage temperature -40ªC to +80ªC
Relative humidity tolerance < 95%, non-condensing
Specifications 8.1
WAVEPOD
Parameter Value
Ingression protection IP30

50 to 60 Hz
Power 200 VA
Weight 8.6 kg
Storage temperature -20°C to +60°C
Relative humidity tolerance 10% to 90%, non-condensing
DOOPT20
Parameter Value

50 to 60 Hz
Weight 1.5 kg
Parameter Value

50 to 60 Hz
Weight 2.7 kg
8.2 Literature
Parameter Value

50 to 60 Hz
Weight 2.7 kg
PUMP20
Parameter Value

50 to 60 Hz
Weight 1.0 kg
pH20
Parameter Value

50 to 60 Hz
Weight 1.5 kg
8.2 Literature
For further information regarding WAVE Bioreactor 2/10, WAVE Bioreactor 20/50,
WAVEPOD, and the external instrument modules, refer to the following:
• WAVE Bioreactor System BASE 2/10 EH Operator Manual
• WAVE Bioreactor System 20/50EHT Operator Manual
• WAVEPOD Operator Manual
• WAVE Bioreactor Instrumentation Manual
Ordering information 8.3
8.3 Ordering information

For ordering information, visit www.gelifesciences.com/wave.
8.3 Ordering information
For local office contact information, visit GE, imagination at work and GE monogram are trademarks of General
Electric Company.
www.gelifesciences.com/contact , Wave Bioreactor, WAVEPOD, and Cellbag are trademarks of GE
Healthcare companies.
GE Healthcare Bio-Sciences AB All third party trademarks are the property of their respective owners.
Björkgatan 30 © 2009 General Electric Company—All rights reserved.
751 84 Uppsala First published Nov. 2009
Sweden All goods and services are sold subject to the terms and conditions of sale
of the company within GE Healthcare which supplies them. A copy of these
terms and conditions is available on request. Contact your local GE
Healthcare representative for the most current information.
www.gelifesciences.com/wave
GE Healthcare UK Ltd
Amersham Place, Little Chalfont, Buckinghamshire, HP7 9NA, UK
GE Healthcare Bio-Sciences Corp

800 Centennial Avenue, P.O. Box 1327, Piscataway, NJ 08855-1327, USA
GE Healthcare Europe GmbH

Munzinger Strasse 5, D-79111 Freiburg, Germany
GE Healthcare Japan Corporation

Sanken Bldg. 3-25-1, Hyakunincho, Shinjuku-ku, Tokyo 169-0073, Japan
imagination at work
28-9641-23 AA 11/2009

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WAVE Bioreactor™ 2/10

2 Safety instructions ................................................................ 25

WAVE Bioreactor 2/10 and 20/50 systems Operating Instructions 28-9641-23 AA 3

8 Reference information ....................................................... 139

4 WAVE Bioreactor 2/10 and 20/50 systems Operating Instructions 28-9641-23 AA

WAVE Bioreactor 2/10 and 20/50 systems Operating Instructions 28-9641-23 AA 5

1.1 Important user information

6 WAVE Bioreactor 2/10 and 20/50 systems Operating Instructions 28-9641-23 AA

Notes and tips

1.2 Regulatory information

Name and address of manufacturer GE Healthcare Bio-Sciences AB,

Name and ID of notified body INTERTEK SEMKO AB, NB 0413

Place and date of declaration Uppsala, Sweden, Nov 2009

Identity of person authorized to sign DoC See EC Declaration of Conformity.

2006/42/EC Machinery Directive (MD)

2006/95/EC Low Voltage Directive (LVD)

2004/108/EC ElectroMagnetic Compatibility (EMC) Directive

WAVE Bioreactor 2/10 and 20/50 systems Operating Instructions 28-9641-23 AA 7

Standard Description Notes

EN 61010-1, Safety requirements for electrical

EN 61326-1 EMC emissions and immunity Harmonized with

EN-ISO Safety of machinery – Basic concepts, Harmonized with

EN-ISO Safety of machinery – Principles of Harmonized with

• connected to other CE-marked instruments, or

8 WAVE Bioreactor 2/10 and 20/50 systems Operating Instructions 28-9641-23 AA

1.3 WAVE system configurations

• WAVEPOD with pH control module and PUMP20, or

• the external instrument modules pH20 and PUMP20.

• Indirect control of pH by control of the CO2 concentration. This requires:

• Integrated CO2 control module (as in BASE20/50EHT-CO2), or

• WAVEPOD with CO2 control module, or

• the external instrument modules CO2MIX20 or CO2MIX20-R.

WAVE Bioreactor 2/10 and 20/50 systems Operating Instructions 28-9641-23 AA 9

Table 1-1. Examples of WAVE Bioreactor system configurations. Note that

System configuration Control parameters

WAVE Bioreactor 2/10 system

BASE2/10EH pH20 and PUMP20 + + +

WAVE Bioreactor 20/50 system

BASE20/50EHT POD-R/L PH CO2 + + + +

BASE20/50EHT POD-R/L DOOPT O2 + + + +

BASE20/50EHT POD-R/L PH DOOPT CO2 O2 + + + + + +

BASE20/50EHT-L POD-R/L PH DOOPT CO2 O2 + + + + + + +

BASE20/50EHT pH20 and PUMP20 + + +

10 WAVE Bioreactor 2/10 and 20/50 systems Operating Instructions 28-9641-23 AA

1.4 WAVE Bioreactor 2/10 system

• Pre-sterilized disposable Cellbag (ordered separately)

• Perfusion controller (optional)

• External instrument modules (optional)

1 Bioreactor base unit

2 Front panel with controls

6 Perfusion controller (optional)

WAVE Bioreactor 2/10 and 20/50 systems Operating Instructions 28-9641-23 AA 11

1.5 WAVE Bioreactor 20/50 system

• Pre-sterilized disposable Cellbag (ordered separately)

• External instrument modules (optional)

1 Bioreactor base unit

2 Protective bellows, rocking mechanism

4 Cellbag holder (one of several options shown)

5 Detachable Touch screen

12 WAVE Bioreactor 2/10 and 20/50 systems Operating Instructions 28-9641-23 AA

3 Outlet air filter

4 Inlet air filter