cytiva ReadyToProcess WAVE 25 User manual

ReadyToProcess WAVE 25
User Manual
cytiva.com

Table of Contents
1Introduction ........................................................................................................ 5
1.1 Important user information ....................................................................................................................... 6
1.2 About this manual ......................................................................................................................................... 8
1.3 Associated documentation ....................................................................................................................... 9
1.4 Abbreviations .................................................................................................................................................. 10
2 System description ............................................................................................ 11
2.1 System overview ............................................................................................................................................ 12
2.2 ReadyToProcess WAVE 25 rocker .......................................................................................................... 16
2.3 ReadyToProcess CBCU .............................................................................................................................. 24
2.4 ReadyToProcess Pump 25 ......................................................................................................................... 27
2.5 Cellbag bioreactor ........................................................................................................................................ 29
2.6 UNICORN software overview .................................................................................................................... 32
2.6.1 General UNICORN operation .................................................................................................................. 33
2.6.2 UNICORN help ............................................................................................................................................... 34
3 The UNICORN software ...................................................................................... 36
3.1 Administration ................................................................................................................................................ 37
3.2 System control ................................................................................................................................................ 38
3.3 Methods in UNICORN .................................................................................................................................. 46
3.3.1 Method editor ................................................................................................................................................. 47
3.3.2 Method creation ............................................................................................................................................ 49
3.3.3 Work with methods ..................................................................................................................................... 53
3.3.4 Text instructions ........................................................................................................................................... 59
3.3.5 Save a method ............................................................................................................................................... 67
3.3.6 Scouting ............................................................................................................................................................ 71
3.3.7 Method queues .............................................................................................................................................. 77
3.4 Evaluation in UNICORN .............................................................................................................................. 82
3.4.1 Evaluation ........................................................................................................................................................ 83
3.4.2 Open and view results ................................................................................................................................ 85
3.4.3 Run documentation ..................................................................................................................................... 89
3.4.4 Generate and print a predefined report format .............................................................................. 94
3.4.5 Create a new report format ...................................................................................................................... 96
3.4.6 Edit an existing report format ................................................................................................................. 105
4 System control description ............................................................................... 107
4.1 Single and dual operation modes ............................................................................................................ 108
4.2 Temperature measurement and control .............................................................................................. 109
4.3 pH and DO measurement and control .................................................................................................. 110
4.3.1 pH and DO measurement principles .................................................................................................... 111
4.3.2 pH and DO control principles ................................................................................................................... 113
4.4 pH control ......................................................................................................................................................... 115
4.4.1 pH control schemes ..................................................................................................................................... 116
4.4.2 CO2 control mode ........................................................................................................................................ 118
4.4.3 Acid/Base control mode ............................................................................................................................ 120
4.4.4 pH control transition delays ..................................................................................................................... 125
4.5 DO control ........................................................................................................................................................ 126
4.5.1 DO control schemes .................................................................................................................................... 127
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4.5.2 O2 control mode ........................................................................................................................................... 128
4.5.3 Speed control mode .................................................................................................................................... 130
4.5.4 O2/Speed control scheme ........................................................................................................................ 131
4.6 Media control .................................................................................................................................................. 132
4.6.1 Weight measurement ................................................................................................................................ 133
4.6.2 Media addition ............................................................................................................................................... 134
4.6.3 Perfusion .......................................................................................................................................................... 135
4.6.4 Deviation alarm ............................................................................................................................................. 139
4.6.5 Media control inactivation ........................................................................................................................ 140
4.7 Recommended operating conditions .................................................................................................... 141
4.8 System verification ....................................................................................................................................... 144
5 Operation ............................................................................................................ 149
5.1 Set up the system .......................................................................................................................................... 150
5.1.1 Select the tray and Cellbag bioreactor ............................................................................................... 151
5.1.2 Attach and detach tray .............................................................................................................................. 152
5.1.3 Prepare pH and DO sensors ..................................................................................................................... 155
5.1.4 Attach the Cellbag bioreactor ................................................................................................................ 158
5.1.5 Prepare the pump ........................................................................................................................................ 160
5.1.6 Connect gas to the system ....................................................................................................................... 165
5.1.7 Connect the filter heater to the rocker ................................................................................................ 169
5.2 Start and configure the system ................................................................................................................ 170
5.2.1 Start the system and log on to UNICORN ......................................................................................... 171
5.2.2 Connect to the system ............................................................................................................................... 172
5.2.3 Configure system properties ................................................................................................................... 174
5.2.4 Configure system settings ........................................................................................................................ 176
5.2.5 Start a run ........................................................................................................................................................ 182
5.3 Prepare for cultivation ................................................................................................................................. 187
5.3.1 Inflate the Cellbag bioreactor ................................................................................................................. 188
5.3.2 Adjust pump parameters .......................................................................................................................... 189
5.3.3 Final checks before cultivation ............................................................................................................... 190
5.3.4 Add and equilibrate culture medium ................................................................................................... 191
5.3.5 Prepare the sensors .................................................................................................................................... 194
5.4 Perform cultivation ....................................................................................................................................... 197
5.4.1 Inoculate the culture ................................................................................................................................... 198
5.4.2 Monitor and control the run ..................................................................................................................... 199
5.4.3 End a run ........................................................................................................................................................... 206
6 Maintenance ....................................................................................................... 208
6.1 Calibration ........................................................................................................................................................ 209
6.2 Pump calibration ............................................................................................................................................ 211
6.3 Cleaning ............................................................................................................................................................ 214
7 Troubleshooting ................................................................................................. 215
7.1 ReadyToProcess WAVE 25 system ........................................................................................................ 216
7.2 ReadyToProcess WAVE 25 rocker .......................................................................................................... 217
7.3 ReadyToProcess CBCU .............................................................................................................................. 221
7.4 ReadyToProcess Pump 25 ......................................................................................................................... 233
7.5 Cellbag bioreactor ........................................................................................................................................ 234
7.6 Software problems ........................................................................................................................................ 236
7.6.1 Troubleshooting Method editor .............................................................................................................. 237
7.6.2 UNICORN System Control ........................................................................................................................ 239
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ReadyToProcess WAVE 25 User Manual 29009598 AD 3

7.6.3 Troubleshooting Evaluation ..................................................................................................................... 242
8 Reference information ....................................................................................... 243
8.1 System specifications .................................................................................................................................. 244
8.2 Component specifications ......................................................................................................................... 245
8.3 Client computer specifications ................................................................................................................ 248
8.4 Chemical resistance ..................................................................................................................................... 250
8.5 Control settings .............................................................................................................................................. 251
8.5.1 Rocking control .............................................................................................................................................. 252
8.5.2 Heating control .............................................................................................................................................. 255
8.5.3 Gas flow control ............................................................................................................................................. 256
8.5.4 CO2 mix control ............................................................................................................................................. 258
8.5.5 O2 mix control ................................................................................................................................................ 259
8.5.6 Pump control .................................................................................................................................................. 260
8.5.7 pH measurement ......................................................................................................................................... 263
8.5.8 pH control ........................................................................................................................................................ 265
8.5.9 DO measurement ......................................................................................................................................... 272
8.5.10 DO control ........................................................................................................................................................ 274
8.5.11 Media control ................................................................................................................................................. 278
8.6 Digital and analog I/O connections ......................................................................................................... 280
8.7 Ordering information .................................................................................................................................. 283
Index ........................................................................................................................... 285
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4 ReadyToProcess WAVE 25 User Manual 29009598 AD

1 Introduction
About this chapter
This chapter includes important user information, intended use of the system, and lists
of important concepts and user documentation.
In this chapter
Section See page
1.1 Important user information 6
1.2 About this manual 8
1.3 Associated documentation 9
1.4 Abbreviations 10
1 Introduction
ReadyToProcess WAVE 25 User Manual 29009598 AD 5

1.1 Important user information
Read the Operating Instructions
before operating the product
All users must read the entire separate Operating Instructions before instal-
ling, operating, or maintaining the product.
Always keep the Operating Instructions at hand when operating the product.
Do not operate the product in any other way than described in the user documenta-
tion. 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 ReadyToProcess™ WAVE™ 25 system is intended to be used as laboratory and
manufacturing equipment for cell cultivation. The system shall not be used for clinical
or diagnostic purposes.
Prerequisites
In order to operate ReadyToProcess WAVE 25 in the way it is intended:
•you have a general understanding of how the client computer and Microsoft®
Windows® operating systems work.
•you are acquainted with the use of general laboratory equipment and with handling
of biological materials.
•you have read and understood the Safety instructions chapter in the Operating
Instructions.
•the system is installed according to the instructions in the Operating Instructions.
•a user account has been created according to UNICORN Administration and
Technical manual.
Safety notices
This user documentation contains safety notices (WARNING, CAUTION, and NOTICE)
concerning the safe use of the product. See definitions below.
1 Introduction
1.1 Important user information
6 ReadyToProcess WAVE 25 User Manual 29009598 AD

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.
1 Introduction
1.1 Important user information
ReadyToProcess WAVE 25 User Manual 29009598 AD 7

1.2 About this manual
Purpose of this manual
The User Manual provides you with instructions and information how to run the
ReadyToProcess WAVE 25 bioreactor system. It also includes relevant guidance for
practical handling and maintenance of the system units.
Scope of this document
This manual covers the ReadyToProcess WAVE 25 system, including the rocker, gas
mixer and DO/pH controller (CBCU), pump, UNICORN™ software and accessories.
Typographical conventions
Software items are identified in the text by bold italic text.
Hardware items are identified in the text by bold text.
In electronic format, references in italics are clickable hyperlinks.
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 proce-
dures.
1 Introduction
1.2 About this manual
8 ReadyToProcess WAVE 25 User Manual 29009598 AD

1.3 Associated documentation
Introduction
This section describes the user documentation that is delivered with the product, and
how to find related literature that can be downloaded or ordered from Cytiva.
User documentation for
ReadyToProcess WAVE 25
The table below describes the user documentation for ReadyToProcess WAVE 25,
which is available from the Help menu in UNICORN or on the user documentation CD.
Document Main contents
ReadyToProcess WAVE 25
Operating Instructions
(29009597)
Instructions needed to install, operate and main-
tain ReadyToProcess WAVE 25 in a safe way.
Includes basic UNICORN 7.x system control func-
tions.
ReadyToProcess WAVE 25
User Manual (29009598)
Detailed system descriptions and instructions on
how to run, maintain and troubleshoot
ReadyToProcess WAVE 25. Includes UNICORN 7.x
system control functions, method creation and
handling, together with evaluation and presenta-
tion of data.
ReadyToProcess WAVE 25
Cue Card (29058521)
Brief instructions providing an overview of how to
run the system.
UNICORN Quick Installation
Guide (29414475)
Guide for installation of UNICORN.
UNICORN Administration
and Technical manual
Overview and detailed description of network
setup and complete software installation. Admin-
istration of UNICORN and the UNICORN database.
UNICORN Online Help Dialog descriptions for UNICORN 7.x.
User Documentation CD CD containing the listed manuals and translated
versions of ReadyToProcess WAVE 25 Operating
Instructions.
1 Introduction
1.3 Associated documentation
ReadyToProcess WAVE 25 User Manual 29009598 AD 9

1.4 Abbreviations
Introduction
This section explains abbreviations that appear in the user documentation for this
product.
Concepts and abbreviations used in this manual are explained in the table below.
Concept/
abbreviation
Explanation
Cellbag™ bioreactor The disposable container in which the cells are
cultured.
DO Dissolved oxygen.
DO sensor Optical sensor for measurement of dissolved
oxygen. Attached to DO configured Cellbag
bioreactors.
Single mode Operating mode with one Cellbag bioreactor on the
rocker.
Dual mode Operating mode with two Cellbag bioreactors on
the same rocker. Cultivation is monitored and
controlled independently in the two bioreactors.
pH sensor Optical sensor for pH measurement. Attached to pH
configured Cellbag bioreactors.
ReadyToProcess CBCU Control unit for gas mix, pH and DO control.
ReadyToProcess Pump 25 The pump unit.
ReadyToProcess WAVE 25
rocker
The rocker.
Tray Tray for Cellbag, mounted on the rocker. Different
tray sizes are available for different culture capaci-
ties.
ReadyToProcess WAVE 25
The ReadyToProcess WAVE
25 system
The bioreactor system
The entire bioreactor system, including rocker,
CBCU(s), and pump(s), together with Cellbag
bioreactor(s) and filter heater(s).
UNICORN The software used for controlling and monitoring
the system.
1 Introduction
1.4 Abbreviations
10 ReadyToProcess WAVE 25 User Manual 29009598 AD

2 System description
About this chapter
This chapter gives an overview of ReadyToProcess WAVE 25 and describes the
different bioreactor system units.
In this chapter
Section See page
2.1 System overview 12
2.2 ReadyToProcess WAVE 25 rocker 16
2.3 ReadyToProcess CBCU 24
2.4 ReadyToProcess Pump 25 27
2.5 Cellbag bioreactor 29
2.6 UNICORN software overview 32
2 System description
ReadyToProcess WAVE 25 User Manual 29009598 AD 11

2.1 System overview
Introduction
ReadyToProcess WAVE 25 is intended for cell cultivation.
A disposable Cellbag bioreactor is placed on a rocker and filled with gas, partially filled
with culture medium, and inoculated with cells. Gas transfer and mixing of culture is
accomplished by wave-induced agitation, performed by the rocker unit.
The cell culture volume range per Cellbag bioreactor is 0.3 to 25 L depending on
bioreactor size, and working volume may be expanded up to 10 times during one culti-
vation.
The system, composed of the rocker, ReadyToProcess CBCU and ReadyToProcess
Pump 25 , enables measurement and control of pH, DO, weight and media distribution,
and provides different gas flow and gas mixing possibilities.
•In single mode , the system supports culture in one Cellbag bioreactor at one time.
The rocker is connected to one ReadyToProcess CBCU and up to three
ReadyToProcess Pump 25 units.
•In dual mode , the system supports culture in two Cellbag bioreactors placed on
the same tray. The rocker is connected to up to two ReadyToProcess CBCU units
and up to three ReadyToProcess Pump 25 units for independent control of culture
conditions in the two bioreactors.
The system is controlled from a PC running UNICORN software version 7 or later. The
system can also be controlled from a supervisory control and data acquisition (SCADA)
system, such as the DeltaV™ control system, using the integrated OPC server. Contact
Cytiva for instructions and guidance for OPC.
2 System description
2.1 System overview
12 ReadyToProcess WAVE 25 User Manual 29009598 AD

Illustration of the system
The illustration below shows the main system units for use in single mode with one
ReadyToProcess Pump 25 . Dual mode uses two ReadyToProcess CBCU units for
controlling the two Cellbag bioreactors independently. Both single and dual modes
can support up to three ReadyToProcess Pump 25 units.
1 432
58 7 6
Part Description
1 Hatch
2 Filter heater
3 Cellbag bioreactor
4 ReadyToProcess Pump 25
5 ReadyToProcess CBCU
6 ReadyToProcess WAVE 25 rocker
7 Tray
8 Lid
2 System description
2.1 System overview
ReadyToProcess WAVE 25 User Manual 29009598 AD 13

Illustration of wave motion
1
2
4
3
Stage Description
1 Inflowing gas from the CBCU enters the Cellbag bioreactor through the
inlet vent filter. The gas flow inflates the bag and oxygenates the culture.
2 Metabolic waste gases leave the Cellbag bioreactor through the outlet
vent filter. The pressure control valve maintains a constant overpressure
inside the Cellbag bioreactor.
3 The rocking mechanism sets the rocking platform in motion.
4 Wave motions are induced by the rocking. The culture is cautiously
mixed, and gases are transferred into the culture.
2 System description
2.1 System overview
14 ReadyToProcess WAVE 25 User Manual 29009598 AD

Available controlled parameters
The table below describes the controlled parameters for the fully configured bioreactor
system. The controlled parameters of your configuration may vary from the list below.
In dual mode, all parameters except rocking speed, angle and motion may be
controlled independently in the two Cellbag bioreactors.
Parameter Description
Temperature The system can control temperature between room temperature
plus 5°C and 40°C.
Rocking
speed
Rocking speed can be set between 2 and 40 rpm.
Rocking
angle
Rocking angle can be set between 2 and 12 degrees.
Rocking
motion
Rocking motion can be set between 15% and 100%, and is by
default set to 30%. A rocking motion of 15% gives a uniform
motion with almost constant angular velocity throughout the
movement, and 100% gives a smooth, completely sinusoidal
motion.
Gas flow The system can control the gas flow into the cellbag between 0.02
and 1.00 lpm.
Media
distribution
The system can control culture medium addition and removal.
Two different modes exist, media addition and perfusion.
pH pH is measured and controlled between pH 6 and pH 8. Three
different modes of pH regulation exist, with CO2, CO2/base, and
acid/base.
DO DO is measured between 0% and 250% and controlled between
0% and 100% air saturation. Three different modes of DO regula-
tion exist, with O2, speed, and O2/speed.
CO2CO2 concentration in gas mix can be controlled between 0% and
15% and measured between 0% and 20%.
O2O2 concentration in gas mix can be measured between 0% and
50%, and controlled between 0% and 50% with N2 and between
21% and 50% with air.
2 System description
2.1 System overview
ReadyToProcess WAVE 25 User Manual 29009598 AD 15

2.2 ReadyToProcess WAVE 25 rocker
Introduction
The rocker is the main unit of the system. Through the rocker, weight is measured, and
temperature, rocking speed, rocking angle and rocking motion are controlled.
The rocker contains four load cells for monitoring the weight of the Cellbag bioreactor
and content. The placement of the load cells allows independent weight measurement
of the two Cellbag bioreactors in dual mode.
The rocker also contains an embedded microprocessor, which allows the system to be
controlled independently of the performance of the connected network and client
computer.
For rocker specifications, see Rocker specifications, on page 245.
Rocking parameters
The adjustable rocking parameters are rocking speed, rocking angle and rocking
motion. These factors, in combination with the cell culture volume, have a direct
impact on the oxygen transfer rate and mixing time in the Cellbag bioreactor.
The rocking motion sets how large part of the rocking cycle that has a sinusoidal
angular velocity. It can be adjusted to be more or less sinusoidal. The minimum value,
15%, gives a uniform motion with almost constant angular velocity throughout the
movement, resulting in a step-wise rocking. The maximum value, 100%, gives a sinus-
oidal motion with slower angular velocity in the end positions and faster in the middle
of the movement, resulting in a smoother rocking.
Front view of the rocker
The illustration below shows the front view of the rocker.
1
345
2
2 System description
2.2 ReadyToProcess WAVE 25 rocker
16 ReadyToProcess WAVE 25 User Manual 29009598 AD

Part Description
1 Rocker platform
2 Temperature sensors
3 Rocker base
4Power button
5 Location of adjustable foot
Power button
The Power button indicates the status of the rocker according to the list below.
Light indicator Image Description
No light The power is OFF.
Green flashing light The rocker is starting up.
Green steady light The power is ON and the rocker is
operational.
Red flashing light The rocker failed to connect to other
components in the system.
Red steady light Indicates an error of the rocker.
Adjustable foot
The adjustable foot is placed in the front right corner of the rocker base when viewed
from the front. It is used to distribute weight evenly over the four rocker feet.
Use the supplied adjustable foot wrench to adjust the foot.
2 System description
2.2 ReadyToProcess WAVE 25 rocker
ReadyToProcess WAVE 25 User Manual 29009598 AD 17

Rear view of the rocker
The illustration below shows the rear panel of the rocker.
1 2 4
3
5 6 7
Part Description
1 15-pin D-sub connector, used for digital and analog I/O signals
2 Filter heater connectors
3 Tray connector
4 UniNet-9 ports
5 USB ports
6 Ethernet connector
7 Power connector
Note:
The rocker is fitted with internal electrical fuses that are not user-replace-
able.
Tray and lid sizes
Trays and lids are available in the different sizes listed below:
Trays Lids
Tray 10 Lid 10
Tray 20 Lid 20
Tray 50 Lid 50
2 System description
2.2 ReadyToProcess WAVE 25 rocker
18 ReadyToProcess WAVE 25 User Manual 29009598 AD

Illustrations of tray and lid
The illustration below shows the rocker with Tray 50 attached.
Part Description
1 Bag clamp (upper)
2 Bag clamp opener (one in each upper corner)
3 Bag clamp opener (one in each lower corner)
4 Bag clamp (lower)
The illustration below shows the rocker with Tray 50 and Lid 50 mounted.
2 System description
2.2 ReadyToProcess WAVE 25 rocker
ReadyToProcess WAVE 25 User Manual 29009598 AD 19

54
3
2
1
Part Description
1 Rocker base
2 Lid
3 Tray
4 Tubing exit
5 Hatch
2 System description
2.2 ReadyToProcess WAVE 25 rocker
20 ReadyToProcess WAVE 25 User Manual 29009598 AD